Abstract：ObjectiveTo explore the mechanism of Sishenwan， Baitouweng Tang， and Lianlitang in the treatment of ulcerative colitis （UC）， and compare their efficacies on UC in rats.MethodNinety SD rats of SPF grade were randomly divided into blank group （distilled water， 2 mL·d-1） and experimental group. The rats in the experimental groups were administered with 2，4，6-trinitrobenzene sulfonic acid （TNBS） by clysis to induce the UC model. Subsequently， the model rats were divided into a model group （distilled water， 2 mL·d-1）， positive group ［sulfasalazine （SASP）， 0.4 g·kg-1·d-1］， Sishenwan group （1.76 g·kg-1·d-1）， a Baitouweng Tang group （1.40 g·kg-1·d-1）， and Lianlitang group （2.13 g·kg-1·d-1） according to the random number table. The rats in each group were dosed at 2 mL·d-1 for 14 days. The pathological score for colonic mucosa was detected. Cytokines were detected by the cytokine chip. The enzyme-linked immunosorbent assay （ELISA） was used to detect the free triiodothyronine （FT3）， glucagon-like peptide-1 （GLP-1）， and corticosterone （CORT） in plasma， and neurotensin （NT）， substance P （SP）， vasoactive intestinal peptide （VIP）， and somatostatin （SST） in colon tissues.ResultCompared with the normal group， the model group showed increased colon mass-length ratio and pathological score for colonic mucosa （P<0.01）， infiltration of massive lymphocytes， disordered or absent intestinal villi， elevated levels of cytokine-induced neutrophil chemoattractant-1/2α/β/3 （CINC-1/2α/β/3）， interleukin-1α （IL-1α）， interferon-inducible protein-10 （IP-10） and other factors in colon tissues （P<0.05）， dwindled CORT and GLP-1 levels in plasma （P<0.05）， and increased SP content in colon tissues （P<0.05）. Compared with the results in the model group， the mucosal injury in the colon of rats in each drug group was relieved. The levels of IL-1α， IP-10， lipopolysaccharide-inducible CXC chemokine （LIX）， and L-selectin of rats in the Lianlitang group and Sishenwan group were reduced （P<0.05）， and the CINC-3 and IL-17 levels were diminished in the Baitouweng Tang group （P<0.05）. The levels of CINC-1/3， IL-1α， and IP-10 were reduced in the SASP group （P<0.05）. The plasma FT3 was up-regulated in the Lianlitang group， and the plasma GLP-1 levels were elevated in the three Chinese medicine groups （P<0.05）. The VIP content in colon tissues of the Sishenwan group and Baitouweng Tang group was down-regulated （P<0.05）， and the SST content in colon tissues of the SASP group was significantly up-regulated （P<0.01）.ConclusionThe intervention of Lianlitang and Sishenwan on UC was significant， and the underlying mechanism of action might be related to inflammation inhibition and immune balance by regulating the cytokine network. The efficacy of Lianlitang was predominant， followed by Sishenwan and Baitouweng Tang.
Abstract：ObjectiveTo explore the effect of hypoxia-inducible factor （HIF）-1α on T helper 17 （Th17）/regulatory T cell （Treg） balance in ulcerative colitis and the intervention mechanism of Shaoyaotang.MethodForty-eight SD rats were randomly divided into normal group （normal saline）， model group， mesalazine group （0.42 g·kg-1）， Shaoyaotang group （11.1 g·kg-1）， inhibitor group ［2-methoxyestradiol （2ME2）， 0.015 g·kg-1］， and Shaoyaotang+inhibitor group. The ulcerative colitis model was induced by 2，4，6-trinitrobenzene sulfonic acid （TNBS）. The rats in all groups received corresponding treatments for 7 d， and the general condition and disease activity index （DAI） were observed. Hematoxylin-eosin （HE） staining was used to observe histopathological changes of the colon. Enzyme-linked immunosorbent assay （ELISA） was employed to detect serum levels of interleukin （IL）-10， IL-17， and IL-23 in rats. Western blot was used to detect the expression levels of forkhead box protein 3 （FoxP3）， retinoic acid-related orphan receptor γt （RORγt）， and HIF-1α proteins in the colon tissue.ResultCompared with the normal group， the model group showed elevated disease activity index （DAI） score and pathological score for intestinal mucosa （P<0.01）， reduced serum IL-10 level （P<0.01）， up-regulated IL-17 and IL-23 levels （P<0.01）， increased RORγt and HIF-1α expression （P<0.01）， and decreased FoxP3 protein expression （P<0.01）. Compared with the model group， the Shaoyaotang group displayed diminished DAI score and pathological score for intestinal mucosa （P<0.05， P<0.01）， increased serum IL-10 level （P<0.01）， decreased IL-17 and IL-23 levels （P<0.01）， dwindled protein levels of RORγt and HIF-1α （P<0.01）， and up-regulated expression of FoxP3 （P<0.01）. Compared with the inhibitor group， the Shaoyaotang group and the Shaoyaotang+inhibitor group exhibited significant differences in the expression of RORγt， FoxP3， and HIF-1α proteins （P<0.05， P<0.01）.ConclusionShaoyaotang could effectively treat ulcerative colitis， and the underlying mechanism of action might be related to the regulation of Th17/Treg rebalance by inhibiting HIF-1α.
Keywords：ulcerative colitis（UC）;Shaoyaotang;T helper 17 （Th17）;regulatory T cell （Treg）;hypoxia-inducible factor-1α （HIF-1α）
Abstract：ObjectiveConsidering the efficacy of Gegen Qinliantang （GQT） in releasing exterior and clearing interior to alleviate dampness-heat dysentery， we analyzed the mechanism of the chloroform extract of GQT in alleviating enterotoxicity caused by irinotecan to provide an experimental basis for the development of GQT.MethodKunming mice （n=60） were randomly divided into a blank group， a model group， a loperamide group （positive drug of loperamide hydrochloride capsule， 0.4 mg·kg-1）， and high- （2.3 g·kg-1） and low-dose （1.16 g·kg-1） GQT chloroform extract groups. The mouse model of delayed diarrhea was established by intraperitoneal injection of irinotecan hydrochloride （CPT-11， 55 mg·kg-1） for four consecutive days， meanwhile， the mice in the blank group only received the same volume of normal saline. Corresponding drugs were administered by gavage on the fifth day， respectively， while the ones in the blank group and model group were given distilled water for five consecutive days. The general condition of mice in each group was observed， and diarrhea indexes of mice were recorded. Pathological changes in colon tissues of mice were observed by hematoxylin-eosin （HE） staining. The tumor necrosis factor （TNF）-α， interleukin （IL）-1β， cyclooxygenase （COX）-2， intercellular adhesion molecule （ICAM）-1， glutathione peroxidase （GSH-Px）， superoxide dismutase （SOD）， malondialdehyde （MDA） and nitric oxide （NO） levels in colon tissues were detected with the assay kits. Furthermore， the expression levels of Kelch sample epoxy chloropropane associated protein 1 （Keap1）， nuclear factor E2 related factor 2 （Nrf2）， tight junction protein-1 （ZO-1）， heme oxygenase-1 （HO-1） and tight junction protein （Occludin） were detected by Western blot.ResultCompared with the blank group， the model group showed declined body weight and reduced contents of GSH-Px and SOD （P<0.01）， whereas increased diarrhea indexes and TNF-α， IL-1β， COX-2， ICAM-1， MDA and NO levels （P<0.01）. Abundant inflammatory cells and colonic mucosa with defects， swelling， bleeding， and inflammatory exudation were revealed by HE staining in the mice of the model group. The expression levels of Keap1， Nrf2， ZO-1， HO-1 and Occludin in colon tissues significantly declined （P<0.01）. Compared with the model group， the loperamide group and the high- and low-dose GQT chloroform extract groups exhibited improved weight loss， reduced diarrhea indexes， diminished TNF-α， IL-1β， COX-2， ICAM-1， MDA and NO， and elevated GSH-Px and SOD. HE staining indicated that the cells were compactly arranged with clear nuclei in the high- and low-dose GQT chloroform extract groups， and the expression levels of Keap1， Nrf2， HO-1， Occludin， and ZO-1 were up-regulated.ConclusionGQT chloroform extract may alleviate CPT-11-induced delayed diarrhea by regulating inflammation and oxidative stress for enhancing the intestinal barrier function. These findings are expected to provide a reference for exploring the toxicity-attenuating effect of Chinese medicinals on chemotherapy drugs and for developing famous classical formulas.
Abstract：ObjectiveTo explore the quality transmitting relationship between decoction pieces and substance benchmarks with the fingerprint， index component content and dry extract rate as evaluation indexes， and investigate the key quality attributes of 15 batches of substance benchmarks of Yihuangtang， and establish the quality standard of this substance benchmarks.MethodFifteen batches of Yihuangtang substance benchmarks freeze-dried powder samples were prepared， the fingerprint and index component content of 15 batches of decoction pieces and substance benchmarks were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A）-0.1% phosphoric acid aqueous solution （B） for gradient elution （0-6 min， 97%B； 6-12 min， 97%-92%B； 12-25 min， 92%-90%B； 25-35 min， 90%-89%B； 35-50 min， 89%-82%B； 50-75 min， 82%-72%B； 75-85 min， 72%-35%B）， the detection wavelength was set at 230 nm， combined the dry extract rate to clarify the attribution of characteristic peaks and the range of similarity with the control chromatogram， the content range and transfer rate range of geniposidic acid and berberine hydrochloride， the dry extract rate range and the variation range of the substance benchmarks.ResultThe established HPLC fingerprint had good precision， repeatability and stability， and could be used for the simultaneous determination of decoction pieces and substance benchmarks of Yihuangtang. The similarities between the control chromatogram and fingerprint of substance benchmarks were >0.99. A total of 15 characteristic peaks were assigned， and 8 characteristic peaks were identified by the reference substances， of which 6 were from Phellodendri Chinensis Cortex processed with salt， 1 was from Plantaginis Semen processed with wine， and 1 was from stir-fried Dioscoreae Rhizoma. The content ranges of geniposidic acid and berberine hydrochloride in 15 batches of substance benchmarks of Yihuangtang were 0.10%-0.16% and 0.63%-1.05%， the transfer rate ranges of them were 20.91%-32.65% and 19.60%-29.59%， respectively. The dry extract rate range of the substance benchmarks was 8.45%-9.92%.ConclusionThe quality standard of Yihuangtang substance benchmarks can be preliminarily formulated by the combination of fingerprint， dry extract rate and determination of index component， which can provide the basis for the quality control of Yihuangtang and the development of related preparations.
Abstract：Jichuanjian，a famous classical formula with definite clinical efficacy in Jingyue Quanshu （《景岳全书》） by ZHANG Jing-yue in the Ming Dynasty， was included in the Catalogue of Ancient Famous Classical Formulas （The First Batch） published by the National Administration of Traditional Chinese Medicine（TCM） in 2018．By means of bibliometrics，43 relevant ancient book data were collected，including 26 ancient books of TCM. The historical origin and development，main indications and symptoms，composition and prescription，dosage，processing，and decoction and administration methods of Jichuanjian were systematically summarized to provide literature support for the development and clinical application of famous classic formulas. It was found that the main symptoms of Jichuanjian were expanded in the past dynasties from deficiency constipation， postpartum constipation， and qi stagnation recorded in the original formula to additional dryness accumulation， blood stasis， Yin constipation， infantile constipation， and lung（intestine）diseases induced by eye diseases. The applicable population of Jichuanjian was extended to children in addition to puerpera and adults，and the disease scope was extended to pediatrics， ophthalmology， and otorhinolaryngology in addition to internal medicine，gynecology and obstetrics．Although Jichuanjian was modified by doctors in the past dynasties，the compatibility of Angelicae Sinensis Radix，Achyranthis Bidentatae Radix，Cistanches Herba，Alismatis Rhizoma，Cimicifugae Rhizoma and Aurantii Fructus has been constant. The decoction and administration methods basically remained unchanged， i.e.， water decoction and administration before meals. Meanwhile， doctors in the Qing Dynasty made a lot of general comments on this formula with little controversy．
Keywords：famous classical formula;Jichuanjian;change in formula composition;textual research
<CITATION><Bulk Citation>11552437 false
Study on Treatment of Senile Chronic Sleep Deprivation Rats with Suanzaoren Tang
Abstract：ObjectiveTo study the mechanism of Suanzaoren Tang in regulating the energy metabolism of myocardial mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation through the sirtuin 3 （SIRT3）/superoxide dismutase2 （SOD2） signaling pathway.MethodFifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg-1·d-1）， and low- （6.48 g·kg-1·d-1） and high-dose （12.96 g·kg-1·d-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of D-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of myocardial mitochondria was observed under a transmission electron microscope. The content of adenosine triphosphate （ATP） in rat hypothalamus was detected by colorimetry， while the malondialdehyde （MDA） content and the SOD activity in myocardium were measured by spectrophotometry. Real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to determine the mRNA and protein expression levels of SIRT3 and SOD2 in rat myocardium. The localization of SIRT3 was detected by immunofluorescence staining.ResultCompared with the control group， the model group exhibited a disordered arrangement of myocardial filaments， accompanied by filament rupture and dissolution， obviously swollen mitochondria arranged in disorder， and blurring and even rupture of most mitochondrial cristae. Besides， the content of ATP and SOD activity in the myocardium decreased significantly （P<0.01）， whereas that of MDA increased significantly （P<0.01）. The mRNA and protein expression levels of SIRT3 and SOD2 were down-regulated significantly （P<0.01）， and the average fluorescence intensity of SIRT3 protein declined significantly （P<0.01）. The comparison with the model group revealed that high-dose Suanzaoren Tang enabled the myocardial filaments to be neatly arranged， relieved the mitochondrial damage and swelling， only manifested as partial mitochondrial cristae rupture， significantly increased ATP content， SOD activity， as well as SIRT3 and SOD2 mRNA and protein expression levels （P<0.01）， reduced the content of MDA （P<0.01）， and enhanced the average fluorescence intensity of SIRT3 protein （P<0.05）. The myocardial mitochondrial injury in the estazolam group was also alleviated. The activity of SOD and the SIRT3 and SOD2 mRNA and protein expression levels in the myocardium were significantly elevated （P<0.01）， while the activity of MDA was significantly lowered （P<0.01）. In the low-dose Suanzaoren Tang group， the improvement in myocardial mitochondrial injury was not obvious. However， both the SOD activity and SOD2 protein expression were significantly increased （P<0.05）.ConclusionSuanzaoren Tang ameliorates the myocardial mitochondria injury and abnormal energy metabolism induced by chronic REM sleep deprivation in aged rats possibly by up-regulating the SIRT3 and SOD2 expression.
Keywords：Suanzaoren tang;sleep deprivation;transmission electron microscope;energy metabolism
Abstract：ObjectiveTo investigate the effect of Suanzaoren Tang on mitochondria-mediated neuronal apoptosis.MethodFifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg-1·d-1）， and low- （6.48 g·kg-1·d-1） and high-dose （12.96 g·kg-1·d-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of D-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of mitochondria in hypothalamus was observed under a transmission electron microscope. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in hypothalamus were detected by spectrophotometry. Western blotting was conducted to determine the protein expression levels of cytochrome C （Cyt C）， B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax） and cysteinyl aspartate-specific protease-3 （Caspase-3） in hypothalamus.ResultIn the control group， there was no obvious pathological change in mitochondria， which were moderate in size and oval or spindle in shape， with the cristae arranged orderly. Compared with the control group， the model group exhibited abnormal mitochondrial morphology， manifested as obvious swelling， vacuolation， myelin figures， and cristae rupture and reduction. The comparison with the model group revealed that both the estazolam group and high-dose Suanzaoren Tang group alleviated the mitochondrial damage and reduced the vacuolation and swelling. Only some cristae rupture was present. The improvements were more obvious in the high-dose Suanzaoren Tang group. Compared with the control group， the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase and the Bcl-2 protein expression in the model group were significantly decreased （P<0.01）， whereas the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly increased （P<0.01）. Compared with the model group， the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase and the Bcl-2 protein expression in the estazolam group and the high-dose Suanzaoren Tang group were significantly elevated （P<0.01）， while the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly lowered （P<0.05， P<0.01）. The activity of Na+-K+-ATPase and the Bcl-2 protein expression in the low-dose Suanzaoren Tang group were increased significantly （P<0.01）， but the Bax protein expression was down-regulated （P<0.05）.ConclusionSuanzaoren Tang is able to improve the mitochondrial function of hypothalamic nerve cells and inhibit their apoptosis.
Keywords：Suanzaoren tang;sleep deprivation;transmission electron microscope;energy metabolism
Abstract：ObjectiveTo study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation.MethodFifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg-1·d-1）， and low- （6.48 g·kg-1·d-1） and high-dose （12.96 g·kg-1·d-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of D-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of liver mitochondria was observed under the transmission electron microscope. The content of adenosine triphosphate （ATP） in rat liver was detected by colorimetry， and the activities of ubiquinone oxidoreductase （complex Ⅰ）， succinate-ubiquinone oxidoreductase （complex Ⅱ）， ubiquinol-cytochrome c oxidoreductase （complex Ⅲ）， and cytochrome c oxidase （complex Ⅳ） in mitochondrial respiratory chain of rat liver were measured by colorimetry. The protein expression levels of citrate synthase （CS）， isocitrate dehydrogenase （IDH）， and ATP synthase， H+ transporting， mitochondrial F0 complex， subunit b， isoform 1 （ATP5F1） in rat liver were assayed by Western blot.ResultThe mitochondrial damage in rat liver of the model group was more serious than that in the control group， manifested as mitochondrial swelling and deformation as well as cristae rupture and reduction. The comparison with the model group revealed that both the positive control and Suanzaoren Tang at the high dose obviously alleviated the mitochondrial swelling and deformation and reduced cristae rupture， with better improvements observed in the high-dose Suanzaoren Tang group. Compared with the control group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the model group were all significantly decreased （P<0.01）. Compared with the model group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the high-dose Suanzaoren Tang group were all significantly increased （P<0.05，P<0.01）. In the positive control group， the content of ATP， the activities of mitochondrial respiratory chain complexes I and Ⅲ， and the protein expression levels of CS and ATP5F1 in rat liver were significantly increased （P<0.05， P<0.01）. The activities of mitochondrial respiratory chain complexes Ⅰ and Ⅲ and the ATP5F1 protein expression in the low-dose Suanzaoren Tang group were significantly elevated （P<0.05， P<0.01）.ConclusionSuanzaoren Tang alleviates the abnormal liver energy metabolism induced by chronic REM sleep deprivation in the elderly rats， which may be related to its enhancement of mitochondrial electron transport chain enzyme activities and the up-regulation of protein expression levels of CS， IDH and ATP5F1.
Keywords：Suanzaoren tang;sleep deprivation;transmission electron microscope;liver;energy metabolism
Abstract：Sleep has been widely concerned by the medical field all over the world. Sleep deprivation can cause damage to organs of the human body， which is related to the occurrence of a variety of diseases. Besides， the pathological change in different organs of the human body is also a key factor that causes or aggravates insomnia. When treating insomnia and its complications， traditional Chinese medicine （TCM） focuses on the homology of the brain and heart， and insomnia is mainly treated from the five internal organs， especially the heart and liver. Sleep duration and structure change with age. The prevalence of insomnia is higher in older individuals susceptible to complications than in the younger population. In TCM， insomnia of blood deficiency and Yin deficiency is common among the elderly. Suanzaoren Tang is a classic prescription for nourishing blood and calming the mind and it is critical in the treatment of "sleeplessness due to consumptive disease and dysphoria"， with the effects of nourishing liver blood to calm the mind and clearing internal heat to relieve dysphoria. It has good efficacy on the insomnia of the elderly caused by deficiency of Qi and blood and abnormal operation of nutrient Qi and defense Qi. Furthermore， it also shows a certain therapeutic effect on insomnia combined with cardiovascular and cerebrovascular diseases. The present study revealed the damage to the brain， heart， and liver caused by sleep deprivation and the effect of Suanzaoren Tang on the brain， heart， and liver， and clarified the facts that Suanzaoren Tang inhibited the damage to organs caused by sleep deprivation and regulated energy metabolism， thereby exploring the sedative and hypnotic mechanism of Suanzaoren Tang to provide new ideas for Suanzaoren Tang in the treatment of sleep disorders and other diseases.
Abstract：ObjectiveTo investigate the protective effect and molecular mechanism of Angelicae Sinensis Radix-Chuanxiong Rhizoma medicated serum （ASRCRS） against oxidative damage of PC12 cells induced by H2O2.MethodOxidative damage of PC12 cells was induced by H2O2 in vitro， and intervention was performed in the low-， medium-， and high-dose ASRCRS groups with a final volume fraction of 15%. The cell viability was determined by methyl thiazolyl tetrazolium （MTT） assay. Cell morphology was observed by an inverted fluorescence microscope. The content of lactate dehydrogenase （LDH） and malondialdehyde （MDA）， the activity of superoxide dismutase （SOD）， and the distribution of reactive oxygen species （ROS） in the cell supernatant were detected by the kits. Cell apoptosis was detected by Annexin V-FITC/PI double staining. The protein expression levels of nuclear factor E2-related factor 2 （Nrf2）， Kelch-like epichlorohydrin associated protein-1 （Keap1）， heme oxygenase-1 （HO-1）， and SOD1 were detected by Western blot.ResultOxidative damage was induced by 300 μmol·L-1 H2O2 for 24 hours. Compared with the normal group， the model group showed abnormal cell morphology， reduced cell viability （P<0.01）， increased LDH and MDA （P<0.01）， blunted SOD activity， elevated intracellular distribution of ROS， down-regulated protein expression of Nrf2， HO-1， and SOD1 （P<0.05， P<0.05）， and up-regulated protein expression of Keap1 （P<0.01）. Compared with the model group， ASRCRS groups displayed improved cell morphology， increased cell viability， inhibited cell apoptosis， potentiated SOD activity （P<0.01）， suppressed release of LDH （P<0.01） and generation of ROS， decreased content of MDA （P<0.01）， up-regulated protein expression of Nrf2， HO-1 and SOD1 （P<0.05， P<0.01）， and down-regulated protein expression of Keap1 （P<0.01）.ConclusionASRCRS could protect PC12 cells from oxidative damage induced by H2O2 by up-regulating the expression of Nrf2 to activate the Nrf2/antioxidant response element （ARE） signaling pathway， enhancing the ability to resist oxidative damage， and inhibiting cell apoptosis.
Abstract：ObjectiveTo investigate the inhibitory effect of Fuzheng Jiedu prescription（FZJDP） combined with 5-fluorouracil （5-Fu） against postoperative recurrence and metastasis in gastric cancer-bearing mice and explore the possible mechanism based on changes in CD4+ T cells， CD8+ T cells， and regulatory T （Treg） cells of tumor microenvironment， endoplasmic reticulum stress （ERS） and phosphatidylinositol-3-kinase （PI3K）/protein kinase B（Akt）/mammalian target of rapamycin（mTOR） pathway.MethodForty 615 mice were randomly divided into the model group，FZJDP （25 g·kg-1） group，5-Fu（25 mg·kg-1）group，and combined （25 g·kg-1 FZJDP + 25 mg·kg-1 5-Fu）group， with 10 mice in each group. Mouse forestomach carcinoma （MFC） cells were implanted into the inner side of footpad of the left hind paw and the transplanted tumor was then surgically excised to establish a postoperative recurrence model. Hematoxylin-eosin（HE） staining was conducted to observe the pathological changes in mice with gastric cancer recurrence and lung metastasis. The CD4+/CD8+ T cell ratio in recurrent tumor and the percentages of Treg cells ［CD4+，CD25+， and forkhead box protein P3 （FOXP3）+ cells］ in spleen were detected by flow cytometry. The contents of ERS-related proteins ［78-kDa glucose-regulated protein（GRP78），inositol-requiring enzyme 1 alpha （IRE1α），activating transcription factor 6（ATF6），and protein kinase R-like endoplasmic reticulum kinase（PERK）］ and the expression of related proteins in the PI3K/Akt/mTOR signaling pathway were determined by Western blot and immunohistochemistry （IHC）.ResultCompared with the model group， the combined group significantly increased the recurrent inhibition rate （P<0.05）. The recurrent tumor weight was significantly decreased in each treatment group （P<0.05）. The number of lung metastases and metastasis rate declined in each treatment group， and the lowest values were observed in the combined group， without any statistical significance. The CD4+/CD8+ T cell ratios in the FZJDP group and combined group were significantly elevated （P<0.05）， while the percentages of Treg cells were reduced （P<0.05）. However， 5-Fu resulted in a significant increase in Treg cell percentage （P<0.05）. IHC results showed that the protein expression levels of ATF6 （P<0.05，P<0.01）， IRE1α（P<0.05），and Akt （P<0.01） in each treatment group were significantly down-regulated as compared with those in the model group. As revealed by Western blot， the GRP78 expression level in the 5-Fu group was lower than that in the model group （P<0.05）， and the expression levels of PI3K， phosphorylated Akt （p-Akt）， and mTOR were significantly decreased in the 5-Fu group and the combined group （P<0.05）.ConclusionFZJDP combined with 5-Fu reduces postoperative recurrence and metastasis in tumor-bearing mice possibly by inhibiting PI3K/Akt/mTOR signaling pathway， diminishing ERS，and improving tumor immune microenvironment.
Abstract：ObjectiveTo investigate the regulatory mechanism of Bushen Zhuyun prescription（BSZYP） on endoplasmic reticulum stress （ERS） in rats with luteal phase defect （LPD） induced by mifepristone.MethodFifty SD rats were randomly divided into a blank group， a model group， a positive control group （dydrogesterone，0.02 g·kg-1）， and low-（0.08 g·kg-1）and high-dose （0.24 g·kg-1） BSZYP groups. Western blot and Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） were used to detect the mRNA and protein expression levels of immunoglobulin binding protein （BIP）， protein kinase R-like endoplasmic reticulum kinase （PERK）， inositol-requiring enzyme 1 （IRE-1）， activating transcription factor 6 （ATF6）， and C/EBP homologous protein （CHOP）. The enzyme-linked immunosorbent assay （ELISA） was used to detect the serum progesterone （P） and estradiol （E2） levels.ResultCompared with the blank group， the model group showed the elevated protein expression of BIP， PERK， and CHOP （P<0.01） and the dwindled mRNA expression of PERK and CHOP （P<0.05）， while no significant difference was observed in the protein expression of IRE-1 and ATF6， mRNA expression of IRE-1， BIP， and ATF6， and serum E2 and P levels. Compared with the model group， the positive control group displayed diminished protein expression of CHOP （P<0.01）， while no significant difference was observed in the protein expression of PERK， IRE-1， BIP， and ATF6， mRNA expression of PERK， IRE-1， BIP， ATF6， and CHOP， and serum levels of E2 and P. The protein expression of CHOP decreased （P<0.01） and the mRNA expression of CHOP increased （P<0.05） in the low-dose BSZYP group， while no significant difference was observed in the mRNA and protein expression of PERK， IRE-1， BIP， and ATF6， and serum E2 and P levels. In the high-dose BSZYP group， the protein expression of PERK， BIP， and CHOP was down-regulated （P<0.01）， and the mRNA expression of CHOP was up-regulated （P<0.01）， while no significant difference was observed in the protein expression of IRE-1 and ATF6， mRNA expression of PERK， IRE-1， BIP， and ATF6， and serum E2 and P levels.ConclusionBSZYP can treat LPD by relieving ERS.
Abstract：ObjectiveTo explore the application value of modified Buzhong Yiqitang （BZYQT） in the treatment of postoperative patients with non-small cell lung cancer （Qi deficiency in lung and spleen） after chemotherapy， and to observe its effect on tumor angiogenesis， immune function， tumor indicators， and lung function indicators.MethodNinety-six patients who were treated in the Kunming municipal hospital of traditional Chinese medicine from March 2018 to February 2020 due to postoperative chemotherapy for non-small cell lung cancer were selected and assigned into a control group （n=48， western medicine） and an observation group （n=48， western medicine+modified BZYQT） by the random number table. The curative efficacies were compared after the treatment.ResultAfter treatment， the serum levels of carcinoembryonic antigen （CEA）， cytokeratin 19 fragment 21-1 （CYFRA21-1）， serum insulin-like growth factor-1 （IGF-1）， vascular endothelial growth factor （VEGF）， and transforming growth factor（TGF）-β1 in the observation group were lower than those in the control group （P<0.05）， while the serum CD4+/CD8+，CD4+ cells， immunoglobulin G （IgG） levels， forced expiratory volume in one second （FEV1），and FEV1/forced vital capacity （FVC） in the observation group were higher than those in the control group （P<0.05）. A significant difference was observed in the total response rate between the observation group ［56.25% （27/48）］ and the control group ［35.42% （17/48）］ （χ2＝4.191，P<0.05）. For adverse reactions，the incidence of bone marrow suppression（χ2＝4.002）， gastrointestinal reaction （χ2＝7.069），and hepatic and renal injury （χ2＝5.151） was lower in the observation group than in the control group （P<0.05）.ConclusionFor postoperative patients with non-small cell lung cancer （Qi deficiency in lung and spleen） after chemotherapy， western medicine combined with modified BZYQT could ameliorate immune function， promote pulmonary function recovery， improve clinical efficacy， and reduce the incidence of adverse reactions.
Abstract：ObjectiveTo observe the clinical efficacy of modified Huanglian Wendantang and Shaofu Zhuyutang in the treatment of ovulation disorder in patients with polycystic ovary syndrome （PCOS） due to the combined phlegm and stasis-induced and its influence on chronic inflammation.MethodAccording to the random number table， 100 patients were divided into a control group （50 cases） and an observation group （50 cases）. Apart from lifestyle intervention and oral administration of clomiphene citrate （CC） capsules to induce ovulation， patients in the control group further received Guizhi Fulingwan， 6 g/time， 2 times/day， while those in the observation group were treated with the modified Huanglian Wendantang and Shaofu Zhuyutang， 1 dose/day， for six menstrual cycles. The ovulation， endometrial thickness， proportion of type A endometrium， as well as the pulsatility index （PI） and resistance index （RI） of uterine artery were monitored before and after treatment. The fasting blood glucose （FBG）， fasting insulin （FINS）， luteinizing hormone （LH）， follicle stimulating hormone （FSH）， serum testosterone （T）， estradiol （E2）， dehydroepiandrosterone sulfate （DHEAS）， tumor necrosis factor-α （TNF-α）， high-sensitivity C-reactive protein （hs-CRP）， adiponectin （APN）， and interleukin-6 （IL-6） levels before and after treatment were detected， followed by the calculation of homeostasis model assessment-insulin resistance （HOMA-IR） value and the evaluation of ovarian volume and combined phlegm and stasis-induced syndrome score.ResultThe overall response rate of the observation group was （44/47） 93.62%， which was higher than （36/46） 78.26% of the control group （χ2=4.802， P<0.05）. The ovulation rate in the observation group was （199/264） 75.38%， higher than （173/272） 63.60% in the control group （χ2=8.714， P<0.01）. The clinical pregnancy rate of the observation group was （11/47） 23.40%， higher than （5/46） 10.87% of the control group， but the difference was not statistically significant （χ2=2.564， P>0.05）. Compared with the control group， the observation group exhibited reduced PI， RI， LH， T， DHEAS， FINS， FPG， HOMA-IR， TNF-α， hs-CRP， and IL-6 （P<0.01）， but elevated E2， FSH， and APN （P<0.01）. Besides， the bilateral ovarian volume and combined phlegm and stasis-induced syndrome score of the observation group were smaller than those of the control group （P<0.01）， while the endometrial thickness and proportion of type A endometrium were higher （P<0.01）.ConclusionOn the basis of CC treatment， the modified Huanglian Wendantang and Shaofu Zhuyutang alleviates the ovulation disorder in PCOS patients of combined phlegm and stasis-induced syndrome and regulates IR and chronic inflammation， thus creating a favorable condition for clinical pregnancy， which is worthy of further research.
Keywords：polycystic ovary syndrome;chronic inflammation;combined phlegm and stasis-induced;Huanglian Wendantang;Shaofu Zhuyutang;insulin resistance
Abstract：ObjectiveTo study the intervention effect and underlying mechanism of Fufang Huangbaiye Tuji （FFHBY） on skin with deep Ⅱ degree burn wound.MethodPatients with deep Ⅱ degree burn of fire-toxin injuring fluid syndrome diagnosed in the Affiliated Hospital of Chengde Medical University from June 2019 to June 2020 were randomly divided into a control group （iodophor solution， 35 mL per 1% body surface area）， a low-dose treatment group （FFHBY， 17.5 mL per 1% body surface area）， and a high-dose treatment group （FFHBY， 35 mL per 1% body surface area）， 40 cases in each group. The patients in each group were treated correspondingly with dressing chance once per day. The pathological changes of the wound were observed on the 14th day after treatment. Wound symptoms and signs in each group before treatment and on the 7th， 14th， and 21st days after treatment were quantified， and the clinical efficacy on the 21st day after treatment was evaluated. Wound healing rates in each group were calculated on the 7th， 14th， and 21st days after treatment. The levels of vascular endothelial growth factor （VEGF）， fibroblast growth factor （FGF）-2， FGF-7， epidermal growth factor （EGF）， interleukin （IL）-10， tumor necrosis factor （TNF）-α， and Caspase-3 in wound tissues were measured with enzyme-linked immunosorbent assay （ELISA）. Nuclear factor kappa-B （NF-κB） p65 expression in wound surface was detected by immunohistochemistry. The apoptosis rate in wound tissues was determined by the TdT-mediated dUTP-biotin nick end labeding assay （TUNEL） method.ResultThere was no significant difference in scores of symptoms and signs among groups before treatment. Compared with the control group， the treatment groups showed no significant difference in wound healing rates on the 7th day after treatment and increased healing rates on the 14th and 21st day after treatment（P<0.05）. The clinical efficacy in the treatment groups was superior to that in the control group on the 21st day after treatment. Additionally， the treatment groups also showed decreased scores of local symptoms and signs， increased levels of VEGF， FGF-2， FGF-7， EGF， and IL-10， and dwindled apoptosis rate and levels of Caspase-3， TNF-α， and NF-κB p65 expression in wound tissues on the 7th，14th and 21st day after treatment （P<0.05）. The high-dose treatment group was superior to the low-dose treatment group in the above indicators （P<0.05）. Histopathological examination showed that inflammatory cell infiltration was relieved in the treatment groups as compared with that in the control group， and the high-dose treatment group exhibited superior efficacy.ConclusionFFHBY had an obvious therapeutic effect on deep Ⅱ degree burn. It could promote wound healing by up-regulating the level of growth factors， improving inflammatory response， and inhibiting cell apoptosis in a dose-dependent manner.
Abstract：ObjectiveTo evaluate of modified Shentong Zhuyutang combined with celecoxib capsules in the treatment of acute discogenic low back pain due to wind-cold and stasis obstruction sydrome.MethodOne hundred and forty eligible patients were randomly divided into control group （70 cases） and observation group （70 cases）. Patients in both groups took the celecoxib capsules orally， 200 mg/time. On this basis， patients in the control group were further treated with Tongluo Huoxue capsule orally， 6 capsules/time， 3 times/day， while those in the observation group received the modified Shentong Zhuyutang， 1 bag/day， for three weeks. The short-form McGillpain questionnaire （SF-MPQ）， including pain rating index （PRI）， present pain intensity （PPI）， and visual analog scale （VAS）， Oswestry disability index （ODI）， wind-cold obstruction and stasis syndrome， and abilities of daily living （ADL） scoring and the Schober's test were conducted before and after treatment. The tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， thromboxane B2 （TXB2）， and 6-keto prostaglandin F1α （6-keto-PGF1α） levels before and after treatment were measured， followed by the safety evaluation.ResultThe PRI， VAS， and PPI scores in SF-MPQ of the observation group were reduced in contrast to those in the control group （P<0.01）. The wind-cold and stasis obstruction syndrome and ODI scores in the observation group also declined as compared with those of the control group （P<0.01）， while the ADL and Schober's test scores were elevated （P<0.01）. The TNF-α， IL-1β， IL-6， and TXB2 levels in the observation group were lower than those in the control group （P<0.01）， whereas the 6-keto-PGF1α was higher （P<0.01）. The excellent and good rate of the observation group was （51/63） 80.95%， significantly better than （39/63） 61.90% of the control group （χ2＝5.601， P<0.05）. No adverse reactions occurred after the intake of Chinese medicinal prescriptions.ConclusionThe modified Shentong Zhuyutang combined with celecoxib capsules is efficient and safe in inhibiting inflammation， easing back pain， enhancing lumbar spine mobility， and improving the ADL of patients with acute DLBP of wind-cold obstruction syndrome.
Keywords：discogenic low back pain;wind-cold and stasis obstruction syndrome;acute phase;Shentong Zhuyutang;anti-inflammation;analgesia
Abstract：ObjectiveTo systematically analyze the chemical constituents of Qizhi Jiangtang capsules by ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）.MethodAnalysis was conducted on a ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 μm） with acetonitrile （A）-water （B） as the mobile phase for gradient elution （0-13 min， 1%-25%A； 13-21 min， 25%-35%A； 21-28 min， 35%-85%A； 28-30 min， 85%-100%A； 30-32 min， 100%-1%A）. The flow rate was 0.2 mL·min-1， the column temperature was 30 ℃， and the volume of sample injection was 3 μL. Electrospray ionization （ESI） was used to collect data in the negative and positive ion modes with the scanning range of m/z 100-1 500. Meanwhile， a variety of MS analytic methods were used， including comparing with the information of control substances， self-built compounds database and literature references， diagnostic ion filtering， Compound Discoverer 3.0 software， for identification of the chemical components.ResultBased on the above strategy， a total of 52 compounds were identified in Qizhi Jiangtang capsules， and the sources of these compounds were identified. Amino acids were mainly derived from Hirudo， phenylpropanoids were derived from Astragali Radix and Rehmanniae Radix， iridoid glycosides were derived from Rehmanniae Radix， coumarins and triterpenes were derived from Astragali Radix， flavonoids were from Astragali Radix and Polygonati Rhizoma.ConclusionThe established UPLC-QE-Orbitrap-MS analytical method can comprehensively and rapidly analyze and identify of the chemical constituents in Qizhi Jiangtang capsules. Many of the ingredients have been proved by modern pharmacological studies to have the effect of improving related symptoms of diabetes and its complications， reflecting the characteristics of synergistic action of multiple components in Qizhi Jiangtang capsules. This study can provide reference for the further research on the pharmacodynamic material basis and the quality control of Qizhi Jiangtang capsules.
Keywords：Qizhi Jiangtang capsules;ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）;chemical constituents;fragmentation pattern;flavonoids;iridoid glycosides;phenylpropanoids
Abstract：ObjectiveTo explore the mechanism of rancidity during storage by researching the changes of water content， relative permeability of cell membrane and rancidity levels of Armeniacae Semen Amarum in deterioration process.MethodArmeniacae Semen Amarum samples under different storage conditions were evaluated and classified by sensory assessors， and samples with different levels of rancidity were obtained. Water content was measured by toluene method， and water activity was obtained by water activity meter. Malondialdehyde （MDA） and relative conductivity were measured using thiobarbituric acid colorimetry and conductivity meter， respectively. The content of fatty oil was obtained by Soxhlet extraction. The acid value and peroxide value were measured in accordance with the general rules 0713 and 2303 of the 2020 edition of Chinese Pharmacopoeia （part Ⅳ）， respectively. Based on the above experimental data， chemometric methods （cluster analysis， principal component analysis） were selected to establish classification and discriminant models of Armeniacae Semen Amarum with different rancidity levels， in order to verify the accuracy of the classification results.ResultAccording to the results of sensory evaluation， Armeniacae Semen Amarum samples were divided into three classes， including no rancidity， slight rancidity and rancidity. Compared with the no rancid samples， the color of surface and cotyledon were deepened in rancid samples， and the oil was appeared on surface with rancid taste. The values of water content， water activity， MDA content and relative conductivity were all significantly increased in deterioration process （P<0.01）. The content of fatty oil was significantly decreased with the occurrence of rancidness （P<0.01）， while the acid value and peroxide value increased significantly （P<0.01）. The results of cluster analysis and principal component analysis showed that the rancid samples could be distinguished from the no rancid and slightly rancid samples.ConclusionThe storage conditions under high temperature and high humidity can accelerate the rancidness of Armeniacae Semen Amarum， which is accompanied by the increase of internal water content， the increase of cell membrane permeability and the occurrence of fatty acid rancidity. It is suggested that Armeniacae Semen Amarum should be stored in low temperature， dry environment， as well as short storage time.
Abstract：ObjectiveTo explore active components and mechanism of Dipsaci Radix in treating rheumatoid arthritis （RA） based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry （UPLC-QTOF-MS/MS） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）.MethodUPLC-QTOF-MS/MS with electrospray ionization （ESI） was used to qualitatively analyze the chemical components in methanol extract of Dipsaci Radix under positive and negative ion scanning modes. The mobile phase consisted of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-10 min， 0.2%-20%B； 10-20 min， 20%-40%B； 20-25 min， 40%-50%B； 25-33 min， 50%-98%B； 33-35 min， 98%-0.2%B）， and the scanning range was m/z 50-2 000. Based on TCMIP， candidate target groups of Dipsaci Radix， RA and syndrome with Yin deficiency of liver and kidney were obtained， and correlation analysis on "disease-syndrome-prescription" was used to enrich the main active components and key targets. Cytoscape 3.8.0 and STRING 11.0 database were used to construct protein-protein interaction （PPI） network diagram. Metascape platform was used to analysis gene ontology biological progress and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathways.ResultA total of 81 ingredients were identified by UPLC-QTOF-MS/MS. Based on TCMIP， 283 candidate targets corresponding to 81 ingredients， 7 RA related targets and 215 genes corresponding to syndrome with Yin deficiency of liver and kidney were collected. With further correlation analysis on "disease-syndrome-prescription"， 17 key active ingredients were predicted， mainly including saponins and fatty acids of Dipsaci Radix. It mainly involved 7 hub targets， namely tumor necrosis factor （TNF）， nuclear factor-κB subunit 1 （NF-κB1）， hepatocyte nuclear factor 4 alpha （HNF4A）， nuclear receptor subfamily 3 group C member 1 （NR3C1）， peroxisome proliferator activated receptor gamma （PPARG）， nuclear receptor subfamily 1 group H member 4 （NR1H4） and nuclear receptor coactivator 1 （NCOA1）. All of them were related to inflammation， and two of them were related to bile acid pathway. The 7 hub targets and 7 pathways played an important role in RA were screen out， including 4 bile acid related pathways and 3 inflammatory related pathways.ConclusionUPLC-QTOF-MS/MS combined with TCMIP preliminarily elucidates the regulatory effect of multi-components in Dipsaci Radix on several pathways related to the inflammatory response and bile acid synthesis and metabolism， which lays a foundation for further study on the mechanism of Dipsaci Radix against RA.
Abstract：ObjectiveTo study the correlation of eight chemical components in Miao medicine Periploca forrestii from different producing areas with the ecological and soil factors.MethodThe contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， isochlorogenic acid B， isochlorogenic acid A， isochlorogenic acid C， procyanidin A2， and periplocin were simultaneously determined by ultra-performance liquid chromatography （UPLC）. The root soil samples from various producing areas were collected for testing various soil factors， followed by climate data extraction with ArcGIS and topographic data recording using GPS. SPSS 24.0 was employed to conduct the bivariate analysis and stepwise regression analysis of the eight chemical components in P. forrestii from different producing areas with the ecological and soil factors.ResultStepwise regression equations of the content of eight chemical components against ecological and soil factors were established. The findings demonstrated that neochlorogenic acid was negatively correlated with precipitation in the coldest season and chlorogenic acid negatively correlated with precipitation in the driest month. Cryptochlorogenic acid was negatively correlated with precipitation in the coldest season and average temperature in the warmest season， but positively with selenium. Isochlorogenic acid B was mainly affected by soil factors. Specifically， it was positively correlated with available iron and molybdenum but negatively with total phosphorus and available phosphorus. Isochlorogenic acid A was positively correlated with molybdenum but negatively with the coefficient of variation of precipitation. Isochlorogenic acid C showed a positive correlation with exchangeable magnesium. Procyanidin A2 exhibited a positive correlation with molybdenum and a negative correlation with available potassium. Periplocin was negatively correlated with the coefficient of variation of precipitation.ConclusionThe correlation between the eight chemical components of P. forrestii and the ecological and soil factors has been clarified， which will provide reference for the introduction， cultivation， and standardized planting of P. forrestii and also a theoretical basis for further research on its ecological and soil factors and quality formation mechanism.
Abstract：ObjectiveTo identify 24 Rana species such as Rana dybowskii by mitochondrial cytochrome C oxidase subunit I （COⅠ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of Rana species as well as the discovery of new species.MethodR. dybowskii， R. chensinensis， R. amurensis， R. culaiensis， and R. huanrenesis， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification and sequencing. A total of 50 COⅠ gene sequences were obtained. Then 163 COⅠ gene sequences for 24 species of Rana and one COⅠ gene sequence for Pelophylax， Odorrana， Nidirana， Hylarana， and Amolops were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of COⅠ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis.ResultThe COⅠ gene sequences of 24 Rana species including R. dybowskii were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between R. sangzhiensis and R. zhengi was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. R. sangzhiensis and R. zhengi were clustered into one branch and some R. dybowskii and R. uenoi into one branch. There were two separate branches for R. chensinensis and the other species were all clustered independently.ConclusionCOⅠ-based DNA barcoding enabled the identification of 24 species of Rana including R.dybowskii. The findings supported that R. sangzhiensis， R. zhengi， R. coreana， and R. kunyuensis were the same species. One branch of R. chensinensis might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that COⅠ-based DNA barcoding allows not only the identification of 24 species of Rana including R. dybowskii but also the classification of ranidae species and the discovery of new species or subspecies.
Abstract：ObjectiveDNA barcodes suitable for Lauraceae plants were screened，and 22 Lauraceae plants were identified and classified.MethodThe DNA of 22 species of Lauraceae was extracted and amplified by polymerase chain reaction（PCR） with different DNA barcoding primers，followed by electrophoresis and sequencing. Codon Code Aligner was used to proofread，splice， and remove the forward and reverse primer sequences. The sequence was imported into DNAMAN for multiple sequence alignment，and the base mutation sites were analyzed. The Kimura 2-Parameter（K2P） distance of different plants was calculated by MEGA，and the variation degree was analyzed according to the genetic distance. The phylogenetic tree was constructed based on the adjacency method.ResultThree pairs of DNA barcoding primers were used to amplify and sequence the DNA of 22 species of Lauraceae，and 20 species were identified by comparing the specific base sites of gene sequences.ConclusionFour Litsea plants could be identified by matK， three Phoebe plants except for Cinnamomum validinerve by rbcL， and 20 Lauraceae plants by the combination of matK， rbcL， and rpoB，which provided a theoretical basis for the identification and development of Lauraceae plants. Among them，matK was predominant in the identification of Lauraceae plants，and the results also showed that the combination of sequences for plant identification could achieve a better result in DNA barcoding. This study investigated the genetic relationship between Lauraceae plants at the molecular level，aiming at providing a basis for the investigation，cultivation，development，protection， and utilization of medicinal plant resources.
Abstract：ObjectiveGuided by nutrient-defense stages in the vessel-collateral theory， the modern medical cases of unstable angina pectoris（UAP） were systematically collated and analyzed to explore the rules of syndrome and treatment of UAP and the molecular mechanism of core Chinese herbal combination in the treatment of UAP based on network pharmacology.MethodAll medical cases with UAP treated by Chinese medicinal compounds were retrieved from PubMed，China National Knowledge Infrastructure （CNKI），Wanfang Data， VIP， and SinoMed published between database inception and November 2020. The syndromes of medical cases were determined based on the nutrient-defense stages of the vessel-collateral theory. Rules of syndrome and treatment of UAP were investigated by data mining methods， such as frequency statistics， cluster analysis， and enhanced FP-Growth algorithm. The molecular mechanism of core Chinese herbal combination in the treatment of UAP was analyzed by network pharmacology.ResultThe first four syndromes of UAP with high frequencies were deficiency and stagnation of collateral Qi， blood stasis obstructing collaterals， depression and stagnation of collateral Qi， and turbid phlegm obstructing collaterals. The Chinese herbal medicines with high frequencies included Salviae Miltiorrhizae Radix et Rhizoma， Glycyrrhizae Radix et Rhizoma， Chuanxiong Rhizoma， and Astragali Radix， which were effective in resolving stasis， dredging collaterals， replenishing Qi， consolidating defensive Qi， regulating Qi， relieving depression， and dispelling phlegm. The association analysis indicated that the core Chinese herbal combination in the treatment of UAP was Salviae Miltiorrhizae Radix Chuanxiong Rhizoma-Astragali Radix. Four Chinese herbal combinations were obtained by cluster analysis. As revealed by network pharmacology， the key components of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma-Astragali Radix in the treatment of UAP included quercetin， luteolin， and tanshinone ⅡA， and the key targets included serine/threonine-protein kinase 1 （Akt1）， mitogen-activated protein kinase 1 （MAPK1）， Jun， interleukin （IL）-6， and MAPK8. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis showed that phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathway might serve as the main pathway for its therapeutic efficacy.ConclusionThe basic pathogenesis of UAP is deficiency/depression and stagnation of collateral qi and turbid phlegm obstructing collaterals. The treatment should follow the principles of replenishing Qi， resolving stasis， and dredging collaterals， assisted with regulating Qi and resolving phlegm. The therapeutic efficacy of Salviae Miltiorrhizae Radix-Chuanxiong Rhizoma-Astragali Radix was achieved via multi-component， multi-target， and multi-pathway. This study is expected to inspire future UAP-related studies at the molecular level based on vessel-collateral theory.
Keywords：nutrient-defense stages of vessel-collateral theory;unstable angina pectoris （UAP）;data mining;network pharmacology;rules of syndrome and treatment
Abstract：ObjectiveTo study the material basis and potential molecular mechanism of Epimedii Folium against osteoporosis.MethodThe chemical components in 14 batches of Epimedii Folium were analyzed by ultra-performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry （UPLC-Q-TOF-MS/MS）. With the activity of alkaline phosphatase （ALP） as the pharmacodynamic index，the partial least squares regression analysis （PLSR） was conducted to establish a model uncovering the spectrum-effect relationship between UPLC-Q-TOF-MS/MS spectral peak and ALP activity and screen the active components against osteoporosis. Online databases such as the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP），Comparative Toxicogenomics Database （CTD），Database for Annotation，Visualization，and Integrated Discovery （DAVID） and Cytoscape 3.6.1 were employed to predict the possible mechanism of action of Epimedii Folium against osteoporosis.ResultA total of 61 peaks and 56 compounds were identified by UPLC-Q-TOF-MS/MS. PLSR showed that icariin，baohuoside Ⅰ，epimedin A，sagittatoside A，and baohuoside Ⅱ might be the active components for Epimedii Folium to inhibit osteoporosis considering their strong correlation with ALP activity. As revealed by the network pharmacological analysis of the five components mentioned above，Epimedii Folium mainly regulated seven targets such as tumor necrosis factor （TNF），androgen receptor （AR），and estrogen receptor 1 （ESR1） and eight key pathways like endocrine and other factor-regulated calcium reabsorption，vascular endothelial growth factor （VEGF） signaling pathway，and transient receptor potential （TRP） channels to exert its anti-osteoporosis effect.ConclusionThe exploration of material basis and potential molecular mechanism of Epimedii Folium against osteoporosis based on UPLC-Q-TOF-MS/MS，spectrum-effect relationship，and network pharmacology has provided an experimental basis for the scientific explanation and clinical application of Epimedii Folium in treating osteoporosis.
Keywords：Epimedii Folium;ultra-performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry（UPLC-Q-TOF-MS/MS）;anti-osteoporosis;spectrum-effect relationship;network pharmacology
Abstract：Traditional Chinese medicine （TCM） is a great treasure house， exhibiting unique advantages in the treatment of some difficult and critical diseases. The incidence rate of membranous nephropathy has increased year by year in recent years， and has become the first cause of primary glomerular diseases. However， its pathogenesis is not clear. Modern medicine often uses immunosuppressive therapy， but it often faces the problems of high side effects and high recurrence rate. The China Association of Chinese Medicine （CACM） invited clinical experts of TCM and western medicine to fully discuss membranous nephropathy， which was later confirmed to be one of the clinical diseases responding specifically to TCM. Apart from summarizing the pathogenesis and clinical diagnosis and treatment of membranous nephropathy in both TCM and western medicine， this paper also detailed TCM cognition， syndrome differentiation， and therapeutic schemes of membranous nephropathy， aiming to improve the clinical remission rate of membranous nephropathy and provide reference for its clinical treatment.
Keywords：nephrology department;membranous nephropathy;integrated traditional Chinese medicine （TCM） and western medicine;diseases responding specifically to TCM;expert consensus
Abstract：Zexietang is derived from Jingui Yaolue （《金匮要略》）， which is composed of Alismatis Rhizoma and Atractylodis Macrocephalae Rhizoma， and has the effect of inducing diuresis and invigorating the spleen to produce water. Compared with western medicine in the treatment of related diseases， Zexietang can not only improve the curative effect， but also reduce the occurrence of adverse reactions， so as to achieve long-term stable administration. The authors sorted out and analyzed the chemical composition， pharmacological effect and clinical application of Zexietang in recent years. It was found that the main active components of Zexietang were alismol A and B， 23-acetyl-alismol B and C， atractylenolides （atractylenolide Ⅰ， Ⅱ， Ⅲ） and polysaccharides. Pharmacological experiments showed that they had diuretic， hypolipidemic， anti-inflammatory and others. And it can be used in the treatment of hypertension， hyperlipidemia， vertigo， cerebral vascular insufficiency and other diseases combined with other Chinese materia medica， and the curative effect is obvious. By summarizing the research status of Zexietang in recent years， its active components and pharmacological mechanism can be further clarified， which provides the basis for the clinical application of Zexietang and guides the direction of its further research.
Abstract：The Leguminosae perennial vines of Callerya and Millettia have many species and wide distribution， not only can be used for medicines， but also they have ornamental and insecticidal effects. With increasing demand for Spatholobi Caulis， and the reserves of wild medicinal materials are on the verge of exhaustion， resulting in the increasing number of mixtures and substitutes in the market， which makes it urgent to study the origin of Spatholobi Caulis. By referring to related literature， there are three major origins of Spatholobi Caulis， including Callerya， Millettia and Spatholobus. Callerya is separated from Millettia， they are divided and united for many times， now the official website of Flora of China has accepted the revision of them as two genera. This paper intends to compare the chemical components and pharmacodynamic effects of Callerya and Millettia， aiming to explore the similarities and differences between the two genera， so as to determine the rationality and necessity of separating Callerya from Millettia. After comparing， it was found that the chemical composition and pharmacodynamic effects of the two genera were different， which supported the separation of Callerya from Millettia， and it was not recommended to mix use of them.
Keywords：Callerya;Millettia;historical evolution;molecular identification;chemical constituents;pharmacological activity;quality control
Abstract：Diarrhea is characterized by increased bowel movements and loose and even watery stools. Its occurrence and development have been proved by modern research to be closely related to the imbalance of intestinal flora. Traditional Chinese medicine （TCM） pays a special attention to syndrome differentiation in treating diarrhea. Exploring the TCM syndromes of diarrhea is of great significance to the formulation of TCM diagnosis and treatment scheme of diarrhea and the improvement of clinical curative effects. There exist many similarities between TCM theory and micro-ecological theory concerning diarrhea. With the deepening of intestinal flora research， the significance of intestinal flora in TCM syndrome research has been increasingly highlighted. The close correlation of intestinal flora with the occurrence and development of diarrhea has provided new ideas of deducing syndrome and selecting prescription based on intestinal flora. This paper summarized the relationship between TCM syndromes of diarrhea and microscopic indexes such as immune response， neurotransmitters， brain-gut peptide， and proteins and analyzed the intestinal flora characteristics related to six common TCM syndromes of diarrhea. Meanwhile， based on the theory of deducing syndrome by prescription， namely deducing the syndrome by the efficacy of prescription or its medicinal components， the correlation between TCM syndromes of diarrhea and intestinal flora was indirectly verified， so as to identify the research direction of correlation between intestinal flora and TCM syndromes in the future. Numerous studies have shown that the TCM syndromes of diarrhea were highly correlated with the microscopic indexes such as inflammatory cytokines， neurotransmitters， and proteins. Syndromes and dysbacteriosis both resulted from pathogenic factors acting on the body， which were summarized from different angles. Different TCM syndromes corresponded to specific objective indicators of intestinal flora. Intestinal flora has the potential of being an internal material basis for powerfully revealing the TCM syndromes of diarrhea in the future.
Abstract：Depression is a mental illness characterized by persistent negative feelings， which has seriously threatened people's health. In recent years， neuronal autophagy， an important stress response， has also been regarded as a hypothesis for the pathogenesis of depression. Relevant studies have shown that either insufficient or excessive autophagy triggers neuronal damage， and activated or inhibited neuronal autophagy can be observed in animal models of depression. Therefore， neuronal autophagy may be a double-edged sword involved in the pathogenesis of depression. It is believed in traditional Chinese medicine （TCM） that the occurrence of this disease is closely related to liver depression and spleen deficiency. Chinese medicine regulates the neuronal autophagy via multiple ways. The TCM monomers that regulate neuron autophagy are capable of protecting nerves or penetrating the blood-brain barrier. TCM compounds designed for soothing liver or invigorating spleen have been proved effective against this disease， demonstrating that the core pathogenesis of depression lies in liver depression and spleen deficiency. The regulatory effects of TCM on neuronal autophagy in depression models might result from its action on multiple targets， multiple pathways， and multiple systems. This paper discussed the limitations in current research based on the involvement of neuronal autophagy in depression and its treatments， in order to provide ideas for later similar research and that concerning TCM treatment of depression.
Keywords：depression;neuronal autophagy;pathogenesis;therapeutic mechanism;traditional Chinese medicine （TCM） treatment
Abstract：Alzheimer's disease （AD） is currently the most common neurodegenerative disorder characterized by insidious onset and slow progression， which has seriously endangered the physical and mental health of the elderly. It is therefore very important to carry out the early diagnosis and active prevention and treatment. Biomarkers are essential for its diagnosis. Looking for ideal biomarkers is helpful for early identification of this disease. The prevention and treatment of AD with traditional Chinese medicine （TCM） has always been a hot spot in scientific research due to good safety and small side effects. Proteomics is an advanced omics technology that enables the identification and characterization of proteins. Its high-throughput， dynamic， and comprehensive characteristics coincide with the concept of holism emphasized in TCM diagnosis and treatment， which makes proteomics suitable for identifying biomarkers with diagnostic potential， objectifying TCM syndrome differentiation and treatment， and developing new Chinese medicinal prescriptions for precise and targeted treatment. Although the proteomics technology is becoming increasingly mature， it still faces challenges in the diagnosis and treatment of AD. There exist such shortcomings as high heterogeneity and poor reproducibility of protein omics results， requirement for the combination with other advanced omics technologies， and high sequencing cost. In the future， the protein omics technology should be constantly updated and optimized to boost precision medicine， disease prevention， and drug research and development. This paper retrieved related articles from Pubmed， China National Knowledge Infrastructure （CNKI） and reviewed the application of proteomics in the early diagnosis of AD and its prevention and treatment with TCM in recent years， in order to provide reference for the in-depth study of AD diagnosis as well as its prevention and treatment with TCM.
Abstract：Dahuang Zhechongwan （DHZCW） is a classic prescription from the Jingui Yaolue（《金匮要略》） by ZHANG Zhong-jing，with the effects of tonifying deficiency， relaxing the middle， promoting regeneration， and resolving stasis. It has been widely used in the clinical treatment of various diseases with definite efficacy achieved. The research on multiple organ fibrosis has shown that DHZCW can slow down the development of organ fibrosis in the heart， liver， kidney， lung， etc.， and good results in both clinical practice and experimental research have been obtained. The present study reviewed the previous investigations on the experimental mechanism of DHZCW in the treatment of multiple organ fibrosis and revealed that the pathogenesis was closely related despite different disease sites. From the perspective of traditional Chinese medicine （TCM），these diseases shared a common pathogenesis，which was manifested by deficiency. Long-term diseases led to the formation of "dried blood". From the perspective of modern medicine， the diseases all showed pathological changes in the deposition of extracellular matrix （ECM）， and their occurrence and development were all based on certain effector cells ［such as hepatic stellate cell （HSC） and pancreatic stellate cell （PSC）］， with same cytokines ［such as tumor necrosis factor-α （TNF-α），interleukin-6 （IL-6），IL-1β，and α-smooth muscle actin （α-SMA）］ and some key pathways ［transforming growth factor-β1 （TGF-β1）/Smad， phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt）， and lipopolysaccharide （LPS） paracrine and autocrine mechanisms］ involved. As a classic prescription for "deficiency-induced dry blood"， DHZCW was effective in treating fibrosis， which was presumedly related to the inhibition of ECM deposition by intervening in the above-mentioned mechanisms， thereby delaying the disease progression. This study is expected to provide literature support to clarify the scientific connotation of DHZCW in the treatment of multiple organ fibrosis and lay a foundation for further experimental and clinical research.
Keywords：Dahuang Zhechongwan;multiple organ fibrosis;experimental research;review
Abstract：At present， the incidence and mortality of gastric cancer in China are 10.26% and 12.45%， ranking the second and third places， respectively， in the incidence and mortality of malignant tumors in China. Cancer often goes through three stages： precancerous lesions， carcinoma in situ， and invasive carcinoma. It is of great significance to advance cancer prevention and control to the stage of precancerous lesions. The popularization of digestive endoscopy-assisted diagnosis and treatment has enabled the timely diagnosis and treatment of early gastric cancer and severe dysplasia. However， the endoscopic mucosal resection is not suitable for most precancerous lesions of gastric cancer （PLGC）， and the effective drugs are not available. The long-term clinical dynamic monitoring has imposed considerable physical and mental burdens on patients. Gastrointestinal microenvironment is a dynamic balance system composed of gastrointestinal flora， chemical barrier， mechanical barrier， immune barrier， and gastrointestinal nerves. The imbalance of gastric microenvironment has been proved to be the key mechanism of PLGC. According to traditional Chinese medicine （TCM）， PLGC is a result of long-term interaction between deficient healthy Qi and excessive pathogens. In syndrome differentiation and treatment， PLGC is often believed to be developed from chronic gastritis. Besides， the inflammation-cancer transformation model put forward by Correa and the evolution of its TCM pathogenesis are also considered. Guided by the basic treatment principle of reinforcing healthy Qi to eliminate pathogenic factors， we determined the basic therapeutic methods as follows： invigorating spleen， clearing heat， and resolving dampness. At the same time， such methods as soothing liver and regulating Qi， resolving phlegm and dissipating mass， activating blood and resolving stasis， clearing heat and removing toxin， and tonifying deficiency can be combined based on the results of syndrome differentiation. After discussing the correlation between the imbalance of gastric microenvironment and PLGC and summarizing TCM intervention methods and mechanisms against PLGC from the perspective of gastric microenvironment regulation， this paper believed that TCM improved the gastric microenvironment by regulating the disorder of gastric flora， eliminating the gastric mucosal inflammation， and relieving the abnormal immune response， thereby preventing and controlling the PLGC.
Keywords：gastric microenvironment;precancerous lesions of gastric cancer （PLGC）;traditional Chinese medicine （TCM） therapies