Abstract：ObjectiveThis study was designed to compare inflammatory response, water carriage and gut brain axis in rats with ulcerative colitis (UC) after treatment of three regiments, Huangqintang (HQT), Sishenwan (SSW), and Tongxie Yaofang(TXYF).MethodAfter approved by Institute of Chinese Materia Medica Ethics Committees in China Academy of Chinese Medical Sciences, UC in rats was induced by using a compound method (trinitrobenzenesulfonic acid plus ethanol). Rats were randomly divided into control, disease, positive control salazosulfapyridine (SASP, 0.5 g·kg-1), HQT (20 g·kg-1), SSW(26 g·kg-1), and TXYF group(22 g·kg-1). After 5 days of treatment, colonic tissues and the blood were taken for various assays. Damage of colonic tissues was detected by hematoxylin-eosin staining (HE). The distribution of Vasoactine intrestinal (VIP), 5-hydroxytrytamine (5-HT), P-substance (SP) in the blood and serum were detected by enzymelinked immunosorbent assay (ELISA) and immunohistochemistry (IHC), the levels of aquaporin3 (AQP3) and Aquaporin4 (AQP4) in the serum were detected by Western blot, the mRNA expression of Extracellular regulated protein kinases 1 (Erk1) and p38 mitogen-activated protein kinase (p38 MAPK) in the serum were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR).ResultThe brain-gut peptide results showed that compared with the normal group, the content of 5-HT and VIP in model group were significantly decreased (P<0.01), the content of SP were decreased, but there was no significant statistical difference, compared with the disease group, the content of 5-HT in SASP and TXYF group were clearly increased (P<0.05), the increment of VIP and SP in SASP, HQT, TXYF group were significant (P<0.05). Compared with the normal group, the content of AQP3 in model group were significantly increased(P<0.01), the content of AQP4 were clearly decreased(P<0.01), compared with the disease group, the content of AQP4 in SASP and HQT group were clearly increased (P<0.05), whereas the levels of AQP3 in HQT group were most significant reduced (P<0.01). Compared with the disease group, the expression of Erk1 and p38 were clearly reduced (P<0.01), with the most significant reduce being the expression in HQT group.ConclusionThree regiments all have therapeutic effects on UC, manifested by improvements of the signs and mental status of UC rats. However, in terms of gut-brain axis disturbance improvement, the therapeutic effect of TXYF was superior than HQT and SSW, whereas in terms of inflammatory response suppression and water carriage accomodation, the therapeutic effect of HQT was superior than SSW and TXYF.
Keywords：ulcerative colitis;brain gut peptide;aquaporin;inflammatory response;function mechanism
Abstract：ObjectiveTo study the effect of Biejia Jianwan on expressions of signal molecules and target genes of transforming growth factor-β (TGF-β)/Smad pathway in diethylnitrosamine (DEN)-induced rat hepatocellular carcinoma, and explore the mechanisms of Biejia Jianwan suppressing the invasion of hepatocellular carcinoma.MethodThe rats were divided into three group, namely normal group, model group and Biejia Jianwan group (2.2 g·kg-1·d-1). Rats in Biejia Jianwan group and model group received intraperitoneal injections of DEN to induce sequential chronic inflammation, cirrhosis and hepatocellular carcinoma. At the sign of cirrhosis, rats in Biejia Jianwan group began taking Biejia Jianwan by gavage for 6 weeks. Rat blood was collected to measure serum levels of biochemical markers of liver function tests, including alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bilirubin(TBIL), albumin(Alb), γ-glutamyl transpeptadase(GGT), alkaline phosphatase(ALP). Rat livers were fixed in formalin and stained with hematoxylin-eosin (HE)staining， quantitative real-time PCR was used to test the mRNA expressions of TGF-β1， and Western blot was used to test protein expressions of TGF-β1, Smad2/3, p-Smad2/3, N-cadherin, E-cadherin and Vimentin.ResultAll of the levels of biochemical markers showed no difference in Biejia Jianwan group and model group. Biejia Jianwan could improve the pathological changes of balloon-like degeneration, edema, and necrosis in liver cancer tissues. Importantly, the treatment dramatically decreased the mRNA expression of TGF-β1(P<0.01), and the protein expressions of TGF-β1, p-Smad2(P<0.01). Besides, the protein expression of N-cadherin and Vimentin were decreased significantly (P<0.01).ConclusionBiejia Jianwan can inhibit epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma cells activated via TGF-β/Smad pathway by reducing TGF-β1 expression, so as to suppress the metastasis and invasion of hepatocellular carcinoma.
Abstract：ObjectiveTo study the effect of modified Erchentang on levels of interleukin-12 (IL-12), interferon-γ (IFN-γ), interleukin-9 (IL-9), interleukin-4 (IL-4) and interleukin-13 (IL-13) in plasma and bronchoalveolar lavage fluid (BALF) of all rats, as well as expressions of interleukin-4 (IL-4) receptor (IL-4R1) and interleukin-13 (IL-13) receptor (IL-13RA1) in bronchioles tissue of rats with chronic obstructive pulmonary disease (COPD).MethodFifty SD rats were randomly divided into 5 groups, namely normal group, model group, and low, middle and high-dose modified Erchentang groups (5, 10, 20 g·kg-1), with 10 rats in each group. COPD in rat was prepared by using cigarette smoke combined with dripping lipopolysaccharide (LPS) in trachea. After the modeling, normal and model groups were given normal saline solution through intragastric (ig) administration, while other groups were given corresponding herbal drugs (5, 10, 20 g·kg-1) intragastrically (ig) for 14 days. The levels of IL-12, IFN-γ, IL-9, IL-4 and IL-13 in plasma and BALF were detected by Enzyme-linked immunosorbent assay (ELISA) method， and immunohistochemistry (IHC) method was used to detect the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue of all of the groups.ResultCompared with the normal group, the levels of IL-12 and IFN-γ were decreased significantly (P<0.01), but the levels of IL-9, IL-4 and IL-13 in plasma and BALF were significantly increased (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were increased significantly (P<0.01) in model group. Compared with the model group, the levels of IL-12 and IFN-γ were increased significantly, while the levels of IL-9, IL-4 and IL-13 in plasma and BALF were decreased significantly (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were decreased significantly (P<0.01) in modified Erchentang groups (10, 20 g·kg-1).ConclusionModified Erchentang has effects in resisting inflammatory and protecting tissue structure of bronchioles. Its mechanism may be correlated with increasing the levels of IL-12, IFN-γ and reducing the levels of IL-9, IL-4 and IL-13 in plasma and BALF, and inhibiting the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue.
Abstract：ObjectiveTo explore the intervention effect of modified Shengjiangsan in rats with membranous nephropathy (MN) and its related mechanism.MethodRats were injected with cationized bovine serum Albumin (C-BSA) in the tail vein to establish a rat model of membranous nephropathy. The normal group, model group, modified Shengjiangsan group (27.3 g·kg-1) and benazepril group (10 mg·kg-1) were established in this study. Each group was given corresponding dosage of the drug once a day for 4 weeks of continuous intervention. After the administration, the levels of 24-hour urine protein (UTP), total cholesterol (TC), triglyceride (TG), total protein (TP), Albumin (Alb), creatinine (SCr), urea nitrogen (BUN) level was detected. we observed the pathological changes of rat kidneys by the technology of Masson staining, silver hexylamine iodate (PASM) staining and transmission electron microscopy. immunofluorescence technology was used to detect immunoglobulin （Ig）G deposition in rat kidneys. Western blot was used to detect the expression levels of key proteins in phosphatidylinositol 3-kinase/proline protein kinase B/rapamycin target protein (PI3K/Akt/mTOR) signaling pathway and autophagy marker proteins LC3 and Beclin1.ResultCompared with normal group, the UTP, serum TC and TG levels were significantly increased, TP and Alb levels were significantly reduced in model group(P<0.05). We detected the kidney pathological changes include of glomerulus enlargement, basement membrane thickening,vacuolar degeneration, pheotropin deposition, glomerular capillary loop IgG diffuse deposition, electron dense deposits of varying sizes and podocytes under the epithelium extensive integration of foot processes， the expression of p-PI3K, p-Akt and p-mTOR protein was significantly increased (P<0.05). The expression of autophagy marker proteins LC3 and Beclin1 protein decreased significantly(P<0.05). Compared with model group, the UTP, serum TC and TG levels were decreased in the benazepril group and modified Shengjiangsan group, and the TP and Alb levels were increased (P<0.05)， the histopathological changes of rat kidney were all reduced， the expression of p-PI3K, p-Akt and p-mTOR protein was significantly reduced(P<0.05)， autophagy marker proteins LC3 and Beclin1 protein expression were significantly increased.ConclusionModified Shengjiangsan can reduce urinary protein, reduce kidney pathological damage and delay disease progression, which is related to its inhibition of PI3K/Akt/mTOR signaling pathway and activation of renal autophagy.
Keywords：modified Shengjiangsan;membranous nephropathy;phosphatidylinositol-3kinase/proline protein kinase B/rapamycin target protein (PI3K/Akt/mTOR) signaling pathway;autophagy
Abstract：ObjectiveTo investigate the effect of Huangqi Jianzhongtang on Janusprotein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signal pathway in rats with spleen-stomach deficiency cold type gastric ulcer (GU).MethodA total of 60 SPF level Wistar rats were randomly divided into two groups: blank group and model group. Model rats were used to reconstruct the spleen-stomach deficiency cold type GU model by comprehensive modeling method. Model rats were divided into model group, Anweiyang group and high, medium and low-dose Huangqi Jianzhongtang groups according to the random number table, with 10 rats in each group. Rats in blank group and model group were given 10 mL·kg-1·d-1 distilled water, and 16, 8, 4 g·kg-1·d-1 Huangqi Jianzhongtang, respectively. Rats in the positive control group were given 0.14 g·kg-1·d-1 Anweiyang for 21 days. The gene expressions of JAK2 and STAT3 in the ulcer tissue were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), the protein expressions and phosphorylation levels of JAK2 and STAT3 in the ulcer tissue were detected by Western blot, and the contents of interleukin-10（IL-10）and interleukin-17（IL-17）in the gastric tissue of each group were detected by enzyme-linked immunosorbent assay (ELISA).ResultCompared with the blank group, the general survival condition of the model group was worse, the content of IL-10 in gastric homogenate was significantly reduced, while the content of IL-17 was significantly increased (P<0.05), the protein expressions of JAK2 and STAT3 in gastric tissue was not significantly increased, whereas the gene expressions and phosphorylation levels of JAK2 and STAT3 were significantly increased (P<0.05). Compared with the model group, the content of IL-10 increased, but the content of IL-17 decreased, the gene expressions of JAK2 and STAT3 and the level of protein phosphorylation decreased in the treatment group, especially in the high-dose Huangqi Jianzhongtang group, with statistically significant differences (P<0.05).ConclusionHuangqi Jianzhongtang can improve the survival condition of rats with spleen stomach deficiency cold type gastric ulcer, and its mechanism may be related to the intervention of gastric mucosal immune barrier dysfunction mediated by JAK2/STAT3 pathway activation.
Keywords：Huangqi Jianzhongtang;gastric ulcer;immune barrier;Janusprotein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) pathway
Abstract：ObjectiveTo observe protective effect of Danggui Buxuetang (DBT) on oxidative stress injury of mouse bone marrow-derived endothelial progenitor cells (EPCs) against induced by hydrogen peroxide (H2O2).MethodMonocytes from bone marrow of mice were obtained by density gradient centrifugation, and EPCs were obtained by specific culture medium. The experiment was divided into blank group，model group，DBT group (100，200，400 mg·L-1). Methyl thiazolyl tetrazolium（MTT） assay was used to determine the survival rate of EPCs and establish the cell injury model induced by H2O2. MTT，transwell chamber，matrigel and superoxide fluorescent anion probe (DHE) were used to detect the proliferation，migration，in vitro angiogenesis and ROS level，detection of autophagy by Western blot.ResultCompared with blank group，the proliferation ability，migration ability，the number of lumens and the length of tubule branches of EPCs in the model group were significantly reduced (P<0.01)，the level of intracellular reactive oxygen species （ROS） was significantly increased (P<0.01)，the expression of p62，the light chain microtubule associated protein 1 protein light chain 3 Ⅱ type （LC3-Ⅱ） protein of microtubule associated protein 1，was significantly increased (P<0.01). Compared with model group，DBT group increased the ability of cell proliferation and migration （P<0.01）. In addition，DBT increased the expression of LC3-Ⅱ protein in a concentration dependent manner (P<0.01)，and decreased the expression of p62 protein (P<0.01).ConclusionDBT can improve the autophagy level of EPCs under oxidative stress, promote the proliferation, migration and angiogenesis of injured EPCs, and protect the biological function of EPCs under oxidative stress.
Abstract：ObjectiveTo study the protective effect of Euphorbia helioscopia alcohol extract on lipopolysaccharide (LPS) -induced acute lung injury in mice and explore its possible mechanism.MethodThe 50 Balb/c male mice were randomly divided into 5 groups, including normal group, model group, dexamethasone group (1.5 mg·kg-1), E. helioscopia alcohol extracts group (7.5，3.75 g·kg-1). Except for the normal group, the other groups used intranasal instillation of LPS to establish a model of acute lung injury in mice. The type and number of inflammatory cells in bronchoalveolar lavage fluid (BALF) were detected by automatic blood analyzer and Wright-Giemsa composite staining. The lung tissue damage was observed by hematoxylin-eosin (HE) staining. The contents of the inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in BALF were detected by flow cytometry. The protein expressions of nuclear factor kappa-B p65(NF-κB p65), phospho-NF-κB p65 (p-NF-κB p65), inhibitor of NF-κBα (IκBα), phospho-IκBα (p-IκBα) in NF-κB pathway and c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), p38 protein (p38), phospho-p38 (p-p38), extracellular regulated protein kinases (ERK1/2), phospho-ERK1/2 (p-ERK1/2) in mitogen-activated protein kinase (MAPK) pathway were determined by Western blot.ResultCompared with normal control group, the lung tissue of the model group showed obvious damage, in which a large number of inflammatory cells infiltrated, and the integrity of the alveoli was destroyed. Inflammatory factors TNF-α, IL-6 in BALF and p-NF-κB p65, p-JNK, p-p38, p-ERK protein expression levels in lung tissue were significantly increased (P<0.01). Compared with model group, the pathological damage of lung tissue in mice with high dose of E. helioscopia alcohol extract and dexamethasone positive group was significantly alleviated. The levels of TNF-α and IL-6 in BALF and the expression levels of p-NF-κB p65, p-JNK, p-p38 and p-ERK1/2 protein in lung tissue were significantly down-regulated (P<0.01).ConclusionThe E. helioscopia alcohol extract has a protective effect on LPS-induced acute lung injury in mice, its mechanism may be related to the regulation of the NF-κB/MAPK signaling pathway.
Abstract：ObjectiveTo observe Plantaginis Semen's mechanism in treating diarrhea by observing the effect on inflammatory factors in serum and mRNA and protein expressions of aquaporin4 (AQP4) in colon tissue of diarrhea rats.MethodSenne Folium was orally administered to duplicate diarrhea rats. Sixty male SD rats were randomly divided into normal group, model group, hydrochlorothiazide group (9 mg·kg-1), and low, middle, and high-dose Plantaginis Semen groups (0.95, 1.9, 3.8 g·kg-1). Senne Folium (20 mL·kg-1) was intragastrically administered in 5 groups in the morning, except for normal group that was orally given the same dose of distilled water. In the afternoon, each treatment group was orally given the corresponding drugs, while normal group and model group were orally given the same dose of distilled water. The loose stool rate, average degree of loose stool, and diarrhea index were compared according to fecal traits and stool times after 14 days of treatment. The serum and colon tissue were collected to detect the contents of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and C-reactive protein (CRP) in serum. Hematoxylin-eosin (HE) staining was used to observe the pathological morphological changes of colon tissue, and quantiative Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expressions of AQP4 in colon tissue.ResultIn the model group, the loose stool rate, average degree of loose stool, and diarrhea index were significantly increased (P<0.01), apoptosis and necrosis were observed in the epidermal cells of colonic mucosa, telangiectasia and congestion in lamina propria were obvious, and a few neutrophils were infiltrated, and the contents of TNF-α, IL-6 and CRP in serum increased (P<0.05, P<0.01), the mRNA and protein expressions of AQP4 significantly decreased (P<0.01). Compared with the model group, the loose stool rate, average degree of loose stool, and diarrhea index were significantly decreased in low, middle, and high-dose Plantaginis Semen groups (P<0.01), the apoptosis and necrosis of epidermal cells, telangiectasia and hyperemia and neutrophil infiltration in colonic mucosa were obviously improved, and the contents of TNF-α and CRP in serum significantly decreased (P<0.05, P<0.01), the mRNA and protein expressions of AQP4 increased （P<0.05, P<0.01）.ConclusionPlantaginis Semen has a better antidiarrheal effect, and its mechanism may be related to inhibition of inflammatory reaction, repair of pathological damage of colonic mucosa, up-regulation of AQP4 expression and promotion of water and fluid metabolism.
Keywords：diarrhea;Plantaginis Semen;tumor necrosis factor-α (TNF-α);interleukin-6 (IL-6);C-reactive protein (CRP)
Abstract：ObjectiveAstragalus polysaccharide (APS) was used in combination with ionizing radiation (IR) to investigate the mechanism of APS on the radiosensitivity of human nasopharyngeal narcinoma CNE-1 cells and the epithelial-mesenchymal transition (EMT).MethodCell counting kit-8 (CCK-8) was used to detect the cytotoxicity of different concentrations of APS (0，6.25，12.5，25，50，100，200 g·L-1) on CNE-1 cells. Colony formation assay was used to calculate the survival fraction (survival fraction, SF) of CNE-1 cells treated with 12.5 g·L-1 APS combined with different radiation doses (0，2，4，6 Gy). The linear quadratic equation mathematical model (LQ) was used to draw the radiosensitivity curve according to SF value. Cell scratch and transwell chamber test were used to detect the migration and invasion ability of cells in each group. The apoptosis of cells in each group was detected by flow cytometry, Western blot was used to detect the expressions of EMT markers, apoptosis markers and protein kinase B/extracellular regulated protein kinases (Akt/ERK) pathway proteins in each group.ResultThe results of colony formation assay and radiosensitivity curve showed that the combination of non-toxic dose of 12.5 g·L-1 APS and radiation dose of 4 Gy could significantly increase the radiosensitivity of CNE-1 cells. Compared with blank group and IR group, APS combined with IR could significantly inhibit the migration and invasion of CNE-1 cells (P<0.05), and increase the rate of apoptosis (P<0.05). In addition, compared with the blank group and the IR group, APS combined with IR could significantly down-regulate the expressions of N-cadherin, p-Akt and p-ERK, and significantly up-regulate the expressions of E-cadherin, Bax and Caspase-3 (P<0.05).ConclusionAPS combined with IR can inhibit the migration and invasion of CNE-1 cells, and increase the apoptosis induced by radiotherapy, which may be related to the inhibition of EMT and Akt/ERK pathway.
Abstract：ObjectiveTo investigate the therapeutic effect of Qihong capsule on pentobarbital sodium induced heart failure in beagle dogs.MethodThirty healthy adult beagle dogs were randomly divided into 6 groups, 6 in each group. They were normal group, model group, digoxin tablet group (40 μg·kg-1), Qihong capsule high, medium and low dose groups (2.6，1.3，0.65 g·kg-1). The heart failure model of beagle dogs was established by intravenous infusion of 2% pentobarbital sodium. The success standard of the model was that the maximum rate of rise of left ventricular pressure was reduced by 70%.The corresponding drugs were given through duodenum. The Ⅱ lead electrocardiogram, coronary blood flow, cardiac output, left ventricular pressure and maximum rate of left ventricular pressure rise were measured by multi-channel physiological recorder. The arterial oxygen content and coronary sinus oxygen content were measured by Roche blood oxygen analyzer at different time points, and the myocardial oxygen utilization rate was calculated.ResultAfter intravenous infusion of 2% pentobarbital sodium for about 15 minutes, beagle dogs began to show obvious symptoms of heart failure. The main manifestations were the increase of PR interval of Ⅱ lead electrocardiogram, the decrease of coronary blood flow, left ventricular pressure, cardiac output, cardiac output, venous oxygen content, and the increase of myocardial oxygen utilization rate (P<0.01) compared with the model group, Qihong capsule significantly increased coronary blood flow at 60-120 min after treatment (P<0.05). The cardiac output of 2.6 g·kg-1 Qihong capsule increased significantly at 45-60 min after treatment, with the maximum increase of about 16%, which was significantly different from that of model group (P<0.05). At the same time, it can increase the oxygen content of coronary sinus blood, which indicates that the myocardial oxygen supply is increased and the oxygen utilization rate is decreased. Qihong capsule 1.3 g·kg-1 group significantly increased the maximum rate of left ventricular pressure rise (P<0.05), the maximum increase rate was about 42%.ConclusionQihong capsule can increase coronary blood flow and venous blood oxygen content at the same time, make myocardial nutrient supply sufficient, reduce oxygen utilization rate, on this basis, Qihong capsule can further increase cardiac output and improve cardiac function, so as to play a protective role in heart failure.
Keywords：heart failure;hemodynamics;Qihong capsule;coronary blood flow;myocardial consumption of oxygen
Abstract：ObjectiveTo observe the effect of blood stasis syndrome on renal damage and endoplasmic reticulum stress (ERS) in diabetic kidney disease (DKD) rats, and to explore the relationship between renal syndrome of blood stasis damage and ERS in DKD rats.MethodThe 50 Male SD rats of SPF level were selected to establish DKD rat model by high fat and high sugar diet combined with intra-abdominal injection of streptozotocin(STZ). They were randomly divided into normal group, diabetes mellitus group and diabetes mellitus and blood stasis syndrome group（0.05 mg·kg-1）, among which diabetes mellitus and blood stasis syndrome group was prepared by dextran method. The general conditions, hemorheology indexes, 24 h urine protein, serum creatinine and renal pathology of the rats in each group were observed. Immunohistochemical analysis, Western blot and Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were used to detect mRNA and protein expressions of endoplasmic reticulum stress-related proteins glucose regulated protein 78(GRP78), activating transcription factor-6(ATF6) and renal fibrosis index fibronectin(FN), and alpha smooth muscle actin(α-SMA).ResultCompared with normal group, the rats in diabetes mellitus group showed polyphagia, polyuria and weight loss, increased hemorheology index of whole blood viscosity and plasma viscosity(P<0.05，P<0.01), increased 24 h urine protein, serum creatinine and urea nitrogen(P<0.01), and increased renal pathology, and increased mRNA and protein expression of GRP78, ATF6, FN and α-SMA(P<0.05，P<0.01). After dextran preparation of blood stasis model. Diabetes mellitus and blood stasis syndrome group increased mortality, signs of change is more obvious in the diabetic group, whole blood viscosity, plasma viscosity, 24 h urine protein ration, serum creatinine and urea nitrogen were significantly higher than those in diabetic rats(P<0.01), pathological changes aggravated in the diabetes group. At the same time， mRNA and protein expressions of GRP78, ATF6, FN, and α-SMA in renal tissue were significantly higher than those in diabetic mellitus group(P<0.05，P<0.01).ConclusionUnder the combined disease and syndrome model, the blood stasis syndrome may further aggravate the pathological damage of the kidney of DKD rats, and is related to the enhancement of ERS in the kidney of DKD rats.
Keywords：diabetic kidney disease(DKD);syndrome of blood stasis;endoplasmic reticulum stress(ERS);ombination of disease and syndrome
Abstract：ObjectiveTo observe the effect of Puerariae Lobatae Radix on type 2 diabetes mellitus (T2DM) rats and related mechanism.MethodNinety SD rats were divided into normal group, model group, metformin group, Puerariae Lobatae Radix high dose group, Puerariae Lobatae Radix medium dose group and Puerariae Lobatae Radix low dose group, 15 rats in each group. The rats in abnormal group were fed with high-fat and high sugar diet for 4 weeks, and then T2DM model was established by intraperitoneal injection of 40 mg·kg-1 streptozotocin (STZ). Puerariae Lobatae Radix high-dose group was intragastrically administered with 2.1 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix medium-dose group was intragastrically administered with 1.4 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix low-dose group was intragastrically administered with 0.7 g·kg-1 of Puerariae Lobatae Radix extract powder, 0.2 g·kg-1 of metformin hydrochloride in metformin group, distilled water once a day in normal group and model group. After 8 weeks, fasting blood glucose (FBG), glycosylated serum protein (GSP), insulin (FINS), triglyceride (TG), cholesterol (TC) were measured, and insulin resistance index （HOMA-IR） was calculated. The expression of tumor necrosis factor -α(TNF-α) was observed by immunohistochemistry. Western blot was used to detect the protein expression of glucose regulatory protein 78 （GRP78）, activated transcription factor 6 （ATF6） in pancreatic tissue.ResultCompared with normal group, the contents of FBG, GSP, TG, TC, FINS, TNF-α in model group were significantly increased (P<0.05), the HOMA-IR was significantly increased (P<0.05), the protein expressions of GRP78 and ATF6 in pancreatic tissue were significantly increased (P<0.05). Compared with model group, the contents of FBG, GSP, TG, TC, FINS and TNF-α in the metformin group and Puerariae Lobatae Radix high, medium and low dose groups were significantly decreased (P<0.05), HOMA-IR decreased significantly (P<0.05), and the expression of GRP78 and ATF6 protein in pancreatic tissue decreased significantly (P<0.05).ConclusionPuerariae Lobatae Radix can significantly improve insulin resistance in T2DM rats, inhibit the expression of inflammatory factor TNF-α in pancreatic tissue, reduce the protein expression of GRP78 and ATF6 in pancreatic tissue.
Abstract：ObjectiveTo clarify the effect of Fangfeng Tongshengtang on early-stage serum endotoxin (ET) and programmed death-1/programmed ligand-1(PD-1/PD-L1) in patients with hepatitis B virus-related acute-on-chronic liver failure（HBV-ACLF）(early-stage), and exploring the mechanism of Fangfeng Tongshengtang in the treatment of early stage HBV-ACLF.MethodThe 69 patients with early stage HBV-ACLF were enrolled in the study and all of them received antiviral drugs, liver protection and jaundice relieving drugs as well as supporting therapy. According to the random number table, 35 patients were randomly assigned to observation group (to take Fangfeng Tongshengtang, and 34 patients were assigned to control group to take placebo. The observation period was 3 weeks in both groups. Before treatment and 1, 2, and 3 weeks after treatment, theserum ET, expression of PD-1/PD-L1 in serum CD4+ and CD8+ T cells, liver function [alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), total bilirubin (TBIL), and direct bilirubin (DBIL)], coagulation function [prothrombin time (PT), and prothrombin activity (PTA)] were detected to verify the effect of Fangfeng Tongshengtang on HBV-ACLF (early-stage).ResultAfter 3 weeks of treatment, ET, expression of serum CD4+PD-1+, CD4+PD-L1+, CD8+PD-1+, CD8+PD-L1+, ALT, AST, TBIL, DBIL, and PT decreased significantly (P<0.01), while Alb and PTA increased significantly（P<0.01）in both groups. As compared with the control group, the ET in observation group was lower at 1st, 2nd and 3rd week after treatment (P<0.01), the CD4+PD-1+, CD4+PD-L1+, CD8+PD-1+ and CD8+PD-L1+ in observation group were lower at 2nd week and 3rd week（P<0.05, P<0.01）， the ALT, AST, TBIL and DBIL in observation group were lower at 1st, 2nd and 3rd week（P<0.05, P<0.01）， the PT in observation group was lower at 2nd and 3rd week（P<0.05）， and the PTA in observation group was higher at the 2nd and 3rd week（P<0.01）.ConclusionFangfeng Tongshengtang can achieve the therapeutic effect for HBV-ACLF (early-stage) probably by reducing the serum ET and the expression of PD-1 / PD-L1 in serum CD4 +, CD8 + T cells.
Abstract：ObjectiveTo observe the clinical efficacy of Wenjing Huayu Zhitong therapy in treating primary dysmenorrhea (PD) with cold coagulation and blood stasis, and to explore its immune mechanisms on PD.MethodThe 108 PD patients with cold coagulation and blood stasis syndrome were collected and randomly divided into traditional Chinese medicine (TCM) group, ibuprofen group and placebo group according to the random number table method, with 36 cases in each group. All patients received corresponding medicines three days before menstruation. The patients in TCM group were treated with TCM and ibuprofen sustained release capsule simulator. The patients in ibuprofen group were treated with ibuprofen sustained-release capsule and TCM simulator. The patients in placebo group were treated with TCM simulator and ibuprofen sustained-release capsule simulator. Treatment lasted for 6 consecutive days for three menstrual cycles, and follow-up was conducted for three menstrual cycles after drug withdrawal. The visual analogue score (VAS), total time of abdominal pain and TCM symptom scores in each menstrual cycle were recorded. The levels of CD3+, CD4+, CD8+ and the ratio of CD4+/CD8+ in peripheral blood before and after treatment were detected by flow cytometry.ResultAfter treatment for three menstrual cycles, both the TCM group and ibuprofen group were better than placebo group in reducing VAS score and reducing total abdominal pain time (P<0.01). The long-term follow-up effect after drug withdrawal in TCM group was significantly better than that in ibuprofen group (P<0.01). The total effective rate was 91.43% (32/35) in TCM group, 66.67% (10/33) in ibuprofen groups, and 30.30% (10/33) in placebo group . The efficacy of the TCM group was better than that of the ibuprofen group (χ2=-2.971, P<0.01), and the efficacy of the ibuprofen group was better than that of the placebo group (χ2=-2.371, P<0.05). After treatment, the levels of CD3+, CD4+ and CD4+/CD8+ in TCM group were significantly increased and the levels of CD8+ were decreased significantly as compared with those before treatment (P<0.01). After treatment, the levels of CD4+ and CD4+/CD8+ in TCM group were higher (P<0.05，P<0.01)，while the levels of CD8+ were significantly lower than those in ibuprofen group and placebo group (P<0.01).ConclusionWenjing Huayu Zhitong therapy can reduce the VAS score and accumulative time of abdominal pain, and improve the dysmenorrhea symptoms in patients with PD. Reversal of the T cell subsets disorder may be one of its mechanisms in treating PD with cold coagulation and blood stasis.
Abstract：ObjectiveTo investigate the clinical efficacy of Shashen Maidongtang plus total glucosides of paeony capsule on primary Sjogren's syndrome (pSS) based on the theory of fluid metabolism.MethodIn this study, 84 patients of Qi-Yin deficiency type pSS admitted in Zhengzhou Chinese Medicine Hospital from January 2018 to January 2019 were divided into observation group (42 cases) and control group (42 cases) on the basis of random number table. The control group was orally given total glucosides of paeony capsule and iguratimod tablet, while the observation group was orally given Shashen Maidongtang combined with glucosides of paeony capsule. After 3 months of continuous treatment to all subjects, the clinical efficacy was evaluated, and side effects were recorded. Before and after the treatment, the saliva flow rate and basal tear secretion Schirmer I test (SIt) value were measured, European League Against Rheumatism Sjogren's syndrome patient reported index and Sjogren's syndrome disease activity index (ESSPRI and ESSDAI) were scored, the erythrocyte sedimentation rate (ESR) was determined by Westergren, and the levels of serum rheumatoid factor (RF) and immunoglobulin (Ig) G were tested by immunoturbidimetry and rate scattering turbidimetry, respectively.ResultThe overall effective rate of the observation group was 90.48% (38/42), which was much higher than 69.05% (29/42) of the control group (χ2=5.974，P<0.05). After treatment, the saliva flow rates and SIt values of both groups got significantly increased compared with those before the treatment (P<0.05), but the saliva flow and SIt of the observation group were significantly better than those of the control group over the same period after treatment (P<0.05). After treatment, both groups had a great decrease in ESSPRI and ESSDAI scores compared with those before the treatment (P<0.05), and the above scores of the observation group were dramatically lower than those of the control group over the same period (P<0.05). After treatment, ESR, serum RF, and IgG levels of both groups were significantly lower than those before the treatment (P<0.05), and the observation group showed higher levels of ESR, serum RF and IgG than the control group over the same period after treatment (P<0.05). Side effects were few and mild in both groups.ConclusionIn treating patients of Qi-Yin deficiency type pSS, Shashen Maidongtang plus total glucosides of paeony capsule was proven to be effective generally. It could significantly inhibit excessive inflammation and hyperhumoral immunity in patients, and control their disease activity. This may be related to the effect of Shashen Maidong decoction and its decomposed recipes in correcting body fluid infusion and metabolic disorder in patients of Qi-Yin deficiency type pSS.
Keywords：fluid metabolism theory;Shashen Maidongtang;total glucosides of paeony capsule;primary Sjogren's syndrome
Abstract：ObjectiveTo evaluate the short-term and long-term efficacy of Jingui Shenqiwan combined with three-stage syndrome differentiation on osteoporotic vertebral compression fractures (OVCFs) after operation, and its regulatory effect on biomarkers of bone metabolism.MethodOne hundred and thirty-five patients were randomly divided into control group (67 cases) and observation group (68 cases) by random number table. A total of 58 patients in control group completed the treatment (4 patients were exfoliated, 2 patients were lost to follow-up, 3 patients were eliminated); and 60 patients in observation group completed the treatment (3 patients were exfoliated, 2 patients were lost to follow-up, 3 patients were eliminated). Both groups patients were given calcitonin injection through intramuscular injection, 20 u/time, 1 time/week, for 12 weeks, calcium carbonate D3 chewable tablets (Ⅱ), 1 tablet/time, 2 times/day, and alendronate sodium tablets, 70 mg/time, 1 time/week. Patients in control group got Bushen Jiangu capsule, 4 grains/time, 3 times/day. And patients in observation group got modified Jingui Shenqiwan combined with fracture three-stage symptom differentiation, 1 dose/day. The courses of treatment in the two groups were 24 weeks, and a 24 week follow-up was provided. Before the operation and at the 12th and 24th week after operation, the short-term efficacy indexes, such as back pain, lumbar function, traditional Chinese medicine （TCM） syndromes and complications, were recorded. And the long-term efficacy indexes, such as recovery of responsible vertebral body, lumbar function, bone density and quality of life and incidence of 48 week re-fracture, were also recorded. Before and after operation, levels of bone alkaline phosphatase (BALP), osteocalcin (BGP), tartrate resistant acid phosphatase-5b (TRAP-5b), type I collagen carboxy terminal prepeptide (PICP), type I collagen cross-linked C-terminal peptide (β-CTX) and N-MID-OT were detected, and the safety was evaluated.ResultThe comprehensive efficacy in observation group was superior to that in control group (Z=2.026, P<0.05). At the 12th and 24th week after operation, scores of back pain, lumbar function and TCM syndromes were all lower than those in control group (P<0.01), and score of lumbar function at the 48th week after operation was also lower than that in control group (P<0.01). Bone density was higher than that in control group at the 24th and 48th week after operation, and score of quality of life was lower than that in control group (P<0.01). At the 24th and 48th week after operation, Cobb angles were less than those in control group, and heights of responsible centrums (anterior, central, posterior) were higher than those in control group. Cumulative incidence of complications in control group was 51.72% (30/58), which was higher than 26.67% (16/60) in control group (=7.784, P<0.01). The levels of BGP were higher than those in observation group at the 24th and 48th week after operation, and the levels of BALP, TRAP-5b, PICP, β-CTX and N-MID-OT were all lower than those in control group (P<0.01). And there was no side effect relating to Jingui Shenqiwan.ConclusionModified Jingui Shenqiwan combined with fracture three-stage symptom differentiation can reduce the symptoms of back pain and promote the recovery of lumbar function, with a significant short-term comprehensive efficacy. In the long term, it can improve the strength of responsible centrums, restore the anatomical structure of injured centrums, increase the bone density of centrums, further improve the lumbar function, reduce the occurrence of complications, regulate the markers of bone metabolism, and improve osteoporosis.
Abstract：ObjectiveTo discuss clinical effect of comprehensive Qingre Tiaoxue decoction combined with Wuwei Xiaodu drink and enema to sequelae of pelvic inflammatory disease (syndrome of dampness heat stasis) and to study the moderating effect to immune inflammatory factor.MethodOne hundred and sixty patients were randomly divided into control group (80 cases) and observation group (80 cases) by random number table. The 73 patients in control group completed the therapy (4 patients were exfoliated or lost to follow-up, 3 patients were eliminate), 74 patients in observation group completed the therapy (4 patients were exfoliated or lost to follow-up, 2 patients were eliminate). Patients in control group got Fuke Qianjin capsules,2 grains/time, 3 times/day, and at the third day after menstruation, addition and subtraction therapy of Wuwei Xiaodu drink with enema for 14 days at every night, 1 dose/day, and enema continued for 2-4 h/day. Based on the treatment of enema in control group, patients in observation group added Qingre Tiaoxue decoction for 3 menstrual cycles, 1 dose/day, and stopping during menstrual cycle. Degree of pelvic pain, dysmenorrhea and the degree of pain in the lower abdomen and lumbosacral in the non menstrual period were evaluated by pain visual simulation score (VAS). Before and after treatment, scores of syndrome of dampness heat stasis, Mc Cormack scale, and Summary of the World Health Organization Quality of Life Measurement Scale (WHOQOL-BREF) scale were graded, and vaginal ultrasonography, culture of cervical secretion and routine examination of leucorrhea were detected. And levels of CD4+, CD8+, CD4+/CD8+, T lymphocyte subsets, tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2) and IL-6 were detected and safety was evaluated.ResultBy ANOVA of repeated measurement, after treatment, scores of VAS during dysmenorrheal and VAS during non menstrual period in two groups decreased (P<0.05), and during 3 menstrual cycles, scores of VAS during dysmenorrheal and VAS during non menstrual period in observation group were lower than those in control group (P<0.01). Scores of physical sign and syndrome of damp heat and stasis were lower than those in control group (P<0.01), score of WHOQOL-BREF was higher than that in control group (P<0.01). The depth of pelvic effusion and the volume of pelvic inflammatory mass in observation group were less than those in control group (P<0.01). Levels of CD4+, CD4+/CD8+ and IL-2 were higher than those in control group (P<0.01), and levels of CD8+, TNF-α and IL-6 were lower than those in control group (P<0.01). Comprehensive curative effect in observation group was better than that in control group (Z=2.028, P<0.05). And curative effect of traditional Chinese medicine（TCM） Syndrome in observation group was better than that in in control group (Z=2.064, P<0.05). And there were no serious adverse events and adverse reaction caused by Chinese medicine.ConclusionComprehensive therapy of Chinese medicine and enema to sequelae of pelvic inflammatory disease can improve the clinical symptoms and signs, improve the quality of life of patients, and regulate the cellular immune function and inflammatory factors. It has better comprehensive curative effect and TCM syndrome curative effect, and is safe for clinical use.
Keywords：sequelae of pelvic inflammatory disease;syndrome of damp heat and stasis;Qingre Tiaoxiang decoction;Wuwei Xiaodu drink;T lymphocyte subsets;inflammatory factors
Abstract：ObjectiveTo study the effect of combined decoction and single decoction of Houpu Wenzhongtang on rats with deficiency-cold of spleen and stomach from the perspective of metabonomics, to find out the relevant potential biomarkers and metabolic pathways, and to explore the similarities and differences between the combined decoction and single decoction, so as to provide reference for the feasibility analysis of replacing traditional decoction with single dispensing granule of this formula.MethodSD rats were randomly divided into normal group, model group, Houpu Wenzhongtang combined decoction group and simgle decoction group. Rats in the normal group were given distilled water by intragastric administration, rats in the other three groups were given cold vinegar at 4 ℃ in the morning and refined lard in the afternoon for 10 days (the dosage of 10 mL·kg-1). After the model was successfully established, rats in the combined decoction group and the single decoction group were given corresponding decoction with dosage of 1.8 g·kg-1 (according to the amount of crude drugs), once a day for 7 days. Ultra-high liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) technique was used to analyze the small molecular endogenous metabolites in urine. Orthogonal partial least squares-discriminant analysis (OPLS-DA) and principal component analysis (PCA) were used to compare the changes of differential metabolites among the normal group, model group, Houpu Wenzhongtang combined decoction group and single decoction group, and the differential metabolites were introduced into Kyoto Encyclopedia of Genes and Genomes (KEGG) for metabolic pathway analysis.ResultCompared with the model group, the Houpu Wenzhongtang combined decoction group and single decoction group jointly regulated 13 potential biomarkers, including phosphatidylcholine(PC), lysophosphatidic acid(LysoPA) and cholic acid, etc. They played a role in treating deficiency-cold of spleen and stomach by influencing metabolic pathways such as glycerophospholipid metabolism, glycerolipid metabolism, phosphatidylinositol signaling system and so on. The combined decoction and single decoction of Houpu Wenzhongtang could obviously restore the body weight, motilin and gastrin contents of rats with deficiency-cold of spleen and stomach to normal levels.ConclusionAccording to biochemical indexes, there is no obvious difference between combined decoction and single decoction of Houpu Wenzhongtang, but according to metabonomics, the combined decoction may be slightly better than the single decoction. The research shows that it is feasible to replace traditional decoction with single dispensing granule of Houpu Wenzhongtang in clinical application.
Keywords：Houpu Wenzhongtang;combined decoction;simple decoction;metabonomics;biomarkers;deficiency-cold of spleen and stomach;dispensing granules
Abstract：ObjectiveTo optimize the processing technology of salt-processed products of Plantaginis Semen with the specific process parameters, and verify the obtained processing technology by pharmacodynamic research, so as to provide experimental basis for the standardized production and quality control of this decoction pieces.MethodTaking composite score of appearance character score, dry extract yield and contents of three components (geniposidic acid, acteoside and isoacteoside) as index, the analytic hierarchy process (AHP)-criteria importance through intercrieria correlation (CRITIC) mixed weighting method was used to determine the weight coefficient of each index. Based on single factor tests, the response surface method was used to investigate the effects of frying time, frying temperature, salt amount and water amount on the processing technology of salt-processed products of Plantaginis Semen, and the processing technology was verified by diuretic experiment with furosemide tablets as the positive drug (administration dose of 0.01 g·kg-1).ResultThe weight coefficients of geniposidic acid content, acteoside content, appearance character score, isoacteoside content and dry extract yield were 0.319, 0.193, 0.207, 0.273 and 0.008, respectively. The optimal process parameters were as following：fried at 150-180 ℃ for 10 min (obtained from the single factor tests), 100 g of Plantaginis Semen sprayed evenly with 2 g of salt (2 g of salt dissolved in 20 mL of water), and fried at 150-180 ℃ for 15 min. Compared with the blank group, both of the raw products group and the salt-processed products group could significantly increase the secretion of urine volume (P<0.01), but the excretion of Na+ in the urine of rats in the salt-processed products group was significantly higher than that in the raw products group (P<0.05).ConclusionThe optimized processing technology is simple and feasible, which can provide reference for standardizing the industrial production of salt-processed products of Plantaginis Semen. At the same time, combined with inherent quality and appearance of the salt-processed products, and verified by pharmacodynamic test, the obtained results are reasonable and reliable, which can be used for quality control of this decoction pieces.
Keywords：salt-processed products of Plantaginis Semen;processing technology;analytic hierarchy process (AHP);criteria importance through intercrieria correlation (CRITIC);response surface method;pharmacodynamics;diuresis
Abstract：ObjectiveTo construct a systematic identification system of Anemonis Flaccidae Rhizoma, and to evaluate the comprehensive quality of Anemonis Flaccidae Rhizoma from 16 regions in China, so as to lay a foundation for its origin selection and clinical medication safety.MethodThe authenticity of Anemonis Flaccidae Rhizoma was quickly identified by traditional identification method and DNA barcode molecular identification technology, and HPLC-UV was used to determine the contents of 5 active ingredients in Anemonis Flaccidae Rhizoma. All high pressure chromatographic separations were performed with a Welch Ultimate XB-C18 column (4.6 mm×250 mm, 5 μm), the mobile phase consisted of acetonitrile-0.01% trifluoroacetic acid aqueous solution (30∶70) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 210 nm and the column temperature was maintained at 30 ℃.ResultThe authenticity of Anemonis Flaccidae Rhizoma could be precisely and rapidly identified by ribosomal DNA internal transcribed spacer 2 (ITS2) sequence and traditional identification methods. BLAST comparative analysis found that medicinal materials from 16 areas were all Anemone flaccida. Based on the contents of multi-index components, it was shown that the total content of 5 triterpenoid saponins in Anemonis Flaccidae Rhizoma from Banqiao, Enshi, Hubei was the highest (10.59%), followed by Hezhang, Bijie, Guizhou (6.28%) and Duzhenwan, Changyang, Hubei (5.64%).ConclusionDNA barcoding can be used as an effective supplement to the traditional identification technology, it can ensure the authenticity of Anemonis Flaccidae Rhizoma and the safety of clinical use. The comprehensive evaluation of multi-index components of HPLC and cluster analysis show that the quality of medicinal materials in Enshi, Changyang, Wufeng of Hubei, Bijie of Guizhou and Jinfoshan of Chongqing is superior, which can be considered as important origin of Anemonis Flaccidae Rhizoma.
Abstract：ObjectiveTo study the effect of different shading conditions on the growth and photosynthetic physiological parameters of Acanthopanax senticosus，and provide a theoretical basis for standardized planting and rational development and utilization of A. senticosus.MethodThree-year-old A. senticosus was used as the experimental sample.The growth parameters and photosynthetic physiological parameters of plant height，leaf number and leaf area were determined to study the effects of different shading conditions on the growth and photosynthesis of A. senticosus.ResultPlant height，leaf number，and leaf area were significantly higher in the shading treatment than in the control group，and highest under moderate shading conditions.The net photosynthetic rate，stomatal conductance，and transpiration rate were significantly higher in the moderate shading group than in the control group，and decreased in the severe shading group，while the intercellular carbon dioxide concentration was significantly lower in the moderate shading group than in the other treatment groups.As the treatment time progressed，the initial fluorescence was not significantly higher in the moderate shading group than in the control group，but significantly higher in the severe shading group；the maximum fluorescence was significantly higher in the shading group than in the control group，in the moderate shading group.The potential photochemical efficiency and maximum photochemical efficiency were not significantly different between the moderate shading group and the control group and decreased in the severe shading group，which was significantly lower than other treatment groups.ConclusionShading treatment is beneficial to the growth of A. senticosus.The moderate shading condition can significantly improve the photosynthesis of A. senticosus.Severe shading treatment can significantly inhibit the photosynthesis of A. senticosus leaves.This effect is related to the regulation of stomatal regulation. The activity of the photoreaction center is related.
Abstract：ObjectiveTo identify WRKY genes from the transcriptome dataset of Prunella vulgaris by bioinformatics method, and analyze the protein characteristics and expression level of these genes.MethodWRKY transcription factor were identified from the P. vulgaris transcriptome database，their motif，physical and chemical properties，functional annotations，family evolution and expression patterns were analyzed, and their functions were predicted.ResultA total of 23 WRKY transcription factors were identified from P. vulgaris in this study by computational prediction method.Structural analysis found that WRKY proteins contained a highly conserved motif WRKYGQK. Phylogenetic analysis of WRKYs together with the homologous genes from Arabidopsis thaliana could be divided into two groups（group Ⅰ-Ⅱ）. There were 7 members in group Ⅰ，and 16 members in group Ⅱ, group Ⅱ was subdivided into five subgroups，namely group Ⅱb （3 members），Ⅱc（5 members），Ⅱd（3 members），Ⅱe（5 members）. The physical and chemical properties of WRKY protein showed that the amino acid number was between 85 and 599，the molecular weight was between 9 527.5-66 438.45 Da，the theoretical isoelectric point was between 5.01-9.83.Among them, c13719.graph_c0，c32199.graph_c0，c24547.graph_c0，c37881.graph_c0 may play a role in the regulation of secondary metabolitessynthesis of P. vulgaris.And c32199.graph_c0，c26537.graph_c0，c23728.graph_c0 may has an effect in identifying and defensing pathogens in P. vulgaris.The transcriptional profiles of these 23 WRKY genes in various tissues were investigated using transcriptome dataset.The results showed that the expression level of WRKY genes varied significantly in different tissues.ConclusionThis study identifies the organization and transcriptional profiles of PmWRKY genes for the first time, so as to provide the helpful information for further studies of functions of WRKYs.
Abstract：ObjectiveTo study the effect of Qiwei Baizhusan (QWBZS) on liver insulin phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt） signal pathway of diabetic mice induced by high-fat diet and streptozotocin (STZ).MethodThe methods of network pharmacology and animal experiments were used to study the hypoglycemic effect of QWBZS. Active chemical components of the drug and disease targets selected through public databases were used to construct the protein-protein interaction network (PPIN), and gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomics(KEGG) enrichment analysis was performed to identify relevant signal pathways in vivo. In the pharmacological experiment, the diabetic mice model was established through intraperitoneal injection with 80 mg·kg-1·d-1 STZ high-glucose, high-fat diet. The mice were divided into normal group (normal saline), model group (normal saline) and QWBZS group (18.7 g·kg-1·d-1). After 28 days, the hypoglycemic effect of the drug and its effect on serum total cholesterol (T-CHO), fasting insulin (FINS) and serum tumor necrosis factor-α (TNF-α) were determined. Western blot and Real-time fluorescence quantitative PCR (Real-time PCR) were used to detect protein and mRNA expressions of insulin receptor (IR), insulin receptor substrate-1 (IRS-1)，phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt) in liver tissues．ResultA total of 36 active components in this drug were identified by network pharmacology. KEGG analysis suggested that QWBZS might play a role in reducing blood glucose by regulating PI3K Akt signal pathway. Compared with the model group, the levels of blood glucose, serum T-Cho and TNF-α of the intervention group were significantly lower (P<0.01), while the FINS of the intervention group was significantly higher (P<0.01). Protein and mRNA expressions of IR，IRS-1，PI3K and Akt in liver tissues of mice in QWBZS treatment group increased markedly (P<0.05，P<0.01).ConclusionQWBZS could regulate the levels of blood glucose, TNF-α, T-CHO, and FINS in the serum of diabetic mice induced by high-fat diet and STZ. It can improve PI3K/Akt signal pathway of diabetic mice by regulating protein and mRNA expressions of IR，IRS-1，PI3K and Akt/PKB.
Abstract：ObjectiveTo explore the mechanism of modified Guipitang in the treatment of Yin-Fire insomnia with anxiety with the help of network pharmacological analysis technology.MethodTraditional Chinese Medicine Systems Pharmacology (TCMSP) was used to screen the main components and target genes of modified Guipitang. GeneCards and Online Mendelian Inheritance in Man (OMIM) were used to establish the target gene sets of insomnia and anxiety. STRING 11.0 software was used to analyze the interaction between the overlapping genes, and Cytoscape_3.6.1 software analysis and Matthews correlation coefficient (MCC) algorithm were used to screen the core genes. Based on the results of network analysis, 48 SD female rats were randomly divided into blank control group, model group, eszopiclone tablets group (0.2 mg·kg-1·d-1), modified Guipitang low，medium，and high-dose groups (0.31，1.25，5 g·kg-1·d-1). The model of insomnia with anxiety was established by intraperitoneal injection of Para-chlorophenylalanine (PCPA) and these rats were treated with corresponding drugs for 7 days. Then the frequency, time and distance of the activities were observed in the experiment of autonomic activity. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expressions of proactivated protein kinase 8 (MAPK8), RAC-alpha serine/threonine protein kinase (Akt1), mitogen-activated protein kinase 3 (MAPK3) and interleukin-6 (IL-6) in rat hippocampus.ResultA total of 228 active compounds were screened from TCMSP database and 181 intersecting genes of diseases and drugs were obtained by comparing with GeneCards and OMIM comprehensive database. 9 core genes, including MAPK3, MAPK8, Akt1 and IL-6 were identified by STRING software and MCC algorithm. Animal experiments showed that the number of activity times, time and distance of modified Guipitang in high and medium dose groups were significantly lower than those in the model group. The high and middle dose groups of modified Guipitang could significantly inhibit the mRNA expression of MAPK3, MAPK8, Akt1 and IL-6 in hippocampus（P<0.01）, while the low dose group had no significant effect.ConclusionThe mechanism of modified Guipitang in treating Yin-fire insomnia with anxiety may be related to the regulation of MAPK3, MAPK8, Akt1 and IL-6 genes.
Abstract：ObjectiveTo analyze the research status of gouty arthritis at home and abroad in the past 15 years，and to provide reference for for further in-depth research in this field.MethodWith “Gouty arthritis” as the research topic, the related papers during 2004—2019 were searched in Web of science core collection database and CNKI databases respectively. The results were statistically sorted out according to publication year，author，institution，literature source and key words，etc，and were visualized by CiteSpace software.ResultA total of 5 071 Chinese papers and 1 136 English papers were included. The amount of domestic and foreign publications continued to rise，forming a core team represented by LI Zhao-fu，XIONG Hui，SCHLESINGER NAOMI and other authors. The research hotpots focused on the pathogenesis，diagnosis methods， Chinese and Western medicine treatment，clinical observation，risk assessment and other aspects of gouty arthritis. Both of them had their own emphases. Domestic researches tended to focus on the treatment of gouty arthritis with traditional Chinese medicine，while foreign ones focused on the pathological research and clinical investigation of gouty arthritis.ConclusionThe number of researches in the field of gouty arthritis is on the rise as a whole，and there are both consistency and differences in the research content and hot topics between domestic and foreign literature. Therefore， we should strengthen cooperation and exchange between different teams and countries，so as to promote the overall development of this field.
Abstract：Foeniculi Fructus is warm and spicy in nature and belongs to the liver, kidney, spleen and stomach meridians with the function of dispersing cold and relieving pain, regulating Qi-flowing and harmonizing stomach, and can be used as medicine and food. This article summarized the historical evolution of processing of Foeniculi Fructus by consulting the relevant books since the Han dynasty, the Chinese Pharmacopoeia as well as the processing norms of provinces and cities, and summarized the chemical compositions and pharmacological effects of Foeniculi Fructus by reviewing literatures at home and abroad, then it found that Foeniculi Fructus mainly contains essential oil, flavonoids, phenols and fatty acids, has certain effects on human visceral system, central nervous system, cardiovascular system, endocrine system, immune system and chemotherapy, including the functions of regulating gastrointestinal function, analgesia, anti-inflammatory, etc. Among them, the main chemical components that play the pharmacodynamic effect are volatile oil, flavonoids and phenols, and the chemical components and pharmacological effects in Foeniculi Fructus before and after processing will have corresponding changes, which can provide reference for the further study of Foeniculi Fructus in processing principle, pharmacodynamic substance basis and pharmacological mechanism.
Abstract：Protein phosphorylation is one of the main ways to activate protein bioactivity and make it participate in cell life activities. Researches have shown that approximately 30% of proteins in the human body are modified by phosphorylation at different levels and at different sites. If the protein phosphorylation modification level or site is abnormal, it will cause the occurrence and development of malignant tumors. Malignant tumors have always been a kind of diseases that endanger human life and health. According to statistics, as many as 18 million malignant tumors and more than 9.6 million deaths occur every year worldwide. With the continuous recognition on the abnormality of protein phosphorylation modification in tumorigenesis and development, the research and development of drugs for abnormality of protein phosphorylation modification has become a focus in the field of tumor therapy at present. Each traditional Chinese medicine(TCM) can be regarded as a natural molecular library, and it participates in the regulation of protein phosphorylation modification level with the advantages of multiple components and multiple targets, with slight side effect and low drug resistance, so TCM is one of the main sources of drug development for regulating protein phosphorylation modification levels. Through the search of multiple databases at home and abroad, it was found that certain monomers, parts extracted from TCM, single-TCM and TCM compounds can affect the tumor progression by regulating the level of protein phosphorylation and exert better anti-tumor effect. Based on the current research status of protein phosphorylation regulation by TCM at home and abroad, we found that TCM can inhibit tumor cell proliferation, invasion and metastasis, angiogenesis, and maintenance of stem cell stemness by regulating protein phosphorylation levels, and exert antitumor effects by promoting apoptosis. In order to clarify the molecular mechanism of TCM in regulating protein phosphorylation level and exerting antitumor effect, and provide evidences for the development and clinical application of antitumor TCM, the authors reviewed the mechanism of TCM in regulating protein phosphorylation level and exerting their antitumor effect from different ways in this paper.
Keywords：tumor;protein phosphorylation;traditional Chinese medicine;proliferation;apoptosis;tumor stem cells
Abstract：The pathological mechanism of allergic rhinitis (AR) involves hierarchically acute and chronic reactions among antigens, immunocytes, immunoglobulins and inflammatory factors. AR, which is characterized by heterogeneity, reacts differently to the current treatment on diverse individuals. Some patients receive unsatisfying clinical outcomes after the treatment. Chinese herbal medicine has become a frequent option to treat AR because it can improve clinical symptoms with excellent safety. The description of Biqiu in ancient Chinese medical literatures is very similar to the clinical manifestation of AR. The major pathogeneses of Biqiu included infection of wind-cold pathogens, deficiency and cold in the lung, and evil depression into heat and wind. Therefore, in modern traditional Chinese medicine （TCM）, Biqiu is considered as an integration of AR, allergic sinusitis and vasomotor rhinitis. According to the pathogenesis and clinical manifestations of Biqiu (including AR), this disease belongs to wind disease in TCM. But modern TCM focuses on deficiency of internal organs and ignores wind evil in syndrome differentiation, contrary to the fact that wind medicine is always used in treating AR. This even leads to an abnormal phenomenon that clinical studies attach little importance to syndrome differentiation. In this paper, we started from the phenomenon that wind medicine is always used in TCM for AR, considering TCM literatures and the study results of modern therapy for wind diseases, and put forward the ideas as follows: AR belongs to exogenous wind disease in TCM, and wind pathogen is the major pathogen, in clinical syndrome differentiation, we should attach more importance to wind evil, and dispelling wind and eliminating dampness is the key to treatment. Meanwhile, based on the fact that most of the classical prescriptions with good effectiveness for AR originated from Treatise on Febrile Disease, mainly including the wind medicines for treating exogenous diseases, it can be inferred that the wind-dispelling and itch-arresting effects of wind medicines are helpful to alleviate or block the allergic reaction of nasal mucus and achieve the goal of relieving itching and sneezing, and the wind-dispelling and dampness-eliminating effects are helpful to alleviate the edema of nasal mucus and achieve the goal of relieving running nose and nasal obstruction. In modern researches, it is also found that classical prescriptions which take wind medicine as a principle component and wind medicine used in AR treatment have good anti-inflammatory, anti-allergic and immune function adjusting effects. This indicates that dispelling wind and eliminating dampness and the wind medicine treatment under its instruction not only correspond to TCM theory, but also have modern science evidences.
Keywords：dispelling wind and eliminating dampness therapy;allergic rhinitis;wind disease;wind-dispelling medicine
Abstract：Tumor has become the second most serious disease that threatens human health and life. Treating with chemical drugs (referred to as chemotherapy) is the most basic treatment, but most chemotherapeutic drugs cause damage to normal tissues. It is a difficult problem in the field of biomedical research that how to deliver anti-tumor drugs more efficiently, increase the concentration of drugs in tumor tissues, enhance the anti-tumor effect, and decrease the drug distribution in normal tissues to weaken the damage to normal tissues. In order to achieve the goals of accurate delivery of anti-tumor drugs and synergism and attenuation, the researchers used systematic evolution of ligands by exponential enrichment technology (SELEX technology) to screen aptamers that can specifically target tumor markers or tumor cells, and designed the novel liposome targeting drug delivery system with aptamers as targeting molecules (ligands). This paper briefly introduced nucleic acid aptamer technology and common tumor markers, and reviewed the research advances on the antitumor effect of aptamer-liposome drug delivery system. It will provide references for the selection of appropriate tumor markers as targets and the application of aptamer technology in the research and development of high-efficiency and low-toxicity liposome targeting agents of anti-tumor traditional Chinese medicine. Meanwhile, it is of great significance for promoting the application of aptamer technology in targeted drug delivery systems.
Keywords：aptamer-liposome targeted drug delivery system;tumor markers;nucleic acid aptamer technology;liposomes;anti-tumor;systematic evolution of ligands by exponential enrichment technology
Abstract：Dyslipidemia is a disease of lipid metabolism. At present, the prevalence of dyslipidemia in adults in China is as high as 40.40%. In the United States, there are more than 100 million individuals with abnormal low-density lipoprotein cholesterol (LDL-C), and the incidence rate is increasing year by year and showing a trend of becoming younger. Dyslipidemia is closely related to a variety of diseases such as fatty liver, atherosclerosis , hypertension, coronary heart disease, diabetes, and stroke. It has now developed into a global public health problem that seriously threatens human life and health. Modern medicine believes that its pathogenesis is complicated and is related to abnormal glucose and lipid metabolism, insulin resistance (IR) and other factors. Chinese medicine ascribes it to primary asthenia-secondary sthenia syndrome, which is closely related to the liver, spleen, and kidney. It is believed that excessive fat and grease can cause phlegm and cause many diseases. In terms of its treatment, western medicine mainly uses statin chemical synthesis preparations, with stable therapeutic effect, but many adverse reactions such as myalgia, myositis, rhabdomyolysis and acute renal injury are the main factors restricting its clinical application. Traditional Chinese medicine (TCM) has a long history, and multi-pathway, multi-target, multi-level regulation of dyslipidemia, few adverse reactions and low drug dependence are the principal advantages of TCM in treating dyslipidemia. At present, there are more and more researches on the prevention and treatment of dyslipidemia by TCM, but they are mainly focused on the observation of curative effect and the summary of prescription, and there are relatively few in-depth discussion and summary of the mechanism of TCM. Through comprehensively retrieving and collating the relevant domestic and foreign literatures in the past five years, we reviews from the perspective of effective ingredients, therapeutic pathways, and targets of action, and comprehensively introduces the latest research progress of TCM on the mechanism of regulating dyslipidemia, and put forward some suggestions for the possible research direction in the future, in order to provide new ideas and theoretical basis for TCM in clinical prevention and treatment of this disease.
Keywords：dyslipidemia;active ingredients of traditional Chinese medicine;targets;mechanism of action;advance
Abstract：Cerebral infarction is a clinical disease with corresponding neurological symptoms caused by cerebral ischemia and hypoxia caused by cerebral blood supply disorder. It is one of the most common cerebrovascular diseases and a serious threat to human health. The prevention and treatment of cerebral infarction has an important social significance. Angiogenesis is the key starting point for medical treatment of cerebral infarction, and signal transduction and transcriptional activators (STAT)/hypoxia inducing factor-1(HIF-1)/vascular endothelial growth factor (VEGF) pathway are important pathways to mediate angiogenesis after cerebral infarction. This paper took the angiogenesis as the starting point and the upstream molecules of STAT/HIF-1/VEGF signal pathway STAT3 and miRNA as the main study objects, and comprehensively discussed the results of chip sequencing, experimental research, traditional Chinese medicine (TCM) pathogenesis and TCM treatment. Based on the regulatory mode of "TF-miRNA" and the idea of "micro-whole", it is suggested that under the guidance of the basic theory of TCM, cubic compound prescriptions of TCM and its active components might activate the STAT/HIF-1/VEGF signal pathway through STAT3/miRNA feedback loop to promote angiogenesis after cerebral infarction, which puts forward a deep molecular mechanism and new direction for the treatment of cerebral infarction with TCM.
Keywords：cerebral infarction;angiogenesis;signal transduction and transcriptional activators3 (STAT3);miRNA;traditional Chinese medicine
Abstract：Diarrhea-predominant irritable bowel syndrome (IBS-D) is one of the common functional gastrointestinal diseases in clinical practice, its pathogenesis is diverse. Because of its sudden and lingering intractable symptoms, it seriously affects patients' work and life. IBS-D patients suffer from repeated illnesses, which often affect their lives with mental symptoms such as anxiety and depression. Anxiety and depression can also affect visceral sensation, increase intestinal sensitivity, aggravated by interaction between physical symptoms and mental symptoms. The main pathogenesis of IBS-D such as visceral hypersensitivity, gastrointestinal motility disorders, intestinal infections, and psychosocial factors are all related to brain-gut interaction disorders. Patients with IBS-D are prone to brain-gut interaction disorders due to long-term chronic mental stress. Brain-gut interaction is the main mode of regulation of gastrointestinal function in the brain-gut axis. Traditional Chinese medicine （TCM） believes that the main syndrome type of IBS-D patients is liver stagnation and spleen deficiency, while liver stagnation will be unsatisfactory, and the secretion and content of various neurotransmitters in the brain are closely related to emotions. Tongxie Yaofang is a commonly basic prescription used for the clinical treatment of IBS-D liver stagnation and spleen deficiency syndrome. It has the functions of softening the liver and replenishing the spleen, removing dampness and stopping diarrhea, and has a significant clinical effect. At present, many animal experiments and clinical studies have explored the mechanism of Tongxie Yaofang in treating IBS-D and its effect on brain-gut interaction function and brain-gut peptide content, but the main liver drainage and brain-gut interaction have not been linked. The author took the correlation between liver liver govers regulating and brain-gut interaction as the starting point, explored the mechanism of Tongxie Yaofang in treating IBS-D based on brain-gut interaction disorder, and explained the correlation between the three. Based on the research literature of Tongxie Yaofang in the past 5 years and the interaction between Tongxie Yaofang and brain-gut interaction, the author explored the effect of Tongxie Yaofang in treating IBS-D by affecting the brain-gut axis and brain-gut peptides.