Abstract：ObjectiveTo rapidly identify the chemical constituents in Xiao Chengqitang by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）.MethodThe method was established by the Waters CORTECS T3 column （2.1 mm×150 mm， 1.6 μm）， mobile phase was methanol （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-5 min， 3%-21%A； 5-20 min， 21%-36%A； 20-32 min， 36%-50%A； 32-42 min， 50%-62%A； 42-50 min， 62%-85%A； 50-60 min， 85%-95%A）， the flow rate was 0.2 mL·min-1， and the column temperature was 30 ℃. UPLC-Q-Orbitrap-MS was operated in positive and negative ion modes， the scanning range was 100-1 200 with mode of Full MS/dd-MS2， and the collision energies were 20， 40 eV. The compounds were identified by comparing with reference substances and combining with literature reports and MS database information.ResultA total of 123 components were identified in Xiao Chengqitang， including 33 flavonoids， 25 anthraquinones and anthrones， 23 phenylpropanoids， 15 tannins， 10 nitrogen-containing components and 17 other components. Among them， 32 components were determined by reference substances.ConclusionThe material basis of Xiao Chengqitang is flavonoids， anthraquinones and anthrones， phenylpropanoids， which is derived from Aurantii Fructus Immaturus， Rhei Radix et Rhizoma and Magnoliae Officinalis Cortex， respectively.
Keywords：Xiao Chengqitang;ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）;famous classical formulas;anthraquinones;anthrones;flavonoids;phenylpropanoids
Abstract：ObjectiveTo investigate the therapeutic effect of Guizhi Shaoyao Zhimutang （GSZT） on the osteoporosis （OP） in SD female rats induced by ovariectomy （OVX） combined with glucocorticoid injection and its related mechanisms.MethodThe rats were divided into a sham operation group and an experimental group for OVX. One week later， the experimental rats were divided into an OP model group， a calciferol （positive control， 0.1 mg·kg-1） group， and low（0.8 g·kg-1）， medium（1.6 g·kg-1）， and high-dose （3.2 g·kg-1） GSZT groups. Except for those in the sham operation group， the rats received an intramuscular injection of dexamethasone （1 mg·kg-1） twice per week for six weeks. After the rats were treated correspondingly for eight weeks， the rats were sacrificed for thighbone sample collection. The computer microtomography （Micro CT） was used to analyze the parameters of bone mineral density （BMD）， bone volume （BV）， tissue volume （TV）， structure model index （SMI）， connectivity density （Conn.D）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， trabecular pattern factor （Tb.Pf）， and trabecular separation （Tb.Sp）. In addition， hematoxylin-eosin （HE） and Masson staining was used for the histopathological examination of the thighbone. Tartrate-resistant acid phosphatase （TRAP） staining was used to analyze osteoclasts in bone tissues. The enzyme-linked immunosorbent assay （ELISA） was used to determine matrix metalloproteinase-9 （MMP-9）， cathepsin K （CTSK）， and TRAP5b in bone tissues， and Western blot was carried out to determine receptor activator of NF-kappaB ligand （RANKL）， osteoprotegerin （OPG）， alkaline phosphatase （ALP） in bone tissues.ResultCompared with normal group， the number of bone trabeculae in cancellous part of OP model rats decreased significantly， the spacing widened， the thickness became thinner， and the continuity was poor. Compared with model group， GZST medium and high-dose groups could improve the changes of femoral bone trabecular cancellous bone. Masson staining showed that compared with the normal group， the growth plate thickness of the distal femur epiphysis was thinner and the area of new bone was smaller in the OP model group， while the reduction of new bone was inhibited in the high dose group of calcitol and GZST. TRAP staining results showed that the number of osteoclasts in OP group was significantly increased compared with the normal group， and the number of osteoclasts in medium-high dose GZST group was reduced. Micro CT results showed that compared with normal group， BMD， BV， Tb.N， Tb.Th and Conn.D of OP rats were decreased， SMI and Tb.Sp were significantly increased（P<0.01）. Compared with model group， medium and high dose of GZST could improve the degree of osteoporosis in OP rats， BV， Tb.N were significantly increased， SMI， Tb.Sp were significantly decreased （P<0.01）. ELISA results revealed that compared with the sham operation group， the OP model group showed decreased ALP content （P<0.01） and increased MMP-9， TRAP5b， and CTSK （P<0.01）. Compared with the OP model group， the high-dose GSZT group showed increased ALP content （P<0.01） and decreased MMP-9， TRAP5b， and CTSK （P<0.01）. Western blot results demonstrated that compared with the OP model group， the calciferol group and the high-dose GSZT group showed inhibited expression of the RANKL protein （P<0.05）， and increased OPG expression （P<0.05）.ConclusionThe findings suggest that GZST has a potential therapeutic effect on OP caused by OVX combined with glucocorticoids in rats， and the underlying mechanism is associated with the regulation of RANKL/OPG in bone tissues. This study is expected to lay a preliminary foundation for its clinical application and modern preparation development.
Abstract：ObjectiveTo explore the underlying mechanism of volatile oil from Sishenwan in treating chronic ulcerative colitis through the Toll-like receptor （TLR）/myeloid differentiation factor 88 （MyD88） signaling pathway.MethodThe BALB/c mice were randomly divided into a normal group （normal）， a model group ［dextran sodium sulfate （DSS）］， a Sishenwan volatile oil group， an Ershen pill volatile oil group， a Wuweizi powder volatile oil group， and a mesalazine control group. The chronic ulcerative colitis model was induced by DSS in mice. Seven days after intragastric administration， the efficacy was evaluated based on the body weight， colon weight， colon weight index， colon length， and pathological damage score under colonoscopy. The levels of interleukin （IL）-4， IL-10， IL-17A， IL-21， and interferon-γ （IFN-γ） in the supernatant of colon tissues were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of proteins related to the TLR/MyD88 signaling pathway in the colon mucosa of mice， including TLR2， MyD88， Ras-related C3 botulinum toxin substrate 1 （Rac1）， IL-1 receptor-associated kinase 4 （IRAK4）， IRAK1， tumor necrosis factor receptor （TNFR）-associated factor 6 （TRAF6）， transforming growth factor-β-activated kinase 1 binding protein 1 （TAB1）， TAB2， mitogen-activated protein kinase kinase 6 （MKK6）， p38 mitogen-activated protein kinase （p38 MAPK）， and cyclic adenosine monophosphate response element-binding protein （CREB）.ResultCompared with the normal group， the model group showed decreased colon length， increased colon weight， colon weight index， and pathological damage score under colonoscopy， decreased IL-10 level in the colon tissues， increased IL-4， IL-17A， IL-21， and IFN-γ levels （P<0.05， P<0.01）， and up-regulated protein expression of TLR2， MyD88， Rac1， IRAK4， IRAK1， TRAF6， TAB1， TAB2， MKK6， p38MAPK， and CREB （P<0.01）. Compared with the model group， the Sishenwan volatile oil group showed increased colon length， reduced colon weight， colon weight index， and pathological damage score under colonoscopy， elevated IL-10 level in the colon tissues， decreased IL-4， IL-17A， IL-21， and IFN-γ levels （P<0.05， P<0.01），and down-regulated protein expression of TLR2， MyD88， Rac1， IRAK4， IRAK1， TRAF6， TAB1， TAB2， MKK6， p38MAPK， and CREB （P<0.05， P<0.01）.ConclusionThe volatile oil from Sishenwan can effectively improve the inflammatory response of chronic ulcerative colitis， which may be achieved by regulating the TLR/MyD88 signaling pathway.
Abstract：ObjectiveTo observe the effects of Chaihu Shugansan on gastric motility and mitochondrial function and mitophagy in rats with functional dyspepsia （FD） and to preliminarily reveal its mechanism in preventing and treating FD.MethodThirty-two SD rats of SPF grade were randomly divided into the normal group， model group， Chaihu Shugansan （4.8 g·kg-1） group， and domperidone group （4.5 mg·kg-1） after one week of adaptive feeding， with eight rats in each group. Rats in all groups except for the normal group were exposed to modified tail-clamping stimulation for inducing FD. Four weeks later， the semi-solid nutritive medium was used to observe the gastric emptying rate of FD rats， and the serum citrate synthase （CS）， motilin （MTL）， and gastrin （GAS） were detected by enzyme-linked immunosorbent assay （ELISA）. The pathological changes in gastric tissue were observed by hematoxylin-eosin （HE） staining， the mitochondrial characteristics by transmission electron microscopy， and the expression of microtubule-associated protein light chain 3 （LC3） and voltage-dependent anion-selective channel 1 （VDAC1） by immunofluorescence co-localization. The mitochondria were extracted from fresh gastric tissue for measuring the reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels using the biochemical kits. The mitochondrial LC3， yeast Atg6 homologous（Beclin1）， and p62 protein expression was assayed by Western blot.ResultCompared with the normal group， the model group exhibited significantly reduced gastric emptying rate （P<0.01） and serum CS， MTL， and GAS levels （P<0.01）. HE staining showed no pathological changes like erosion or ulcer in the gastric tissue of rats in each group. However， the mitochondria in the gastric tissue were observed swollen and dilated under the transmission electron microscope， and vacuolar lesions appeared. The co-expression of LC3 with VDAC1 increased significantly （P<0.01）. The mitochondrial ROS and MDA levels were elevated significantly （P<0.01）， while SOD content was significantly reduced （P<0.01）. The protein expression levels of LC3 and Beclin1 were significantly up-regulated （P<0.05， P<0.01）， whereas p62 protein expression was down-regulated significantly （P<0.01）. Compared with the model group， Chaihu Shugansan and domperidone elevated the gastric emptying rate （P<0.05） and serum CS， MTL， and GAS levels （P<0.05， P<0.01）. It was observed under the transmission electron microscope that the mitochondrial nuclear membrane of the gastric tissue was intact， with clear structure and high density of mitochondrial cristae， and mitochondrial fission and fusion were observed in some parts. The co-localization area of LC3 and VDAC1 was significantly reduced （P<0.01）. The mitochondrial ROS and MDA levels declined significantly （P<0.05， P<0.01）， whereas SOD level rose （P<0.05）. The protein expression levels of LC3 and Beclin1 were significantly down-regulated （P<0.05， P<0.01）， while that of p62 was obviously up-regulated （P<0.01）.ConclusionThe mechanism of Chaihu Shugansan in preventing and treating FD may be related to the improvement of mitochondrial function and the inhibition of mitophagy in gastric tissue.
Abstract：ObjectiveTo seed for stable time window of the integrated disease-syndrome animal model based on the counterevidence from Chinese medicinal prescriptions， and to verify syndrome stability and reliability.MethodA model of depression was established by exposing rats to chronic unpredictable mild stress （CUMS）， followed by body weight measurement， sugar water test， behavioral test， and brain 5-hydroxytryptamine（5-HT） detection. The identification of liver depression and spleen deficiency syndrome was conducted after the equivalent transformation of human clinical symptoms into macroscopic representations of rats. Based on the dynamically collected macroscopic representation scale， Xiaoyaosan was used to reversely verify the stability and reliability of the integrated disease-syndrome animal model of depression due to liver depression and spleen deficiency.ResultThe sugar water consumption and the number of crossings and the total movement distance in the open field test of 16-week-old rats in the CUMS （eight weeks of CUMS） group were significantly lower than those in the normal group （P<0.05）. According to the immunohistochemical results， the 5-HT content in hippocampal area CA2 of rats in the CUMS group was also significantly lowered as compared with that in the normal group（P<0.05），which indicated that depression was successfully modeled. The liver depression and spleen deficiency syndrome was present in 14-week-old rats （six weeks after CUMS）of the CUMS group， and the number of rats experiencing the liver depression and spleen deficiency syndrome reached the peak in the 16th week （eight weeks after CUMS），accounting for 70% of the total number. Thereafter， the number decreased gradually. The syndrome scores of the 14-， 16-， 18-， 20-， and 22-week-old rats in the Xiaoyaosan group were reduced by 66.6%， 70.7%， 54.8%， 50.4%， and 44.8%， which were graded as effective， marked effective， effective， effective， and effective， respectively.ConclusionThe age of 14-16 weeks（six to eight weeks after CUMS） is considered the stable and reliable time window for depression due to liver depression and spleen deficiency.
Keywords：counterevidence from Chinese medicinal prescription;integrated disease-syndrome;animal model;depression;liver depression and spleen deficiency
Abstract：ObjectiveTo investigate the effect of Astragalus polysaccharide （APS） on transforming growth factor-β1 （TGF-β1）-induced epithelial mesenchymal transition （EMT） of A549/DDP lung adenocarcinoma xenograft and its potential molecular mechanism.MethodBALB/c nude mice were randomly divided into the non-loading group （A549/DDP cells not loaded with TGF-β1）， model group， cisplatin group， and combined group （A549/DDP cells overexpressing TGF-β1）. Mice in the combined group were treated with intragastric administration of APS （0.3 g·kg-1·d-1） and intraperitoneal injection of cisplatin （0.003 5 g·kg-1）， while those in the cisplatin group only received intraperitoneal injection of cisplatin （0.003 5 g·kg-1）. After drug intervention， the nude mice were sacrificed and the xenograft and lung were harvested， followed by the weighing of tumor and the calculation of the inhibition rate. The number of tumors metastasizing to the lung was counted under the microscope. The pathological features of tumors and their metastasis to the lung tumor were observed by hematoxylin-eosin （HE） staining. The protein and mRNA expression levels of EMT molecular markers E-cadherin， Vimentin， α-smooth muscle actin （α-SMA）， and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） in the xenograft were detected by immunohistochemistry， Western blot， and Real-time polymerase chain reaction （Real-time PCR）.ResultCompared with the non-loading group， the model group exhibited increased tumor weight and pulmonary metastatic nodules （P<0.05）， sparse tumor cell junctions， long spindle cells， massive metastatic nodules in the lung， down-regulated E-cadherin protein and mRNA expression， and up-regulated Vimentin and α-SMA protein and mRNA expression and p-PI3K and p-Akt protein expression （P<0.05）. Compared with the model group and cisplatin group， the combined group displayed decreased tumor weight and pulmonary metastatic nodules （P<0.05）， tight tumor cell junctions， round or oval cells， no obvious lung metastasis， up-regulated E-cadherin protein and mRNA expression （P<0.05）， and down-regulated Vimentin and α-SMA protein and mRNA expression （P<0.05） and p-PI3K and p-Akt protein expression （P<0.05）. There was no significant difference in PI3K or Akt protein expression among groups.ConclusionAPS has a certain inhibitory effect against EMT in lung adenocarcinoma A549/DDP cells， which may be related to the inhibition of activated PI3K/Akt protein expression.
Abstract：ObjectiveTo investigate the inhibitory effects of Danshen injection against ovarian cancer cell proliferation induced by the interaction between platelets and cancer cells.MethodThe induction of platelets on SKOV3 growth in vitro and the inhibitory effect of Danshen injection at 12，24，and 48 g·L-1 were observed by 3-（4，5-Dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） and cell colony formation assays. The content of transforming growth factor-β1 （TGF-β1） in the platelet-tumor cell interaction system and platelet supernatant and the effect of Danshen injection on TGF-β1 secretion were detected by enzyme-linked immunosorbent assay （ELISA）. The influences of tumor cell culture supernatant on platelet aggregation and secretion and the inhibitory effect of Danshen injection were determined by microplate assay and ELISA. The effects of Danshen injection on platelet nuclear factor kappa B （NF-κB） signaling pathway were assayed by Western Blot.ResultCompared with the blank group， the platelet induction group exhibited significantly elevated absorbance at A570 （P<0.01）， while the absorbance at A570 in the platelet + Danshen injection group was significantly lower than that in the platelet induction group （P<0.01）. The comparison with the Danshen injection group revealed that the cell proliferation inhibitory rate in the platelet + Danshen injection group at the same dose was more significant （P<0.01）. The number of colonies in the platelet induction group was obviously increased in contrast to that in the blank group（P<0.05）， while the number of colonies in the platelet + Danshen injection group was significantly lower than that in the platelet induction group（P<0.05，P<0.01）. As demonstrated by comparison with the blank group， TGF-β1 content in the supernatant of the platelet induction group rose remarkably（P<0.01）， whereas that in the platelet + Danshen injection group declined（P<0.01）. Compared with the Danshen injection （24 g·L-1） group， the platelet + Danshen injection group displayed more obvious inhibition（P<0.01）. Compared with the blank group， Danshen injection significantly reduced the TGF-β1 content in platelet supernatant（P<0.05，P<0.01）. There was no significant change in the content of TGF-β1 in SKOV3 supernatant treated with Danshen injection. The platelet aggregation， thromboxane A2（TXB2）， and serotonin （5-HT） secretion in the SKOV3 cell supernatant induction group were significantly increased as compared with those in the blank group （P<0.01）， while such indexes in the cell supernatant induction + Danshen injection group were obviously decreased （P<0.01）. Compared with the Danshen injection （24 g·L-1） group， the cell supernatant induction + Danshen injection group displayed more obvious inhibition at the same dose（P<0.01）. Compared with the blank group， the platelet induction group exhibited obviously up-regulated phosphorylated TGF-β-activated kinase-1 （TAK-1） and NF-κB， but down-regulated phosphorylated inhibitory protein of NF-κB （IκB）（P<0.01）， which however were significantly reversed in the platelet + Danshen injection group（P<0.01）.ConclusionDanshen injection affect the proliferation of SKOV3 cells by inhibiting their interaction with platelets， which may be related to the inhibited secretion of TGF-β1.
Abstract：ObjectiveTo observe the effect of Jianpi Xiaoai prescription on long non-coding RNA Hox transcript antisense intergenic RNA （lncRNA HOTAIR）/Janus kinase 2 （JAK2） /signal transducer and activator of transcription 3 （STAT3） signaling pathway and to explore the potential mechanism of Jianpi Xiaoai prescription in suppressing the metastasis of colon cancer.MethodThe expression of lncRNA HOTAIR in different cells was analyzed. Following the treatment of HCT116 cells with 10%，15%，and 20% Jianpi Xiaoai prescription -containing serum， the invasive ability of Jianpi Xiaoai prescription on HCT116 cells was assessed by transwell assay. The mRNA expression levels of lncRNA HOTAIR，JAK2，and STAT3 were measured by Real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of JAK2， phosphorylated STAT3 （p-STAT3） and STAT3 by Western blot.ResultThe highest expression of lncRNA HOTAIR was detected in HCT116 cells. Compared with the blank group， each Jianpi Xiaoai prescription group exhibited a decreased number of invasive cells （P<0.05， P<0.01）. The relative JAK2 mRNA expression in the middle-dose Jianpi Xiaoai prescription group was down-regulated （P<0.05）， and the relative lncRNA HOTAIR mRNA expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative JAK2 mRNA expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated （P<0.01）. Compared with the blank group，the relative p-STAT3 protein expression was down-regulated in the middle-dose Jianpi Xiaoai prescription group （P<0.05）， and the relative JAK2 protein expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative p-STAT3 protein expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated （P<0.01）.ConclusionJianpi Xiaoai prescription effectively inhibits the metastasis of colon cancer cells， which may be related to the inhibition of lncRNA HOTAIR/JAK2/STAT3 signaling pathway.
Abstract：ObjectiveTo investigate the mechanism of Shugan Bushen Yulin decoction in inhibiting voltage-dependent anion-selective channel protein 2 （VDAC2） gene methylation， affecting sperm mitochondrial function， and improving sperm motility through the cyclic adenosine monophosphate/protein kinase A （cAMP/PKA） pathway.MethodForty male SD rats were randomly divided into the blank group， model group， high- and low-dose Shugan Bushen Yulin decoction groups， and L-carnitine group， with eight rats in each group. Adenine （0.05 g·kg-1） was administered by gavage for 14 d for inducing oligospermia and asthenospermia. Rats in the Shugan Bushen Yulin decoction groups were treated with intragastric administration of 32.4， 8.1 g·kg-1 Shugan Bushen Yulin decoction， respectively， while those in the L-carnitine group received 0.27 g·kg-1 L-carnitine by gavage. Following the measurement of sperm motility using an automatic sperm analyzer， the pathological changes in testicular tissue were observed by hematoxylin-eosin （HE） staining. Sperm mitochondrial membrane potential was detected by flow cytometry. The expression of VDAC2 in the testicular tissue was determined by immunofluorescence assay. Real-time polymerase chain reaction （Real-time PCR） was conducted for detecting VDAC2 mRNA expression in testicular tissue. The methylation of VDAC2 gene was examined using bisulfite sequencing. The cAMP expression in testicular tissue was detected by enzyme-linked immunosorbent assay （ELISA）， and the PKA protein expression in testicular tissue by Western blot.ResultCompared with the blank group， the model group exhibited significantly decreased sperm density and motility （P<0.01）， increased mitochondrial membrane potential （P<0.01）， down-regulated VDAC2 mRNA and protein expression， PKA protein expression， and cAMP content in testicular tissue （P<0.01）， and elevated VDAC2 gene methylation （P<0.01）. Compared with the model group， L-carnitine and Shugan Bushen Yulin decoction at the high and low doses all remarkably increased the sperm density and motility and mitochondrial membrane potential （P<0.01）， up-regulated VDAC2 mRNA and protein expression， PKA protein expression， and cAMP content in the testicular tissue （P<0.01）， and lowered the methylation of VDAC2 in testicular tissue （P<0.01）. The comparison with the L-carnitine group showed that the sperm density and motility and mitochondrial membrane potential in the low-dose Shugan Bushen Yulin decoction group declined significantly （P<0.01）. The VDAC2 mRNA and protein expression， PKA protein expression， and cAMP content in the testicular tissue were significantly down-regulated （P<0.01）， while the methylation of VDAC2 was significantly enhanced （P<0.01）.ConclusionShugan Bushen Yulint decoction may inhibit VDAC2 gene methylation， increase VDAC2 expression， regulate cAMP/PKA pathway， and change mitochondrial membrane potential to enhance the sperm motility.
Abstract：ObjectiveTo investigate the synergistic effect of Xiangdan injection （XDI） and Qingkailing injection （QKLI） in the treatment of inflammation and thrombosis animal model based on changes of thrombus， inflammatory indexes， organ function， and pathological changes.MethodA total of 100 male SD rats were randomly divided into a normal control group， a model group， XDI groups （2.5， 5 mL·kg-1）， QKLI groups （5， 10 mL·kg-1）， and XDI + QKLI groups ［（2.5+5） mL·kg-1，（2.5+10） mL·kg-1，（5+5） mL·kg-1，and （5+10） mL·kg-1］ according to the body weight， with 10 rats in each group. Rats were treated correspondingly by intraperitoneal injection once a day for 4 days. The normal control group and the model group received normal saline. On the second day of administration， the model was induced in rats except those in the normal control group. Specifically， 25 mg·kg-1 carrageenan was injected intraperitoneally into the rats， followed by an injection of 50 μg·kg-1 lipopolysaccharide （LPS） through the tail vein 16 hours later. Twenty-four hours after LPS injection， the rats were detected for liver index， kidney index， the number of platelets （PLT）， thrombus length， and biochemical indicators such as aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， alkaline phosphatase （ALP）， creatinine， and blood urea nitrogen （BUN）. Enzyme-linked immunosorbent assay （ELISA） was used to determine the content of inflammatory factors interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of heart， liver， lung， and kidney， as well as the grading of organ injury.ResultCompared with the normal group， the model group showed decreased PLT， lengthened thrombus in the tail， increased liver index， elevated content of ALT， ALP， BUN， IL-6， and TNF-α （P<0.05， P<0.01）， and damaged liver， lung， and kidney tissues （P<0.05， P<0.01）. Compared with the conditions in the model group， XDI at 5 mL·kg-1 reduced serum ALT and ALP in rats （P<0.05， P<0.01）， QKLI at 5 and 10 mL·kg-1 reduced serum levels ALT and ALP， and TNF-α content （P<0.05， P<0.01）. XDI at 5 mL·kg-1 or QKLI at 10 mL·kg-1 relieved the LPS-induced lung injury （P<0.05）， the combination of XDI and QKLI decreased the levels of ALT， AST， ALP， and TNF-α， and the effect was predominant in the combination of XDI and QKLI at 5 and 10 mL·kg-1 （P<0.05， P<0.01）. Additionally， the length of the tail thrombus was significantly shortened （P<0.05）， and the degree of lung injury was also reduced （P<0.05）. The serum levels of ALT and BUN， TNF-α content， and liver index of rats were reduced after the combination of XDI and QKLI as compared with those in the single drug groups at the same dose （P<0.05， P<0.01）.ConclusionXDI or QKLI can improve or inhibit organ function， organ injury， and inflammatory response in the rat model of inflammation and thrombosis. The combination of the two drugs shows a synergistic effect in reducing the length of venous thrombus， improving liver and kidney function， inhibiting inflammatory factors， and protecting lung， liver， kidney， and other organs.
Abstract：ObjectiveTo investigate the mechanism of joint cavity injection of Dioscoreae Rhizoma polysaccharides （DRP） in protecting against cartilage degeneration and inhibiting the expression of inflammatory factors in the rabbit model of knee osteoarthritis to provide relevant references for the development and further research on DRP.MethodFifty-five New Zealand white rabbits were selected for the induction of knee osteoarthritis model by the modified Hulth's modeling method. The model rabbits were randomly divided into a model group， a sodium hyaluronate group （1.00 mg·kg-1）， and low- （0.7 mg·kg-1）， medium- （1.43 mg·kg-1）， and high-dose （2.15 mg·kg-1） DRP group according to a random number table. One week after modeling， the rabbits in the groups with drug intervention were treated correspondingly for five weeks， once per week， and no intervention was performed in the model group. Five weeks later， the joint specimens were observed by visual observation. The articular cartilage tissues were observed under the light microscope for pathological sections and scores by hematoxylin-eosin （HE） staining and toluidine blue （TB） staining. The expression levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） in the synovial fluid were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of matrix metalloproteinase-13 （MMP-13）， transforming growth factor-β1 （TGF-β1）， and type Ⅱ collagen （Col-Ⅱ） in the articular cartilage were measured by immunohistochemistry.ResultAfter five weeks of DRP intervention， compared with the model group， the DRP groups exhibited lowered levels of IL-6， IL-1β， and TNF-α in the synovial fluid （P<0.05）， reduced expression of MMP-13 in the articular cartilage （P<0.05）， and increased levels of TGF-β1 and Col-Ⅱ （P<0.05）. Compared with the low-dose and high-dose DRP groups， the medium-dose DRP group showed reduced levels of IL-6， IL-1β， and TNF-α in the knee joint （P<0.05）， increased levels of TGF-β1 and Col-Ⅱ in cartilage tissues （P<0.05）， and dwindled level of MMP-13 （P<0.05）. Compared with the sodium hyaluronate group， the medium-dose DRP group showed no significant differences in IL-6， IL-1β， and TNF-α in rabbit knee joints， and TGF-β1， Col-Ⅱ， and MMP-13 in cartilage tissues.ConclusionJoint cavity injection of DRP can significantly reduce the expression of IL-6， IL-1β， and TNF-α in rabbit synovial fluid， effectively inhibit the expression of MMP-13 in the articular cartilage to suppress the degradation of articular cartilage collagen and promote the synthesis of TGF-β1 and Col-Ⅱ. Therefore， DRP can protect and repair articular cartilage to delay the degeneration of articular cartilage.
Abstract：ObjectiveTo explore effects of different extracts and monomers of Lepidium meyenii （Maca） on the proliferation of mouse splenic lymphocytes and induction of interleukin-2 （IL-2） and tumor necrosis factor-α （TNF-α） by observing their immunomodulatory effects.MethodAn octadecylsilyl （ODS） column was used to enrich the methanol extract of L. meyenii in stages to obtain six fractions and three monomers. Different groups of extracts and monomers of L. meyenii at different doses were set up. Cell counting Kit-8 （CCK-8） was used to detect the effect on the proliferation of mitogen-free， concanavalin A （Con A）-induced， and lipopolysaccharides （LPS）-induced mouse splenic lymphocytes. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of IL-2 and TNF-α.ResultL. meyenii extracts Fr3 and Fr6， and monomers N-benzyl hexadecanamide and 1，2-dihydro-4-carboxaldehyde-3-benzyl-N-hydroxypyridine slightly promoted the proliferation of Con A-induced T lymphocytes and LPS-induced B lymphocytes （P<0.01） as compared with the conditions in the model group. L. meyenii extracts and monomers significantly induced the secretion of IL-2 and TNF-α by splenic lymphocytes （P<0.01）.ConclusionL. meyenii extracts and monomers can achieve immunological enhancement by promoting the secretion of IL-2 and TNF-α， and facilitate the proliferation of splenic lymphocytes. The active components are presumedly macamides and pyridine alkaloids， and the specific mechanism still needs to be further explored.
Abstract：ObjectiveTo investigate the effects of external application of modified Xiangsha Liujunzi Tang in improving gastrointestinal reaction，immune function， and quality of life in patients with gastrointestinal tumors after moderately/highly emetogenic chemotherapy.MethodA total of 140 inpatients （from January 2018 to May 2021） with gastrointestinal malignancies diagnosed by the Oncology Department of Integrated Traditional Chinese and Western Medicine of China-Japan Friendship Hospital and treated with moderately/highly emetogenic chemotherapy were randomly divided into an experimental group （n=70） and a control group （n=70） according to the Good Clinical Practice （GCP）. Participants were given routine antiemetic treatment once 20 minutes before chemotherapy （tropisetron 5 mg + dexamethasone 5 mg or methylprednisolone 40 mg）. On this basis， the patients in the experimental group were treated with external application of modified Xiangsha Liujunzi Tang for 1 week on the second day after chemotherapy （the selected points were Zhongwan， Neiguan and Zusanli， 6 hours a day， and the application was changed every day）， and the patients in the control group were applied with comfort patch at the same time. The gastrointestinal reaction grade， subset concentration of lymphocytes， and Karnofsky score in two groups before and one week after chemotherapy were recorded， and all data were analyzed by SPSS 19.0 statistical software.ResultThe grades of chemotherapy-related gastrointestinal reaction in two groups after treatment decreased as compared with that before treatment （P<0.05）. The experimental group was superior to the control group after treatment in terms of grade decrease （P<0.05）. The total response rate of the experimental group was 81.43% （57/70）， higher than 62.86% （44/70） in the control group （χ2=9.73， P<0.05）. Stratified analysis was performed on the experimental group. Compared with the conditions before treatment， the grade of gastrointestinal reaction in patients with esophageal cancer and colorectal cancer of the experimental group decreased after treatment （P<0.05）， and that in patients of different genders decreased after treatment （P<0.05）. The results of immune function showed that there was no significant difference in the concentrations of CD3+， CD4+， CD8+， and NK cell subsets of the control group before and after treatment， while the concentrations of CD3+， CD4+， and NK cell subsets of the experimental group were higher than those before treatment and even superior to those in the control group （P<0.05）. The Karnofsky score of quality of life in the experimental group was higher than that in the control group， with quality of life improved （P<0.05）.ConclusionExternal application of modified Xiangsha Liujunzi can improve the gastrointestinal reaction，immune function， and quality of life of patients with gastrointestinal tumors after moderately/highly emetogenic chemotherapy.
Keywords：Xiangsha Liujunzi Tang;gastrointestinal tumor;external treatment of traditional Chinese medicine;chemotherapy;gastrointestinal reaction
Abstract：ObjectiveTo evaluate the clinical efficacy and safety of Longmu Zhuanggu granule for the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency.MethodThis multicenter stratified， block-randomized， double-blind， double-dummy， positive drug （pidotimod granule） parallel controlled， and non-inferiority trail intended to included 240 children patients and divided them into the experimental group （n=120） and the control group （n=120） at the ratio of 1∶1. Patients in both groups were treated for eight successive weeks and followed up for 12 months. The cure rates， numbers of respiratory infections， average courses of disease， curative effects of traditional Chinese medicine （TCM） syndrome， curative effects of individual symptoms， curative effects of immune indexes， and safety indexes between the two groups were observed and compared.ResultA total of 237 subjects were collected from 10 research centers， including 119 cases in the control group and 118 in the experimental group. There were 236 cases enrolled into the full analysis set （FAS）， 210 into the per-protocol set （PPS）， and 236 into the safety set （SS）. The baseline data of the two groups were not significantly different from each other， indicating that they were comparable. The cure rates of the experimental group and control group were 75.21% （88/117） and 73.95%（88/119）， respectively， with the 95% confidence interval （CI） of difference between the two groups being 1.26% （-9.85%，12.37%） for FAS and 3.81% （-6.28%，13.90%） for PPS. The 95% CI fell within the 10% non-inferiority margin， implying that non-infertility test of the cure rate in the treatment of endpoint disease was valid， and the conclusions of FAS and PPS analysis were consistent. There was no significant difference in the number or course of upper respiratory infection， bronchitis， and pneumonia. The difference in curative effects of TCM syndrome between the two groups after four weeks of treatment was not remarkable. After eight weeks of treatment， the total effective rate of the experimental group was 84.62%（99/117）， statistically higher than 78.15%（93/119） of the control group（χ2=-3.26，P<0.05）. There were no significant differences in the disappearance rates of individual symptoms between the two groups after four weeks of treatment. After eight weeks of treatment， the experimental group and control group exhibited the disappearance rates of 67.50%（54/80） and 47.37%（36/76） for shortness of breath and laziness to speak， 75.00%（54/72） and 53.33%（40/75） for poor appetite， 54.55%（60/110） and 37.84%（42/111） for hyperhidrosis， respectively， with obviously better outcomes observed in the experimental group （P<0.05，P<0.01）. The inter-group comparison revealed significant differences in immune indexes after eight weeks of treatment. As demonstrated by comparison with the situations before treatment， IgA， IgG， IgM， and CD4 did not change significantly after treatment. Except for CD8 in the experimental group (P<0.05), there was no significant difference in other immune indexes before and after treatment There was no significant difference in the incidence of adverse reactions.ConclusionLongmu Zhuanggu granule is not inferior to pidomod granule in the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency， and it exhibits good safety， implying its promising clinical application value.
Abstract：ObjectiveTo investigate the effect of Qixian Tongluo prescription on neural function recovery in patients with cerebral infarction and its mechanism.MethodA total of 100 inpatients （January to June，2020）with cerebral infarction in the Neurology Department of Wenzhou Hospital of Traditional Chinese Medicine were assigned to an experimental group （n=50） and a control group （n=50） according to the random number table. Both groups received conventional treatment of western medicine，while the experimental group took additional Qixian Tongluo prescription. Treatment lasted for 12 weeks. The clinical efficacy，National Institutes of Health Stroke Scale （NIHSS） score， the modified Barthel index （MBI），Fugl-Meyer assessment （FMA） score， and levels of brain-derived neurotrophic factor（BDNF），vascular endothelial growth factor（VEGF）， and stromal cell-derived factor-1（SDF-1） in peripheral blood of the two groups before and after treatment were compared.ResultThe total response rate in the experimental group was 84.00%（42/50），higher than 66.00%（33/50） in the control group （Z=-7.365，P<0.05）. There was no significant difference in the scores of MBI，FMA， and NIHSS before treatment between the two groups. The MBI and FMA scores of the two groups increased （P<0.01）， and the NIHSS scores decreased （P<0.05， P<0.01）. Compared with the control group after treatment， the experimental group showed increased MBI and FMA scores and decreased NIHSS score （P<0.05）. There was no significant difference in BDNF level between the two groups before and after treatment. The VEGF and SDF-1 levels in the peripheral blood of the two groups were higher than those before treatment （P<0.05）， and the experimental group was higher than the control group （P<0.05）.ConclusionQixian Tongluo prescription can effectively improve the clinical efficacy，the quality of life， and the prognosis of patients with cerebral infarction during convalescence. The underlying mechanism is associated with the promotion of the expression of endogenous VEGF and SDF-1 in the peripheral blood to activate the SDF-1/chemokine receptor 4（CXCR4） signaling pathway， induce the recruitment and mobilization of endothelial progenitor cells， and facilitate the angiogenesis and repair of ischemic brain tissues.
Keywords：convalescence of cerebral infarction;Qixian Tongluo prescription;warming yang and promoting blood circulation;neural repair;brain-derived neurotrophic factor（BDNF）;vascular endothelial growth factor（VEGF）;stromal cell-derived factor-1（SDF-1）
Abstract：ObjectiveTo observe the clinical efficacy of the modified Buzhong Yiqitang combined with Erxian decoction in treating stress urinary incontinence （SUI） of perimenopausal women due to spleen and kidney Qi deficiency.MethodOne hundred and six patients were randomly divided into a control group （52 cases） and an observation group（54 cases）. Patients in both groups received lifestyle intervention and pelvic floor muscle training （PFMT）. On this basis， patients in the observation group were further treated with the modified Buzhong Yiqitang combined with Erxian decoction， 1 bag/day， while those in the control group were provided with Suoquan pills， 6 g/time， 2 times/day， for eight weeks. Following the international consultation on incontinence questionnaire-short form （ICIQ-SF） scoring before and after treatment， the urodynamic parameters such as maximum urinary flow rate （Qmax）， maximum urethral closure pressure （MUCP）， residual urine volume （RUV）， abdominal pressure leakage point pressure （ALPP）， and bladder capacity （BC） were measured. The number of incontinence episodes per 24 h， the degree of urinary incontinence， the amount of 1 h urine leakage， and the spleen and kidney Qi deficiency syndrome score were recorded before and after treatment. The levels of estradiol （E2）， follicle stimulating hormone （FSH）， pituitary adenylate cyclase activating peptide （PACAP）， and vasoactive intestinal peptide （VIP） were measured before and after treatment.ResultThe ICIQ-SF sub-scores of the urinary incontinence frequency， severity， and impact on quality of life as well as the total score in the observation group were all lower than those in the control group （P<0.01）. Qmax， MUCP， ALPP and BC in the observation group were elevated in contrast to those in control group （P<0.01）， while the RUV declined （P<0.01）. Compared with the control group， the observation group exhibited a decreased number of incontinence episodes per 24 h， milder degree of urinary incontinence， reduced amount of 1 h urine leakage， and lower spleen and kidney Qi deficiency syndrome score （P<0.01）. The E2， PACAP， and VIP in the observation group were up-regulated as compared with those in the control group （P<0.01）， whereas the FSH was down-regulated （P<0.01）. The cure and effective rates of the observation group were （29/50） 58.00% and （47/50）94.00%， respectively， significantly better than （18/48）37.50% and （38/48）79.17% of the control group （χ2=4.124， χ2=4.683， P<0.05）.ConclusionOn the basis of the lifestyle intervention and PFMT， the modified Buzhong Yiqitang combined with Erxian decoction obviously alleviates urinary incontinence， adjusts sex hormones， PACAP and VIP， ameliorates urodynamic parameters， and enhances the quality of life of patients with SUI due to spleen and kidney Qi deficiency. The resulting cure and effective rates are superior to those of the positive control.
Abstract：ObjectiveTo observe the clinical efficacy and safety of modified Xiao Chengqitang combined with acupoint catgut implantation in treating diet-induced obesity （DIO） syndrome of stomach heat dampness obstruction.MethodOne hundred and seventy-two patients were randomly divided into control group（84 cases） and observation group（88 cases）. Both groups of patients received diet and exercise lifestyle adjustments， and acupoint catgut implantation was performed， 10 days for 1 time， 5 days intervals and then catgut implantation again， for a total of 6 times. Patients in observation group took modified Xiao Chengqitang granular powder， 10 g/time， with lukewarm boiled water in morning and evening. Patients in control group took modified Xiao Chengqitang granular powder simulant， 10 g/time， with lukewarm boiled water， 2 times/day. The treatment courses continued 4 months in two groups. Then the body mass index （BMI）， fat percentage （F%）， obesity， waist to hip ratio （WHR） were measured before and after treatment. Color Doppler ultrasonography was used to measure abdominal fat thickness， prehepatic fat thickness （AHF）， perirenal fat thickness （PRF）， and visceral fat index （UVI）. Fasting blood glucose （FBG）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein-cholesterol （HDL-C）， low-density lipoprotein-cholesterol （LDL-C）， fasting insulin （FINS）， leptin （LP）， and adiponectin （APN） were detected before and after treatment， and the homeostasis model assessment of insulin resistance （HOMA-IR） index was calculated. In addition， safety evaluation was also conducted.ResultThe BMI， F%， obesity degree and WHR in observation group were all lower than those in control group （P<0.05 or P<0.01）. Subcutaneous fat thickness， AHF， PRF and UVI in observation group were lower than those in control group （P<0.01）. The TG， TC， LDL-C and FINS levels in observation group were lower than those in control group （P<0.01）. The LP and HOMA-IR were also lower than those in control group （P<0.01）， while the APN was higher than that in control group （P<0.01）. The total effective rate in clinical application was （71/80） 88.75% in the observation group， higher than （57/75） 76.00% in the control group （χ2=4.374， P<0.05）.ConclusionModified Xiao Chengqitang combined with acupoint catgut implantation in treating DIO syndrome of stomach heat dampness obstruction can adjust LP， APN and other factors， improve energy metabolism such as sugar and fat， and effectively control obesity with high safety， so it is worthy of clinical use.
Abstract：ObjectiveTo compare the effects of different drying methods on the chemical constituents of Trichosanthis Fructus.MethodTrichosanthis Fructus was dried by means of air drying， sun drying， hot air drying （40， 60， 80 ℃） and variable temperature drying （50-80， 80-50 ℃）. The contents of nucleosides and flavonoids in Trichosanthis Fructus peels and seeds treated by different methods were compared by high performance liquid chromatography （HPLC）， mobile phase was acetonitrile-0.2% acetic acid aqueous solution （3∶7） （A）-acetonitrile （B） for gradient elution （0-15 min， 97-95%B； 15-30 min， 95%-90%B； 30-35 min， 90%-87%B； 35-40 min， 87%-86.5%B； 40-48 min， 86.5%-97%B； 48-50 min， 97%B）， the detection wavelength was 260 nm， and the flow rate was 0.4 mL·min-1. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to compare the changes of volatile components in the samples treated by different treatments. The volatile components were incubated on a SE-54 capillary column （0.32 mm×30 m， 0.25 μm） at 80 ℃ and 500 r·min-1 for 15 min， the injection temperature was 85 ℃， the injection volume was 400 μL， the analysis time was 35 min， carrier gas was high purity nitrogen， the flow rate of carrier gas was 2.0 mL·min-1， the flow rate of drift gas was 150 mL·min-1， and the temperature of IMS detector was 45 ℃.ResultThe contents of uridine， adenosine and adenine were higher after hot air drying at >50 ℃. Low temperature drying was conducive to maintaining the stability of cytidine， cytosine， rutin， luteolin and 2ʹ-deoxyadenosine. GC-IMS technology could realize the analysis and identification of Trichosanthis Fructus samples after different treatments. There were more volatile components after hot air drying at 80 ℃ and variable temperature drying.ConclusionHot air drying at 40 ℃ and 60 ℃ can retain nucleosides and flavonoids， and the volatile components are similar to those in traditional drying methods， which has the advantages of high efficient， controllable and suitable for industrial production.
Keywords：hot air drying;Trichosanthis Fructus;nucleosides;volatile components;gas chromatography-ion mobility spectrometry （GC-IMS）;flavonoids
Abstract：ObjectiveThe biological mechanism of Codonopsis pilosula adaptation to drought was explored by determining the root metabolome of C. pilosula during harvesting.MethodNon-targeted metabonomics LC-MS was used to screen differential metabolites by multivariate statistical analysis，univariate statistical analysis and metabolic pathway enrichment analysis.Result①There were 274 metabolites in LD vs CK group，142 of which were up-regulated and 132 of which were down-regulated. There were 284 metabolites with significant difference in MD vs CK group，of which 157 were up-regulated and 127 were down-regulated. There were 317 metabolites with significant difference in SD vs CK group，of which 133 were up-regulated and 184 were down-regulated. ②Differential metabolites were annotated into kyoto Encyclopedia of Gene and Genomes （KEGG） database and 82 differential metabolic pathways were obtained，among which sphingolipids metabolism was significantly enriched （P<0.01）.Metabolism of arginine and proline，tryptophan，alanine，galactose，nicotinic acid and nicotinamide，cysteine and methionine，arachidonic acid，linolenic acid and glycerides were significantly enriched in different metabolite pathways （P<0.05）. ③The metabolites of the three comparison groups before and after enrichment were classified and analyzed. It was found that they were mainly concentrated in fatty acyls group，carboxylic acid and derivatives，and organ oxygen compounds，followed by sphingolipids，indoles and derivatives，organonitrogen compounds，glycerophospholipids，pyridines and derivatives，peptidomimetics，glycerolipids and so on.In the drought stress of C. codonopsis，carbohydrate related metabolites were mainly up-regulated，lipid related metabolites were mainly down-regulated，and all other metabolites were up-regulated.ConclusionThe changes of metabolites in the roots of C. pilosula under drought stress were elucidated. carbohydrate and lipid-related metabolites were the main products of C. pilosula under drought stress，and these metabolites may be the main reason to improve the ability of C. pilosula to resist drought，which laid a foundation for further study on the mechanism of C. pilosula to resist drought.
Abstract：ObjectiveTo develop the specific molecular markers of Codonopsis plants and better identify their germplasm resources considering the significant difference in active ingredients of Codonopsis Radix from various origins and producing areas.MethodSuch bioinformatics software as Primer 5.0， NTSYS-pc 2.10e， and PopGene 32 were used for searching the simple sequence repeat （SSR） markers of C. minima chloroplast genome， C. tsinlingensis chloroplast， and C. lanceolata mitochondrial sequences， and 120 pairs of SSR primers were designed by Primer 5.0. Then 16 pairs of cpSSR primers and 10 pairs of mtSSR primers with good screening effect and high polymorphism were selected for analyzing the interspecific versatility of 20 samples.ResultThe results showed that 66 cpSSR primer sites and 26 mtSSR sites were identified from the genome sequences， with 86.20% of single nucleotide， 6.9% of dinucleotide， and 6.9% of trinucleotide for C. minima chloroplast， 83.78% of single nucleotide，13.51% of dinucleotide， and 2.71% of trinucleotide for C. tsinlingensis chloroplast， and 46.15% of single nucleotide and 53.85% of dinucleotide for C. lanceolata mitochondria. As demonstrated by polymerase chain reaction （PCR） identification results， the developed 26 pairs of SSR primers had good applicability in the genus Codonopsis. The analysis by NTSYS-pc 2.10e revealed that the genetic similarity coefficients of 20 samples were within the range of 0.38-1.00， and they were divided into two subgroups at a threshold of 0.69. Four pairs of polymorphic primers were screened out in the diversity analysis of 20 samples using PopGene 32. The number of observed alleles （Na） was 12， and the effective number of alleles （Ne） ranged from 1.362 9 to 2.605 9. The percentage of polymorphic loci （PPL） at each site was 100%， and the average values of genetic parameters Ho， He， and I at each site were 0.555 8， 0.444 2， and 0.753 2， respectively， indicating high polymorphism at each site. The screened four pairs of primers were utilized for DNA fingerprinting of the 20 samples， and it was found that the DNA fingerprints enabled the identification of these 20 samples.ConclusionThis study has provided a molecular basis for the study of the genetic relationship between plants in species Codonopsis and the intraspecific genetic differentiation.
Keywords：Codonopsis;chloroplast;mitochondria;microsatellite;development and application
Abstract：ObjectiveTo establish the sequence-related amplified polymorphism （SRAP）-polymerase chain reaction （PCR） system for Valeriana officinalis var. latifolia，so as to lay the theoretical and technical foundations for the breeding of V. officinalis var. latifolia.MethodSingle factor test was applied to investigate the effects of Taq Mix dose，Mg2+ concentration，template DNA concentration，and Taq DNA polymerase content on SRAP-PCR amplification of V. officinalis var. latifolia，based on which the orthogonal experiments were performed to optimize the SRAP-PCR system for V. officinalis var. latifolia. The effective primers that could be used for genetic diversity studies of V. officinalis var. latifolia were selected under the optimal reaction condition.ResultThe results of the single factor test showed that Taq Mix dose within the range of 8-11 μL resulted in better amplification. The addition of a low concentration of Mg2+，the medium to low concentrations of template DNA，or the low concentration of Taq DNA polymerase enhanced the amplification efficiency or richness. As demonstrated by the orthogonal experiments，the influencing degrees of related factors on SRAP-PCR amplification of V. officinalis var. latifolia were sorted in a descending order as follows： Taq Mix dose>Taq DNA polymerase content>Mg2+ concentration>template DNA concentration. The optimal reaction system for V. officinalis var. latifolia was determined to consist of 11 μL of Taq Mix，30 ng of template DNA，0.025 mmol·L-1 Mg2+，1.5 U of Taq DNA polymerase，5 μmol·L-1 forward primer，and 5 μmol·L-1 reverse primer，which was supplemented to 20 μL with ddH2O. The optimal annealing temperature was 36.8 ℃. A total of 17 pairs of effective primers with high band resolution and polymorphism were selected from 88 primer pairs for SRAP-PCR of V. officinalis var. latifolia.ConclusionThe established SRAP-PCR system for V. officinalis var. latifolia is stable， which can be used for genetic diversity studies of V. officinalis var. latifolia.
Keywords：Valeriana officinalis var. latifolia;sequence-related amplified polymorphism （SRAP）;system optimization;primer screening;orthogonal design
Abstract：ObjectiveTo explore the potential suitable distribution area and the high-quality distribution area of Sabia parviflora.MethodCombined with the distribution information and environmental factors，the maximum entropy （MaxEnt） model and ArcGIS software were used to predict the potential suitable distribution area of S. parviflora. Based on the correlation between environmental factors and total saponins，total flavonoids，quercetin-3-O-gentiobioside，camellianoside，tsubakioside A，kaempferol-3-O-rutinoside and isobariclisin-3-O-rutinoside，the quality regionalization was conducted by using spatial interpolation method and fuzzy superposition function in ArcGIS software.ResultS. parviflora is mainly distributed in Yunnan，Guizhou，Guangxi province in China. The medium and high suitable areas accounts for about 2.88% of the national area. The precipitation in October and November，the precipitation in the warmest and driest seasons，the standard deviation of seasonal changes in temperature and altitude are the main environmental factors that affect the distribution of S. parviflora. Slope，precipitation，solar radiation and temperature change had great influence on the accumulation of secondary metabolites. Based on the results of potential suitable distribution and spatial interpolation of each component，the high-quality areas of S. parviflora are mainly concentrated in the southwest of Guizhou，with Qinglong，Guanling，Zhenning，Pu'an，Xingren county and other areas as the core.ConclusionThis study provides a scientific guidance for the site selection of artificial planting and the procurement of medicinal materials for S. parviflora.
Abstract：ObjectiveTo select and evaluate new Chinese herbal prescription for the treatment of decreased ovarian reserve （DOR） and its appropriate dosage.MethodThe literature concerning the treatment of DOR with traditional Chinese medicine （TCM） was retrieved from such databases as Chinese Journal Full-text Database， Wanfang Data Knowledge Service Platform， and Chongqing Weipu Database for Chinese Technical Periodicals （VIP）， based on which a database was established using the Traditional Chinese Medicine Inheritance Support System （TCMISS） V2.5. The data mining was then carried out to obtain the core combinations of Chinese herbs and new Chinese herbal prescription combinations， followed by the determination of the new Chinese herbal prescription by expert group discussion for experiment evaluation. The female SD rats were divided into the normal group， DOR model group， Kuntai capsule group， and low-， medium-， and high-dose new Chinese herbal prescription groups， with 12 rats in each group. Rats in the Kuntai capsule group and low-， medium-， and high-dose new Chinese herbal prescription groups were treated with Kuntai capsule solution （0.5 g·kg-1 determined according to the dosage in the instruction） and 3.037 5， 6.075， and 9.12 g·kg-1 new Chinese herbal prescription， respectively. After 21 days， the estrous cycle was observed by vaginal exfoliated cell smear， and the ovarian structure was observed by hematoxylin-eosin （HE） staining. The serum anti-mullerian hormone （AMH）， follicle stimulating hormone （FSH）， luteinizing hormone （LH）， and estrogen （E2） contents as well as the reactive oxygen species （ROS）， malondialdehyde （MDA）， and glutathione （GSH） levels and superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） activities in ovary were detected using biochemical methods.ResultThe new Chinese herbal prescription subjected to experimental evaluation was composed of 11 Chinese herbs， namely Rehmanniae Radix Praeparata 20 g， Cervi Cornus Colla 12 g， Lycii Fructus 20 g， Corni Fructus 12 g， Albiziae Cortex 9 g， Nelumbinis Plumula 3 g， Salviae Miltiorrhizae Radix et Rhizoma 20 g， Astragali Radix 30 g， Atractylodis Macrocephalae Rhizoma 12 g， Dioscoreae Rhizoma 30 g， and Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle 6 g. Compared with the model group， the Kuntai capsule group and medium- and high-dose new Chinese herbal prescription groups exhibited significantly improved estrous cycle and follicular development， elevated serum AMH and E2 and ovarian GSH （P<0.05）， decreased serum FSH and LH （P<0.05） and ovarian ROS and MDA （P<0.05）， and enhanced SOD， CAT， and GSH-Px activities （P<0.05）. There were no significant differences in the above-mentioned indexes between the Kuntai capsule group and the middle- and high-dose new Chinese herbal prescription groups， but the estrous cycle and follicular development were better in the latter two groups.ConclusionThe new Chinese herbal prescription screened by data mining is able to enhance ovarian antioxidation， promote follicular development， ameliorate serum hormone and estrous cycle， and effectively improve ovarian reserve function in DOR rats. The medium dose （6.075 g·kg-1） has been proved optimal.
Keywords：decreased ovarian reserve （DOR）;rats;hormones;antioxidation;Traditional Chinese Medicine Inheritance Support System （TCMISS）
Abstract：Airway mucus is an important part of the defense barrier function of the airway. Abnormal secretion of airway mucus is closely related to recurrent attacks， delay or aggravation of respiratory infectious or infectious diseases such as chronic obstructive pulmonary disease （COPD）， community-acquired pneumonia， and coronavirus disease 2019 （COVID-19）. Lianhua Qingke tablets， an innovative traditional Chinese medicine （TCM） developed under the guidance of the theory of collateral disease of TCM， has the function of reducing phlegm and relieving cough， which can reduce the generation and viscosity of sputum and promote sputum excretion. Clinical studies have shown that it can significantly improve the symptoms of expectoration and cough in patients with acute bronchitis or COVID-19， confirming its scientific connotation and clinical value of reducing phlegm and relieving cough to improve ventilation and exchanging function.
Keywords：airway mucus;collateral-disease theory;Lianhua Qingke;reducing phlegm;disease of respiratory system;coronavirus disease 2019（COVID-19）
Abstract：Daturae Flos is a traditional antitussive and antiasthmatic medicine， its flowers and leaves are rich in a variety of compounds， including withanolides， alkaloids， terpenes， flavonoids and amides. Because of its antiasthmatic， antitussive， antispasmodic and analgesia effect， it is traditionally used for the treatment of asthma， cough， cold pain in abdominal cavity， rheumatic arthralgia， infantile chronic eclampsia， and can also be used as raw material for surgical anesthesia. Modern pharmacological studies have shown that in addition to the traditional efficacy， Daturae Flos also has anti-inflammatory， immunosuppression， anti-convulsion and other effects， and is often used in the treatment of psoriasis， Parkinson's disease， epilepsy， rheumatoid arthritis and other diseases. At present， the chemical constituents of Daturae Flos are mainly focused on withanolides and alkaloids. At the same time， there is a lack of clear classification of chemical components and the distribution of chemical components in medicinal parts of this medicine， and little information is available for the pharmacological effects of polysaccharides. Based on this， this paper systematically searched relevant literature of Daturae Flos， and summarized and analyzed its chemical composition， pharmacological effect and clinical application， in order to provide reference for further development and utilization of Daturae Flos.
Keywords：Daturae Flos;withanolides;terpenoids;anesthesia;anti-asthmatic and antitussive;psoriasis;anti-inflammatory
Abstract：Calycosin （CA）， a functional phytoestrogenic isoflavone extracted from Chinese herb Astragali Radix， is characterized by high efficiency， low toxicity， and multiple targets and has multiple pharmacological activities such as anti-oxidation， anti-radiation， anti-bacteria， cardio-cerebrovascular protection， and immunity enhancement. A number of studies have proved its significant anti-tumor effect， making it expected to become a potential component for the treatment of malignant tumors. Research shows that CA exerts the anti-tumor effect via multiple mechanisms like inducing tumor cell apoptosis and inhibiting tumor cell proliferation， migration， and invasion. It has been proved to be effective in suppressing breast cancer， colorectal cancer， lung cancer， cervical cancer， ovarian cancer， nasopharyngeal cancer， and other common malignant tumors. Its anti-tumor activity is mainly related to the regulation of B-cell lymphoma-2 （Bcl-2） family genes， microRNA （miRNA）， and estrogen receptor β （ERβ） to trigger tumor cell apoptosis. Its anti-proliferation activity is mainly reflected in the regulation of cyclin family， WD repeat-containing protein 7 （WDR7-7）， and Ewing sarcoma-associated transcript 1 （EWSAT1）. By blocking the epithelial mesenchymal transformation （EMT）， vascular endothelial growth factor （VEGF）， matrix metalloproteinases （MMPs）， CA inhibits tumor cell metastasis and invasion. In addition， it inhibits tumors by regulating autophagy marker Beclin-1 induced tumor cell autophagy and increases the sensitivity to chemotherapy drugs， thus improving the treatment effect. Although there are many reports about the wide range of applications and good effects of CA in anti-tumor， the systematic review of its anti-tumor mechanism is still lacking. Therefore， this study reviewed the anti-tumor effects and mechanisms of CA， aiming to provide reference for researchers and clinical workers.
Abstract：Rotavirus enteritis （RVE）， a common infectious disease of the digestive system， is commonly found in infants and young children. Its incidence is high， posing great threats to human health. At present， no specific drugs are available in western medicine and the symptomatic treatments like fluid infusion， anti-diarrhea， correction of electrolyte disorder， protection of gastrointestinal mucosa， and regulation of intestinal microecology have been employed. In addition to the poor therapeutic efficacy， they do not perform well in preventing the onset of RVE and shortening the course of disease. Traditional Chinese medicine （TCM） exerts the therapeutic effect against RVE via multiple targets， without inducing obvious adverse reactions and the cost is low. The specific mechanism of action has yet to be fully explained. Although there are some studies exploring the role of Chinese medicinal monomers in the pathogenesis of RVE， the types involved are still not enough. The effects of Chinese medicinal monomers on autophagy-related pathways fail to be uncovered， which is attributed to the lack of large-scale experimental data and clinical evidence. There are also many problems that cannot be ignored in the related research of Chinese medicinal compounds， taking the superficial exploration and limited scope for instance. Autophagy is a highly conserved biological phenomenon that involves a variety of signaling pathways. Its dysfunction is related to multiple pathological processes. Studies have shown that autophagy plays an important role in the pathogenesis of RVE， especially in the early stage of viral infection. Autophagy induces intestinal mucosal barrier damage， intestinal nerve dysfunction， and immune abnormality， resulting in the occurrence and development of RVE. In recent years， a large number of experimental studies have confirmed that TCM fights against circulatory， respiratory， digestive， and immune system diseases as well as tumors by intervening in autophagy. At the same time， a handful of studies have suggested that Chinese medicinal monomers and compounds regulate autophagy and interfere with viral replication by affecting related signaling pathways， thus playing a positive role in reversing the progression of RVE. However， at present， there are few studies on the regulation of autophagy by TCM in the treatment of RVE， and no systematic elaboration is available. This review aimed to summarize the role of autophagy in the pathogenesis of RVE and its intervention with TCM， in order to provide more theoretical and clinical evidence for the treatment of RVE with TCM.
Keywords：rotavirus enteritis;autophagy;Chinese medicine compound;Chinese medicine monomer;research progress
Abstract：Lung cancer， a malignancy with high incidence rate and mortality rate， is a major threat to human life and health. At present， the common methods for the treatment of lung cancer include surgical resection， radiotherapy， chemotherapy， targeted therapy， and immunotherapy， but these methods generally have the problems of severe toxic/side effect and high treatment cost. Traditional Chinese medicine（TCM） has a history of more than 2 000 years of application in China and has its unique advantages in the treatment of tumors. Modern pharmacological experiments have found that TCM can inhibit tumor growth， prolong patients' survival， and improve clinical symptoms and patients' quality of life by inducing tumor cell apoptosis， inhibiting tumor angiogenesis， and reducing tumor cell drug resistance. Apoptosis is a process of spontaneous programmed cell death， which is closely related to the occurrence and development of the tumor. Studies have shown that many Chinese medicines can inhibit the development of lung cancer by inducing apoptosis. This study searched， analyzed， and summarized the available papers on the mechanism of TCM in the treatment of lung cancer by inducing apoptosis. It is found that Chinese medicine induces lung cancer cell apoptosis mainly by regulating apoptosis-related factors and apoptosis-related signaling pathways ［inhibitor of apoptosis proteins （IAPs）， B cell lymphoma-2 （Bcl-2）， p53 protein， the second mitochondria-derived activator of caspase （SMAC）/direct IAP-binding protein with low isoelectric point （DIABLO）， extrinsic apoptotic pathway， endogenous mitochondrial pathway， Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. In addition， the Wnt/β-catenin/survivin signaling pathway and the Notch signaling pathway also play an important role in inducing apoptosis.
Abstract：Chronic obstructive pulmonary disease （COPD）， a common clinical chronic respiratory disease， has a long course and is intractable. It is closely related to many factors， such as immune imbalance. Helper T cell 17 （Th17）， an immune-promoting cell， and regulatory T cell （Treg）， an immunosuppressive cell， maintain the balance of the immune microenvironment together. In the course of COPD， the proportion of Th17 cells usually increases， while the proportion of Tregs that inhibit Th17 activity decreases. Their coordination and balance are critical in the inflammatory and immune processes of COPD. At present， COPD is mainly treated with nasal inhalation preparations and oral drugs by western medicine. In spite of a certain therapeutic effect， side effects of drugs and heavy economic burden are becoming increasingly prominent. Modern research shows that traditional Chinese medicine （TCM） has the characteristics of few side effects， stable curative effect， and multi-target regulation， and it is advantageous and promising in the prevention and treatment of COPD. In recent years， a large number of TCM clinical and experimental trials on the intervention of Th17/Treg balance in COPD have been launched. Substantial pieces of evidence confirm that the intervention of Th17/Treg balance is an important potential target of TCM in the treatment of COPD. This study reviewed the previous research on the intervention effect of single Chinese medicine， effective components of Chinese medicine， and Chinese medicinal compound on Th17/Treg balance in COPD to comprehensively reveal the potential target of Th17/Treg balance in COPD for clinicians and scientific researchers， promote relevant research， and provide references for the rational application of TCM in the prevention and treatment of COPD.
Keywords：traditional Chinese medicine;helper T cell 17 （Th17）/regulatory T cell （Treg）;chronic obstructive pulmonary disease;research progress
Abstract：Nonalcoholic fatty liver disease （NAFLD） is a metabolic stress-induced liver injury characterized by excessive lipid accumulation in hepatocytes， which is closely related to insulin resistance and genetic susceptibility. It falls into the category of "liver lump" in traditional Chinese medicine （TCM）. NAFLD affects about 25% of the population worldwide and has become a major burden of the world health care system. However， its exact pathogenesis remains unclear. Conducting the basic research on NAFLD is of great clinical significance and social value. As an important tool for NAFLD research， animal model plays a particularly important role in clarifying the pathophysiological mechanism of NAFLD. In recent years， the modeling methods for NAFLD in China and abroad have been constantly updated， and in particular， certain progress has been made in the duplication of TCM syndrome models. By consulting and sorting out the relevant literature published in recent years in China and abroad， the author summarized the replication methods of NAFLD animal models. This paper reviewed the advantages and disadvantages of models established via dietary induction （high-fat feed， high-fat and high-fructose feed， high-fat and high-cholesterol feed， and methionine choline-deficient feed）， models with genetic defects ［leptin-deficiency （Lepob/Lepob）， autosomal recessive diabetes gene homozygous deficiency （ob/ob）， Alms1 gene （foz/foz） mutation， and FATZO mice］ and exposure to special diets， and models for TCM syndromes （liver depression and spleen deficiency syndrome， phlegm-dampness syndrome， blood stasis syndrome， combined phlegm and stasis syndrome， and qi stagnation and blood stasis syndrome）， in order to provide reference for the preparation of more scientific， reasonable， economical， and convenient animal models of NAFLD， thus laying a foundation for in-depth study of the pathogenesis， prevention， and treatment of NAFLD.
Keywords：nonalcoholic fatty liver disease （NAFLD）;animal models;dietary induction;genetic defects;traditional Chinese medicine （TCM） syndromes;research progress