Study on Mechanism of Sanhuang Decoction in Alleviating Ulcerative Colitis in Mice under Candida albicans Colonization Based on Dectin-1-Syk-CARD9 Signaling Pathway / 中国中医药信息杂志

Objective To study the mechanism of Sanhuang Decotion in the treatment of ulcerative colitis(UC)under Candida albicans colonization in mice based on Dectin-1-Syk-CARD9 signaling pathway.Methods Mice model of UC with fungal colonization were established with dextran sodium sulfate free drinking and C.albicans intragastric administration.Mice were divided into normal control group,model group,sulfasalazine group,fluconazole group,and Sanhuang Decotion low-and high-dosage groups,and receive corresponding drug interventions.General state of mice were observed,and the disease activity index(DAI)score of mice were calculated.The load of C.albicans in intestine was detected,the length of the colon was measured,and pathological scoring of the colon tissue was performed.The ultrastructural changes of colon epithelium were observed under transmission electron microscopy.The contents of TNF-α,IL-6 and IL-12 in serum and colon tissues were detected by ELISA.The mRNA and protein expression of Dectin-1,Syk,CARD9,NF-κBp65 and inflammation factors in intestinal epithelial cells and colon tissues were detected by qPCR,Western blot and immunohistochemistry.Results Compared with the normal control group,the model group mice showed reduced activity,decreased food intake,accompanied by loose stools,significantly increased DAI score,increased load of C.albicans in the intestine,shortened colon length,and increased histopathological score,with widening of gap between colon epithelial cells,cytoplasmic dissolution,mitochondrial swelling;TNF-α,IL-6 and IL-12 in serum and colon tissue increased,the expressions of Dectin-1 and CARD9 mRNA and protein in colon epithelial cells increased,p-Syk,p-NF-κBp65,CARD9,TNF-α,IL-1β,IL-6 protein expression in colon tissue increased(P<0.01,P<0.05).Compared with the model group,the Sanhuang Decotion high-dosage group mice showed a significant decrease in DAI score,decreased intestinal C.albicans load,increased colon length,decreased histopathological score,more complete and orderly arrangement of microvilli in colon epithelial cells,mild mitochondrial swelling,TNF-α,IL-6 and IL-12 in serum and colon tissue decreased,and the mRNA and protein expression of Dectin-1 and CARD9 in colon tissue increased,the expression of p-Syk,p-NF-κBp65,CARD9,TNF-α,IL-1β,IL-6 protein in colon tissue decreased(P<0.01,P<0.05).Conclusion Sanhuang Decotion may exert an anti C.albicans colonization UC effect by inhibiting the Dectin-1-Syk-CARD9 signaling pathway and reducing the release of inflammatory factors.

Mechanism of Sanhuang Xiexintang in Protecting Stress Gastric Ulcer in Rats / 中国实验方剂学杂志

ObjectiveTo explore the molecular mechanism of Sanhuang Xiexintang （SHXXT） in protecting stress gastric ulcer （SGU） in rats through network pharmacology， molecular docking， and animal experiments. MethodThe active ingredients and corresponding targets in SHXXT were collected and screened from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Traditional Chinese Medicine Information Database （TCMID）， Bioinformation Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）， and Swiss Target Prediction database. SGU-related targets were screened from the Online Mendelian Inheritance in Man （OMIM）， Therapeutic Target Database （TTD）， GeneCards database， and PharmGKB database. Herbal-ingredient-target （H-C-T） network was constructed by using Cytoscape 3.9.1 software. Protein-protein interaction （PPI） of drug and disease intersection targets was analyzed by using the Protein Interaction Platform （STRING） database. Gene ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis were conducted through the Database for Annotation Visualization and Integrated Discovery （DAVID）. The active ingredients and key targets were validated using AutodockVina 1.2.2 molecular docking software， and the experimental results were further validated through animal experiments. ResultThe 55 active ingredients were screened， and 255 potential target genes for SHXXT treatment of SGU were predicted. The PPI analysis showed that protein kinase B （Akt）， phosphatase and tensin homolog deleted on chromosome ten （PTEN）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and cyclooxygenase-2 （COX-2） are the core targets of SHXXT for protecting SGU. GO and KEGG analyses showed that SHXXT may affect the development of SGU by regulating various biological processes such as the phosphoinositide 3-kinase （PI3K）/Akt signaling pathway and inflammatory processes. The molecular docking results showed that both the active ingredients and key targets had good binding ability. Animal experiments showed that compared with the blank group， the ulcer index （UI） of the model group was significantly increased （P<0.01）， and the serum levels of TNF-α and IL-1β significantly increased （P<0.01）. The phosphorylation level of PTEN in gastric mucosal tissue was significantly down-regulated （P<0.05）. The phosphorylation levels of PI3K， Akt， and nuclear factor kappa-B （NF-κB） were significantly up-regulated （P<0.05）. Compared with the model group， the UI of the treatment group was significantly reduced （P<0.01）， and the serum levels of TNF-α and IL-1β were significantly reduced （P<0.01）. The phosphorylation level of PTEN in gastric mucosal tissue was significantly up-regulated （P<0.01）， and the phosphorylation levels of PI3K， Akt， and NF-κB were significantly downregulated （P<0.01）. ConclusionThe application of network pharmacology prediction， molecular docking simulation， and animal experimental validation confirms that SHXXT regulates the PI3K/Akt/NF-κB signaling pathway to regulate the inflammatory response of rats and thus protects the gastric mucosa of SGU rats.

Effects of Modified Sanhuang Powder （加味三黄散） Combined with Cold Compression on Swelling and Pain after Knee Arthroscopy： A Retrospective Cohort Study / 中医杂志

ObjectiveTo compare the effects of topical application of Modified Sanhuang Powder （加味三黄散， MSP） combined with cold compression versus cold compression alone on swelling and pain after knee arthroscopy through a retrospective cohort study. MethodsMedical records of 134 patients with knee arthroscopy-induced knee swelling and pain were divided into non-exposure group （51 cases） and exposure group （83 cases） based on whether they used MSP for external application after surgery. The non-exposure group received simple cold compression therapy in addition to functional exercise and routine treatment after surgery， while the exposure group received topical MSP on the basis of what were given in the non-exposure group. The Visual Analog Scale （VAS） scores were compared between the two groups before and 7 days after treatment， and knee swelling measurements were taken before and 3， 5， and 7 days after treatment. The clinical effective rate was compared between the two groups. ResultsThe VAS scores in both groups were lower after treatment （P＜0.05）， and the exposure group had lower scores than the non-exposure group （P＜0.05）. On the 3rd， 5th， and 7th days of treatment， the scores on swelling at 2 cm above the superior pole of the patella， at the midline of the patella， and 5 cm below the inferior pole of the patella significantly decreased after treatment in both groups （P＜0.05）， and the exposure group had lower scores than the non-exposure group （P＜0.05）. The total clinical effective rate in the exposure group was 91.56% （76/83）， which was higher than 78.43% （40/51） in the non-exposure group （P＜0.05）. ConclusionTopical application of MSP combined with cold compression is effective in relieving postoperative swelling and pain after knee arthroscopy and is superior to cold compress alone.

Effects of Yinqi Sanhuang Jiedu Decoction （茵芪三黄解毒汤） on Liver Hardness and Epithelial Mesenchymal Transition in Hepatic Fibrosis Model Mice / 中医杂志

ObjectiveTo investigate the possible mechanism of compound Yinqi Sanhuang Jiedu Decoction （茵芪三黄解毒汤， YSJD） against the progression of hepatic fibrosis （HF）. MethodsThirty-eight C57BL/6J mice were randomly divided into blank group， model group， silybin group and low-， medium- and high-dose YSJD groups， with eight mice in the model group and six mice each in other groups. Except for the blank group， all mice were injected intraperitoneally with 10% carbon tetrachloride （CCl4） to establish a HF model， twice a week for 8 weeks. The drug intervention started one week after the first modeling； the low-， medium- and high-dose YSJD groups were given 8.325， 16.65 and 33.3 g/（kg·d） of YSJD suspension by gavage， respectively， while the silybin group was given 55 mg/（kg·d） of silybin suspension by gavage， and the blank group and the model group were given 0.2 ml normal saline by gavage， all for 8 weeks. The liver hardness of living mice was observed using a small animal ultrasound detector， and grey-scale ultrasound was recorded. The liver tissue was observed by Sirius scarlet staining， and the proportion of collagen fiber deposition was calculated. Liver function indicators including alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， and serum laminin （LN）， hyaluronic acid （HA）， pre-collagen type III （PCIII） and collagen type IV （CIV） were also detected. The protein expression of transforming growth factor β1 （TGF-β1）， Vimentin， E-cadherin， α smooth muscle actin （α-SMA） in liver tissue were detected. ResultsCompared to the blank group， the model group showed increased gray value， collagen deposition，serum ALT， AST， HA， LN and PCIII levels， decreased expression of E-cadherin， and increased expression of N-cadherin，α-SMA，Vimentin and TGF-β1 in liver tissues （P＜0.05）. Compared to the model group， the ultrasonic gray value and the proportion of collagen fiber deposition in liver of silybin group and YSJD medium- and high-dose groups decreased， and the serum ALT， AST， LN， HA and PCⅢ levels decreased. Compared to the model group， the expression of E-cadherin in liver tissues of silybin group and all three YSJD groups increased， while the expressions of N-cadherin， Vimentin， TGF-β1 and α-SMA decreased （P＜0.05）， and among them， most improvements were seen in the medium-dose YSJD group （P＜0.05）. ConclusionThe effect of YSJD on significantly reducing the extent of HF in mice caused by CCl4 may be related to its ability to regulate liver hardness and inhibit the occurrence of epithelial mesenchymal transition in mice.

Effect of Sanhuang Tangshenkang on Wnt/β-catenin Signaling Pathway in Bone Tissue of Diabetic Rats / 中国实验方剂学杂志

ObjectiveTo observe the effect of Sanhuang Tangshenkang on the Wnt/β-catenin signaling pathway in the bone tissue of diabetic rats. MethodA high-sugar and high-fat diet was administered for 4 weeks， along with intraperitoneal injection of freshly prepared 2% streptozotocin （pH 4.5） at 30 mg·kg-1 body weight to induce a diabetes model in rats. The rats with diabetes were randomly divided into model group， low- and high-dose Sanhuang Tangshenkang groups （12.8， 38.4 g·kg-1）， and Gushukang group （1.8 g·kg-1） according to the blood glucose level. Rats of the same age were fed on a regular diet and assigned to the control group. After 12 weeks of respective treatments with drugs or physiological saline， the fasting blood glucose （FBG） levels of the rats were measured using an automated biochemical analyzer. Enzyme-linked immunosorbent assay （ELISA） was used to detect fasting serum insulin （FINS）， bone-specific alkaline phosphatase （BALP）， and tartrate-resistant acid phosphatase （TRAP） levels. Micro-computed tomography （Micro-CT） was used to scan the femurs of rats to observe bone tissue microstructure and measure bone mineral density （BMD）. Hematoxylin-eosin （HE） staining and safranin O/fast green staining were performed to observe pathological changes in the femoral bone tissue. Immunohistochemistry （IHC） and Western blot were used to detect the expression of Wnt3a， low-density lipoprotein receptor-related protein 5 （LRP-5）， and β-catenin proteins. ResultCompared with the control group， the model group showed a significant increase in FBG， FINS， and TRAP levels （P<0.01）， a significant decrease in BALP level （P<0.01）， a significant decrease in BMD （P<0.01）， and disorganized， elongated， and sparse bone trabecular structures with fractures and increased lipid droplets. Additionally， the expression of Wnt3a， LRP-5， and β-catenin proteins decreased （P<0.05， P<0.01）. Compared with the model group， the low- and high-dose Sanhuang Tangshenkang groups showed a reduction in FBG and an increase in BALP （P<0.05）. The low-dose Sanhuang Tangshenkang group also exhibited a decrease in FINS （P<0.05）. All treatment groups showed a significant decrease in TRAP （P<0.01）， varying degrees of improvement in BMD （P<0.05， P<0.01））， increased and denser bone trabeculae with more regular arrangements and reduced lipid droplets， and improved bone microstructure morphology. The average optical density values of Wnt3a， LRP-5， and β-catenin proteins were significantly increased in all drug-treated groups （P<0.05， P<0.01）， and the expression of Wnt3a， LRP-5， and β-catenin proteins was elevated （P<0.05， P<0.01）. ConclusionSanhuang Tangshenkang may regulate the imbalance of the Wnt/β-catenin signaling pathway by increasing the expression of Wnt3a， LRP-5， and β-catenin proteins in bone tissue， which may promote bone formation， reduce bone resorption， and lower blood glucose levels， thereby achieving the effect of preventing and treating diabetic osteoporosis.

Dong's extraordinary point needling technique combined with medication for postoperative complications of anal fistula: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the effect of Dong's extraordinary point needling technique on postoperative complications of anal fistula.@*METHODS@#A total of 241 patients undergoing anal fistula surgery were randomly divided into an observation group (121 cases, 3 cases dropped off) and a control group (120 cases, 2 cases dropped off). The patients in the control group were treated with intramuscular injection of compound diclofenac sodium injection and oral administration of tamsulosin hydrochloride sustained release capsules. In addition to the treatment in the control group, the patients in the observation group were treated with Daoma needling technique at the "Sanqi points" (Qimen point, Qijiao point, and Qizheng point) combined with Dongqi needling technique at "Sanhuang points" (sub-Tianhuang point, Dihuang point, Renhuang point), with each session lasting 30 min. The treatment in the two groups both started on the first day after surgery, and was given once daily for 14 consecutive days. Visual analog scale (VAS) score was compared between the two groups on postoperative day 1, 7, and 14; bladder residual urine volume, spontaneous voiding volume, and urinary catheterization frequency were assessed after treatment on postoperative day 1; and anorectal dynamic indexes (anal canal resting pressure, rectal resting pressure, maximum squeeze pressure of the anal canal, and minimum rectal sensory threshold) were evaluated before surgery and on postoperative day 4. Clinical efficacy was assessed in both groups one month after surgery.@*RESULTS@#On postoperative day 7 and 14, the VAS scores of both groups were lower than those on postoperative day 1 (P<0.05), and the VAS scores in the observation group were lower than those in the control group (P<0.05). The bladder residual urine volume and urinary catheterization frequency in the observation group were lower than those in the control group (P<0.05), while the spontaneous voiding volume was higher than that in the control group (P<0.05). On postoperative day 4, the anal canal resting pressure, maximum squeeze pressure of the anal canal, and the minimum rectal sensory threshold were lower than preoperative values (P<0.05), while the rectal resting pressure was higher than preoperative value (P<0.05) in both groups. The anal canal resting pressure, maximum squeeze pressure of the anal canal, and minimum rectal sensory threshold were lower than those in the control group, and the rectal resting pressure was higher than that in the control group (P<0.05). The effective rate was 93.2% (110/118) in the observation group, which was higher than 84.7% (100/118) in the control group (P<0.05).@*CONCLUSION@#Dong's extraordinary point needling technique could reduce postoperative pain, alleviate urinary retention, and improve defecation in patients undergoing anal fistula surgery.

Pharmacodynamic mechanism study on Modified sanhuang ointment in the treatment of anal ulcers and swelling in rats / 中国药房

OBJECTIVE To explore the improvement mechanism of Modified sanhuang ointment on anal ulcers and swelling model rats. METHODS The anal ulcer model of the rat was induced by using glacial acetic acid； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on body weight， area of anal ulcer， grade of anal ulcer were investigated. The other groups of rats were used to induce rectal swelling models with croton oil； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on the rate of rectal and anal swelling， serum contents of inflammatory factors [interleukin 2 （IL-2）， IL-4， tumor necrosis factor （TNF-α）]， pathological morphology of rectal tissue， the expression of transient receptor potential channel V1 （TRPV1） and substance P in rectal tissue and rectal vascular permeability were investigated. RESULTS In Modified sanhuang ointment， the increase in body weight was enhanced， and the area of anal ulcers， as well as the grade of anal ulcers in rats with anal ulcer models， were reduced to varying degrees； rectal tissue damage in rectal swelling model rats was improved； the rate of rectal and anal swelling， the serum contents of inflammatory factors， the expressions of TRPV1 and substance P in rectal tissue， and rectal vascular permeability were all decreased （P＜0.05 or P＜0.01）. The effect of Modified sanhuang ointment was better than that of western medicine positive control and TCM positive control.Modified sanhuang ointment can improve anal ulcers and swelling in rats by reducing the release of inflammatory factors， inhibiting the expression of TRPV1 and substance P.

Clinical study on Sanhuang Qingre Fuming and Washing Decoction combined with Chinese medical excision in the treatment of horseshoe perianal abscess after high position / 国际中医中药杂志

Objective:To observe the clinical effect of Sanhuang Qingre Fuming and Washing Decoction combined with Chinese medical excision in the treatment of horseshoe-shaped perianal abscess after high position and to explore its mechanism of action.Methods:Randomized controlled trial. Sixty surgical patients in our hospital from April 2020 to February 2021 were selected as observation objects. The patients were divided into two groups according to the random number table, with 30 patients in each group. Both groups were treated with Traditional Chinese Medicine drainage. On the second day after operation, the control group was treated with potassium permanganate sitz bath fumigation and washing, and the observation group was treated with Sanhuang Qingre Fuming and Washing Decoction. The wound healing time was observed in the two groups, serum TNF-α, IL-1β, IL-6, VEGF, bFGF, and EGF levels were detected by ELISA on the 1st and 7th d after operation, respectively. Serum SOD level was detected by the chemiluminescence method, and serum MDA level was detected by thiobarbituric acid method.Results:The wound healing time in the observation group was shorter than that in the control group ( t=3.37, P<0.01). The recovery rate was 93.3% (28/30) in the observation group and 76.7% (23/30) in the control group. The difference was statistically significant between the two groups ( χ2=4.32, P=0.038). Seven days after surgery, the levels of serum TNF-α, IL-1β and IL-6 in the observation group were significantly lower than those in the control group ( t values were 2.01, 2.05 and 4.88, respectively, P<0.05 or P<0.01), the level of SOD was significantly higher than that of the control group ( t=-2.13, P=0.038), the level of MDA water was significantly lower than that of the control group ( t=3.26, P=0.002), and the levels of VEGF, bFGF and EGF were significantly higher than those in the control group ( t values were -2.02, -3.33, -3.77, respectively, P<0.05). Conclusion:Sanhuang Qingre Tuming and Washing Decoction combined with Chinese medical excision in the treatment could shorten the healing time of the wound surface of horseshoe-shaped perianal abscess after high elevation and improve the recovery rate. The Sanhuang Qingre Fuming and Washing Decoction may reduce inflammatory reaction and oxidative stress damage, improve the level of growth factors and facilitate the postoperative tissue repair.

Mechanism of Sanhuang Decoction in alleviating dextran sulfate sodium induced ulcerative colitis in mice with Candida albicans colonization:based on high-throughput transcriptome sequencing / 中国中药杂志

This study explored the mechanism of Sanhuang Decoction(SHD) in treating dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice with Candida albicans(Ca) colonization via high-throughput transcriptome sequencing. Specifically, the animal model was established by oral administration of 3.0% DSS for 7 days followed by intragastrical administration of Ca suspension at 1.0 × 10~8 cells for 4 days and then the mice were treated with SHD enema for 7 days. Afterwards, the general signs were observed and the disease activity index(DAI) was recorded every day. After mice were sacrificed, colon length and colon mucosa damage index(CMDI) were determined and the histomorphology was observed with the HE staining method. The fungal loads of feces were detected with the plate method. Anti-saccharomyces cerevisiae antibody(ASCA) and β-1,3-glucan in serum, and TNF-α, IL-1β, and IL-6 in serum and colon were detected by ELISA. High-throughput RNA sequencing method was adopted to identify transcriptome of colon tissues from the control, model and SHD(15.0 g·kg~(-1)) groups. Differentially expressed genes(DEGs) among groups were screened and the GO and KEGG pathway enrichment analysis of the DEGs was performed. The expression levels of NLRP3, ASC, caspase-1, and IL-1β genes related to the NOD-like receptor signaling pathway which involved 9 DEGs, were examined by qRT-PCR and Western blot. The results demonstrated that SHD improved the general signs, decreased DAI and Ca loads of feaces, alleviated colon edema, erosion, and shortening, and lowered the content of β-1,3-glucan in serum and TNF-α, IL-1β, and IL-6 in serum and colon tissues of mice. Transcriptome sequencing revealed 383 DEGs between SHD and model groups, which were mainly involved in the biological processes of immune system, response to bacterium, and innate immune response. They were mainly enriched in the NOD-like signaling pathway, cytokine-cytokine interaction pathway, and retinol metabolism pathway. Moreover, SHD down-regulated the mRNA and protein levels of NLRP3, caspase-1, and IL-1β. In a word, SHD ameliorates DSS-induced UC in mice colonized with Ca, which probably relates to its regulation of NOD-like receptor signaling pathway.

Effect of Shenteng Sanhuang Decoction and Gegen Qinlian Tang on Diabetic Kidney Disease Rats / 中国实验方剂学杂志

Objective::To explore the protective mechanism of Shenteng Sanhuang decoction and Gegen Qinlian Tang on diabetic nephropathy (DN). Method::Rat DN models were duplicated with unilateral nephrectomy combined with streptozotocin (STZ). The rats were randomly divided into blank group, model group, irbesartan group and traditional Chinese medicine group. After 8 weeks of administration of corresponding drugs, the body weight, blood sugar, blood urea nitrogen (BUN), 24-hour urine volume (24 h U-vol), 24-hour urinary protein (24 h U-pro), serum creatinine (SCr), kidney weight/body weight (KW/BW) mass index of rats in each group were measured. The kidney tissues of rats in each group were homogenized, and supernatant was taken. Expressions of Smad ubiquitination regulatory factor 2 (Smurf2), matrix metalloprotein-9 (MMP-9), TGF-β1 and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were detected by Western blot or special test kit. Result::Compared with the blank group, the biochemical indicators, body weight, KW/BW, blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly higher or increased(P<0.05), while SOD and CAT were significantly decreased in the model group(P<0.05). Western blot showed that the expression of Smurf2 and TGF-β1 was high, while the expression of MMP-9 was low(P<0.05). Compared with the model group, the biochemical indicators of irbesartan group and traditional Chinese medicine group improved significantly(P<0.05), KW/BW were reduced, and blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly decreased (P<0.05), SOD and CAT was obviously increased (P<0.05), expressions of Smurf2 and TGF-β1 were decreased significantly(P<0.05), and expression of MMP-9 was increased significantly(P<0.05). Conclusion::Shenteng Sanhuang decoction and Gegen Qinlian Tang can effectively improve many biochemical indexes of rat DN models, and improve renal function. Its mechanism is closely related to reducing the expressions of Smurf2 and TGF-β1, and enhancing the expression of MMP-9.

Effect of Sanhuang Xiexintang in Inhibiting 7-Ketocholesterol-induced Endothelial Disfunctional by NIRP3 Inflammasome Pathway / 中国实验方剂学杂志

<b>Objective::To study whether Sanhuang Xiexintang (SHXXT) can restore endothelial function by inhibiting the activation of NOD-like receptor protein 3 (NIRP3) induced by 7-ketocholesterol (7-keto) in vascular endothelial cells. <b>Method::The aortic rings of mice were cultured in normal group, model (7-keto) group, SHXXT groups (1%, 2% and 5% drug-containing serum). Vasodilation function of mice was observed. Microvascular endothelial cells were cultured according to the above experimental groups, and NIRP3 inhibitor isoglycyrrhizin (ISO) group, was also set. Western blot was used to detect the expressions of endothelial nitric oxide synthase (eNOS), NIRP3, cysteinyl aspartate specific proteinase-1 (Caspase-1), interleukin-1<italic>β</italic> (IL-1<italic>β</italic>) protein. In addition, nitric oxide (NO) quantitative kit was used to detect the concentration of NO. <b>Result::Compared with the normal group, the endothelium-dependent vasodilation function of vascular rings was significantly reduced in model group (<italic>P</italic><0.01), and the drug group significantly restored the endothelium-dependent vasodilation function in a concentration-dependent manner (<italic>P</italic><0.05, <italic>P</italic><0.01). Meanwhile, microvascular endothelial cells were also studied. Compared with the normal group, the content of eNOS protein in the model group decreased (<italic>P</italic><0.05), while the concentration of NO decreased significantly (<italic>P</italic><0.01). After treatment with SHXXT serum, eNOS and NO could be restored, with significant differences in the concentration of NO with 5% (<italic>P</italic><0.05) and 10% (<italic>P</italic><0.01) SHXXT serum. At the same time, the expressions of NIRP3 (<italic>P</italic><0.05), cle-Caspase-1 activation (<italic>P</italic><0.01) and IL-1<italic>β</italic> production (<italic>P</italic><0.01) in endothelium were significantly increased under 7-keto stimulation, and the SHXXT serum could significantly inhibit the expression and activation of relevant proteins. Subsequently, endothelial cells were treated with NIRP3 inhibitor ISO. Compared with the model group, eNOS expression increased, and NO concentration increased significantly (<italic>P</italic><0.01) after treatment with ISO, but ISO had no synergistic effect on SHXXT serum. <b>Conclusion::SHXXT can improve endothelium-dependent vascular dysfunction induced by 7-keto, which is achieved by NO signaling pathway mediated by inhibiting the activation of endothelial NIRP3-related proteins.

Clinical treatment of chronic nonbacterial prostatitis of kidney-yang deficiency type by acupuncture of Sanhuang points / 针刺研究

OBJECTIVE: To observe the clinical effect of acupuncture stimulation of Sanhuang points in the treatment of chronic nonbacterial prostatitis (CNP) of kidney-yang deficiency type. METHODS: A total of 60 CNP outpatients were equally randomized into medication(control) group and acupuncture plus medication (acupuncture) group. Patients of the control group were ordered to take Tamsulosin (0.2 mg/d) for successive 8 weeks. On the basis of medication treatment, for patients of the acupuncture group, bilateral Sanhuang points including Tianhuang-fu Point (Shenguan), Dihuang Point and Renhuang Point (at the lower leg) were needled with filiform needles which were manipulated for a while till Deqi, followed by retaining the needles for 30 min. The treatment was conducted once daily for successive 8 weeks. The therapeutic effect was evaluated in reference to the "Criteria for Diagnosis and Therapeutic Effect Evaluation of Syndromes/Diseases of Traditional Chinese Medicine". The National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) scores were used to assess the severity of chronic prostatitis. Routine examination of the leukocyte count in the prostatic fluid was performed before and after the treatment. RESULTS: Of the two 30 cases of the control and acupuncture groups, 9 and 19 were cured, 14 and 10 experienced improvement, and 7 and 1 failed in the treatment, with the effective rates being 76.67% and 96.67%, respectively. The effective rate of the acupuncture plus medication was significantly higher than that of the medication (P<0.05). After the treatment, the scores of NIH-CPSI and white blood cell counts in the prostatic fluid in both groups were significantly decreased in comparison with their own pre-treatment (P<0.05). The therapeutic effect of acupuncture plus medication was significantly superior to that of the medication alone in reducing the NIH-CPSI symptom score and the number of leukocytes in the prostatic fluid (P<0.05). CONCLUSION: Acupuncture at Sanhuang points is effective in improving symptoms of CNP patients of kidney-yang deficiency type, and the therapeutic effect of acupuncture plus medication is superior to that of medication alone.

Mechanism of Modified Taohe Chengqitang in Treating Diabetic Nephropathy Based on Network Pharmacology / 中国实验方剂学杂志

Objective: To study the mechanism of modified Taohe Chengqitang in preventing and treating diabetic nephropathy by means of network pharmacology. Method: Target genes of modified Taohe Chengqitang were obtained from BAT-MAN database, while target genes of diabetic nephropathy were obtained from CTD database. The target genes of disease-drug protein were obtained by crossing two groups of genes. STRING was used to build the protein-protein interaction network and visualize the results. The key genes were screened out through the computational analysis algorithm of network structure and weighted relatedness between nodes. With DAVID online tools, gene ontology (GO) analysis of Disease-Drug Intersection Target Genes and enrichment analysis of kyoto encyclopedia of genes and genomes(KEGG) pathway were conducted. Finally, CTD database and literature study were used to obtain the key genes in the treatment of diabetic nephropathy. Result: Among 621 compounds in modified Taohe Chengqitang, 581 of them were related to diabetic nephropathy. NOS3, OAT, NT5C2, ACACB, AGXT, PDE3B and other key genes mainly regulated nerve tissue transmission, cholinergic synaptic pathway, calcium channel, metabolic pathway, purine metabolic pathway, angiotensin-neurosynaptic pathway, cyclic guanosine monophosphate/cGMP-dependent protein kinase G (cGMP/PKG) signaling pathway and cyclic adenosine phosphate signaling pathway, with effect in molecular reactions, such as plasma membrane, postsynaptic membrane and mitochondria. Conclusion: The network pharmacology predicts the key targets of modified Taohe Chengqitang in the prevention and treatment of diabetic nephropathy and the related pathways involved, suggesting a multi-target, multi-channel and multi-choice complex mechanism, and which is mostly related to anti-inflammation, oxidative phosphorylation and purine metabolism.

Effect of Sanhuang Yinchi Decoction in Preventing Acute Liver Injury by HMGB1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of Sanhuang Yinchi decoction (SHYCD) in preventing carbon tetrachloride (CCl4)-induced acute liver injury by regulating high mobility group box1(HMGB1) signaling pathway. Method:A total of 48 KM mice were randomly divided into blank control group, model group, low, middle and high-dose SHYCD groups and positive control group. The model of acute liver injury induced by CCl4 in mice was established. The low, middle and high-dose SHYCD groups were intragastrically administered with drugs (16, 32, 48 g·kg-1·d-1) respectively, and the positive control group was given cell growth stimulating hormone (20 mg·kg-1·d-1) through intraperitoneal injection. Pathological changes of mouse liver tissue sections were observed by hematoxylin-eosin staining (HE); relevant enzyme kits were used to determine liver function indexes in mice serum-alanine aminotransferase (AST) and aspartate aminotransferase (ALT); the expression level of interleukin-6 (IL-6) in mouse serum was determined by enzyme-linked immunosorbent assay (ELISA); Western blot was used to detect the expressions of high mobility group box-1(HMGB1), cysteine aspartic acid protease(Caspase-3), apoptosis-related molecules B cell lymphoma(Bcl-2), Bcl-2 associated x protein(Bax), and Toll-like receptor 4 (TLR4). Result:Compared with the normal group, the model group significantly increased serum AST, ALT (PPPPPConclusion:SHYCD can prevent liver injury by regulating HMGB1/TLR4/NF-κB signaling pathway, reducing cellular inflammatory response and inhibiting apoptosis, so as to prevent acute liver injury in mice. This indicates that HMGB1 may become a new target to prevent acute liver injury.

Simultaneous determination of four components in Sanhuang tablets by RP-HPLC / 国际中医中药杂志

Objective To establish a RP-HPLC method for simultaneously determining berberine hydrochloride, baicalin, emodin and chrysophanol in Sanhuang tablets. Methods The analysis was performed on an Thermo Hypersil GOLD aQ column (250 mm×4.6 mm, 5.0 μm) with gradient elution of methanol and 0.1％orthophosphoric acid at the flow rate of 1.0 ml/min. The column temperature was 30 , and the detection wave℃length was 254 nm. Results The peaks of berberine hydrochloride, baicalin, emodin and chrysophanol were successfully separated from each other. The liner ranges of calibration curves were 0-0.6348 μg (r=0.9999), 0-1.0172 μg (r=0.9999), 0-0.8024 μg (r=0.9999), 0-0.9672 μg (r=0.9999). The average recoveries (n=9) were 101.03％, 99.81％, 99.35％, 99.54％, respectively. The limits of quantification (LOQs) were 0.0363, 0.0210, 0.0497, 0.0793 mg/g, respectively. The sample solution was stable within 24 h, the RSD (n=5) were 0.48％, 0.56％, 0.68％, 0.49％, respectively. The robustness was investigated by varying the conditions of column temperature (± 1 ℃) and flow rate (± 0.1 ml/min), with the RSD s 0.57％ and 0.42％, 0.44％ and 0.65％, 0.74％ and 0.29％, 0.46％ and 0.56％, respectively. The contents of berberine hydrochloride, baicalin, emodin and chrysophanol in twelve samples were 4.6303-5.5866, 14.5514-18.8189, 0.1711-2.7984, 0.5299-2.9254 mg/tablet, respectively. Conclusion The establish method is validated to be suitable for quality control of Sanhuang tablets.

An Outbreak of Histomoniasis in Backyard Sanhuang Chickens

Histomonas meleagridis is a facultative anaerobic parasite, which can cause a common poultry disease known as histomoniasis. The species and age of the birds impacts on the susceptibility, with turkey being the most susceptible species. Chickens are less susceptible to H. meleagridis than turkeys and usually serve as reservoir hosts. Here, the diagnosis of an outbreak of histomoniasis in backyard Sanhuang chickens is described. The primary diagnosis was made based on clinical symptoms, general changes at necropsy, histopathology, and the isolation and cultivation of parasites. The pathogen was further confirmed by cloning, PCR identification, and animal inoculation tests. A strain of H. meleagridis, named HM-JSYZ-C, with a higher pathogenicity level in chickens was obtained. The study lays a foundation for further investigations into H. meleagridis and histomoniasis in chickens.

Effects of Different Dose Compatibility of Scutellaria baicalensis-Rheum palmatum-Coptis chinensis Based on Sanhuang Xiexin Decoction on Dissolution Rate of Baicalin and Baicalein / 中国药房

OBJECTIVE: To investigate the effects of different dose compatibility of Scutellaria baicalensis-Rheum palmatum-Coptis chinensis based on Sanhuang Xiexin decoction (shorted for Xiexin decoction) on dissolution rate of baicalin and baicalein, and to provide reference for studying chemical mechanism of TCM compound dose composition compatibility. METHODS: HPLC method was used to determine the contents of baicalin and baicalein. By fixing the dose of S. baicalensis (3 g), using the dose of R. palmatum and C. chinensis as factors, dissolution rate of baicalin and baicalein as response value, two-factor and five-level central composite test was designed. The optimal dose compatibility of S. baicalensis-R. Palmatum-C. chinensis was optimized by response surface method, and compared with the dissolution rate of baicalin and baicalein in classic dose Xiexin decoction (S. baicalensis 3 g, R. Palmatum 6 g, C. chinensis 3 g). RESULTS: When the doses of S. baicalensis, R. palmatum, C. chinensis were 3, 1. 76, 0. 17 g, total dissolution rate of baicalin and baicalein was the highest in extract. The average total dissolution rate of baicalin and baicalein in validation test was 21. 89％ (RSD=0. 46％, n=3),and the relative error was 2. 88％ with the predicted value of 22. 54％. Compared with the classical dose of Xiexin decoction, the total dissolution rate of baicalin and baicalein in the optimum dose of S. baicalensis-R. Palmatum-C. chinensis in extract was increased by 47. 21％. CONCLUSIONS: The different dose compatibility of S. baicalensis-R. Palmatum-C. chinensis based on Xiexin decoction influence the dissolution of baicalin and baicalein to certain extent. When the doses of S. baicalensis, R. palmatum, C. chinensis are 3, 1. 76, 0. 17 g, dissolution rate of baicalin and baicalein are higher than that of classic dose.

Clinical observation of Sanhuang-Jianchi decoction combined with routine treatment of western medicine for treatment of patients with chronic periodontitis / 国际中医中药杂志

Objective To explore the clinical curative effect and safety of Sanhuang-Jianchi decoction combined with routine treatment of western medicine in treatment of patients with chronic periodontitis. Methods According to the randomindicator method, 83 patients with chronic periodontitis conformed to the standard of admission were divided into control group (n=41) and treatment group (n=42). Patients in the control group were treated with periodontal basal treatment and tinidazole, while the treatment group was treated with Sanhuang-Jianchi decoction on the base of control group. The two groups of patients were treated for one month and followed-up for three months. The probing depth (PD), attachment loss (AL) and gingival bleeding index (BI) of two groups of patients were observed before and after treatment. At the same time, the TCM symptom score and gingival sulcus fluid tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and IL-8 were detected before and after the treatment. In addition, the incidence of adverse reactions and the recurrence rate in both groups were observed. Results The total clinical effectiveness of the treatment group was 95.2％ (40/42) and the control group was 75.6％ (31/41). The difference of two groups was statistically significant (χ2=4.363, P=0.037). After treatment, the PD (2.37 ± 0.22 mm vs. 3.04 ± 0.19 mm, t=-14.834), AL (2.53 ± 0.95 mm vs. 3.61 ± 1.05 mm, t=-4.916), BI (1.43 ± 0.13 vs. 2.05 ± 0.17, t=-18.692), the score of ozostomia (1.09 ± 0.51 vs. 1.92 ± 0.64, t=-6.542), Bleeding gums (1.28 ± 0.62 vs. 2.03±0.56, t=-5.779), dry mouth (1.46 ± 0.512 vs. 2.01 ± 0.56, t=-4.680), constipation (1.13 ± 0.47 vs. 1.89 ± 0.54, t=-6.844), tooth non-traumatic (1.06 ± 0.42 vs. 1.73 ± 0.52, t=-6.465), yellow unary (1.17 ± 0.37 vs. 1.81 ± 0.65, t=-5.529) in the treatment group were significantly lower than the control group. The level of TNF-α (3.05 ± 1.02 ng/ml vs. 4.22 ± 1.04 ng/ml, t=-5.174), IL-1β (3.29 ± 1.02 pg/ml vs. 6.24 ± 1.35 pg/ml, t=-11.250), IL-6 (4.15 ± 1.13 pg/ml vs. 6.02 ± 1.24 pg/ml, t=-7.184) and IL-8 (4.03 ± 1.02 pg/ml vs. 6.25 ± 1.19 pg/ml, t=-9.133) of treatment group were significantly lower than control group.The recurrence rate was 5.0％ (2/40) in treatment group and 22.6％ (7/31) in the control group during the 3 months of follow-up, and the difference was statisticlly significant (χ2=4.877, P=0.027). Conclusions The Sanhuang-Jianchi decoction combined with routine treatment of western medicine can improve the clinical symptoms, restrain the inflammation of gingival sulcus of liquid and reduce recurrence rate.

Simultaneous determination of nine constituents in Sanhuang Tablets by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of berberine hydrochloride,baicalin,baicalein,wogonin,aloe emodin,rhein,emodin,chrysophanol,physcion in Sanhuang Tablets (Rhei Radix et Rhizoma,Scutellariae Radix,berberine hydrochloride).METHODS The analysis of methanol extract of this drug was performed on a 35 ℃ thermostatic Waters sunfire C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.RESULTS Nine constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 99.03％-100.03％ with the RSDs of 0.63％-1.07％.The contents of various constituents in ten batches of samples demonstrated obvious differences.CONCLUSION We should pay attention to the unstable quality of Sanhuang Tablets.

Optimization of Extraction Process of Berberine Hydrochloride from Sanhuang Diyu Oil by Box-Benhnken Response Surface Methodology / 中国药师

Objective:To optimize the extraction process of berberine hydrochloride from Sanhuang Diyu oil by Box-Benhnken re-sponse surface methodology. Methods:The extraction process was optimized with the size of each medicine ( X1 ) , the liquid-solid ra-tio ( X2 ) and the extraction time ( X3 ) as the independent variables and berberine hydrochloride extraction amount ( Y) as the depend-ent variable, and the effects of variables and their interactions on the extraction efficiency were studied. Results:The experiment results indicated that the optimal extraction conditions were as follows:the size of each medicine was 60 meshes, the liquid-solid ratio was 18, and the extraction time was 1. 5 h. Under the above conditions, 3 batches of berberine hydrochloride from Sanhuang Diyu oil were ex-tracted. The average extraction rate of berberine hydrochloride was (16.6 ±0.6) mg·g-1(n =3). Conclusion: The extraction process of Sanhuang Diyu oil optimized by Box-Behnken response surface methodology is simple, stable and predictable.