Polydatin Ameliorated Colonic Inflammation via Precluding Phosphorylation of PKCθ/STAT3 Pathway to Inhibit Th17 Cells in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of polydatin on ulcerative colitis （UC） in mice and its regulation of protein kinase C<italic>θ</italic>（PKC<italic>θ</italic>）/signal transducer and activator of transcription 3（STAT3） signaling on T helper cell 17（Th17） and its mechanism in the treatment of UC. Method:The 32 male C57BL/6 mice were randomly divided into normal group， model group， polydatin group （0.045 g·kg^-1） and sulfasalazine group （0.5 g·kg^-1）. The UC model was established by giving 3% dextran sodium sulfate （DSS） solution to free drinking water in mice. Polydatin and sulfasalazine groups were given by gavage 0.5 h before modeling for 7 days. The normal group and model group were given the same amount of normal saline. After the last administration， the colonic tissue was taken and hematoxylin-eosin （HE） was used to observe the pathological changes of colonic tissue. Flow cytometry was used to detect the proportion of Th17 in the lamina propria of colonic mucosa. The expression of interleukin-17A （IL-17A） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. Polydatin was added to CD4⁺T cells purified from spleen of C57BL/6 mice by magnetic-activated cell sorting （MACS） under the stimulation of cell stimulation cocktail <italic>in vitro </italic>in order to detect its impact on PKC<italic>θ</italic> and STAT3 phosphorylation. Result:Compared with normal group， the body weight was significantly decreased， and disease activity index （DAI） scores of the model group was significantly increased （<italic>P</italic><0.01）， the colonic mucosal epithelium was damaged and inflammatory cells infiltration in the mucosa and submucosa was obvious， the proportion of Th17 in the lamina propria of colonic mucosa was significantly increased （<italic>P</italic><0.01）， and the content of serum IL-17A was significantly increased （<italic>P</italic><0.01）. Compared with the model group， the weight and DAI score of polydatin and sulfasalazine groups were significantly improved （<italic>P</italic><0.01）， the degree of colon tissue damage was significantly improved， the proportion of Th17 in colon mucosa lamina propria was significantly decreased （<italic>P</italic><0.01）， and the content of IL-17A in serum was significantly decreased （<italic>P</italic><0.01）. <italic>In vitro</italic> experiments showed that polydatin could significantly inhibit the phosphorylation of PKC<italic>θ</italic> and STAT3 in Th17 （<italic>P</italic><0.01） as well as IL-17A secretion. Conclusion:Polydatin can improve the ulcerative colitis in mice via inhibiting the phosphorylation of PKC<italic>θ</italic> and STAT3 to preclude IL-17A secreting in Th17.

Gegen Qinliantang Regulates MMP-9/p38 MARK Pathway to Repair Intestinal Mucosal Barrier Function in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective:To explore the protective effect of Gegen Qinliantang on the intestinal mucosal epithelial barrier function of ulcerative colitis （UC） mice， and to explore its mechanism of action in the treatment of ulcerative colitis via matrix metallopeptidase-9 （MMP-9）/p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway. Method:The 48 female C57BL/6 mice were randomly divided into normal group， model group， sulfasalazine group （0.3 g·kg^-1） and Gegen Qinliantang high， medium and low dose groups （2.84，1.42，0.71 g·kg^-1）. The UC murine model was established by 3% dextran sulfate sodium （DSS）. Gegen Qinliantang and sulfasalazine were intragastrically administered on the 8^th day after the model was established for 7 days， and the normal group was treated with the same amount of normal saline. Colon tissues were collected after the last administration， and the pathological changes of colon tissues were detected by hematoxylin-eosin （HE） staining. The expression of tight junction （TJ） proteins such as Occludin and zonula occludens-1（ZO-1） in colon tissues was detected by immunohistochemistry （IHC）， and the expression levels of tumor necrosis factor-alpha （TNF-<italic>α</italic>）， interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， and MMP-9 mRNA in colon tissues were detected by Real-time polymerase chain reaction （Real-time PCR）. The expression of phosphorylated p38 MAPK （p-p38 MAPK）， p38 MAPK and MMP-9 protein in colon tissues was detected by Western blot. Result:Compared with normal group， the body weight of mice decreased （<italic>P</italic><0.01） and disease activity index （DAI） score increased significantly （<italic>P</italic><0.01） in model group， the colon tissues of the model group were damaged more obviously， the expression of occludin and ZO-1 proteins in model group was significantly reduced （<italic>P</italic><0.01）， and the relative expression levels of TNF-<italic>α</italic>， IL-1<italic>β</italic>， and MMP-9 mRNA in model group were significantly increased （<italic>P</italic><0.01）， the expression of p-p38 MAPK and MMP-9 in model group was significantly increased （<italic>P</italic><0.01）. Compared with model group， the body mass and DAI score of the sulfasalazine group and Gegen Qinliantang group were significantly improved （<italic>P</italic><0.05，<italic>P</italic><0.01）， the colonic tissues damage were significantly improved， and the expression of Occludin and ZO-1 protein was significantly increased （<italic>P</italic><0.05，<italic>P</italic><0.01）， the relative expression levels of TNF-<italic>α</italic>， IL-1<italic>β</italic>， and MMP-9 mRNA were significantly decreased （<italic>P</italic><0.01）， and the expression of p-p38 MAPK and MMP-9 was significantly decreased （<italic>P</italic><0.01）. The changes in the middle dose group were the most obvious among the various dose groups of Gegen Qinliantang. Conclusion:Gegen Qinliantang repairs the intestinal mucosal barrier function by inhibiting the expressions of MMP-9 and inflammatory cytokines such as TNF-<italic>α</italic> and IL-1<italic>β</italic>， blocking the activation of the p38 MAPK signaling pathway， and increasing the expressions of tight junction protein.

Serum metabolic profiling of traditional Chinese medicine syndromes in patients with diarrhea-predominant irritable bowel syndrome / 中西医结合学报

OBJECTIVE@#The clinical symptoms of diarrhea-predominant irritable bowel syndrome (IBS-D) can be effectively improved by traditional Chinese medicine (TCM) treatment, based on the usage of specific therapies for different TCM syndromes. However, in the stage of diagnosis, the standard criteria for the classification of TCM syndrome were still deficient. Through serum metabolic profiling, this study aimed to explore potential biomarkers in IBS-D patients with different TCM syndromes, which can assist in diagnosis of the disease.@*METHODS@#Serum samples were collected from healthy controls (30 cases), IBS-D patients with Liver-Stagnation and Spleen-Deficiency syndrome (LSSD, 30 cases), Yang Deficiency of Spleen and Kidney syndrome (YDSK, 11 cases) and Damp Abundance due to Spleen-Deficiency syndrome (DASD, 22 cases). Serum metabolic profiling was conducted by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. The potential biomarkers were screened by orthogonal partial least square-discriminate analysis, while metabolic pathways undergoing alterations were identified by pathway enrichment analysis in MetaboAnalyst 4.0.@*RESULTS@#Overall, 34 potential biomarkers were identified in LSSD group, 36 in YDSK group and 31 in DASD group. And the 13 metabolites shared by three groups were determined as the potential biomarkers of IBS-D. Glycerophospholipid metabolism was disturbed significantly in IBS-D patients, which may play a role in IBS-D through inflammation. What's more, three TCM syndromes have the specific potential biomarkers in glycerophospholipid metabolism.@*CONCLUSION@#The serum metabolomics revealed that different TCM syndrome types in IBS-D may have different metabolic patterns during disease progression and glycerophospholipid metabolism was one of the pathways, whose metabolism was disturbed differently among three TCM syndromes in IBS-D. Therefore, the specific potential biomarkers in glycerophospholipid metabolism of three TCM syndromes in IBS-D can serve as the objective indicators, which can facilitate the TCM-syndrome objective classification of IBS-D.

Treatment of nonalcoholic steatohepatitis by Jianpi Shugan Recipe: a multi-center, randomized, controlled clinical trial / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Chinese medicine (CM) intervention in the treatment of nonalcoholic steatohepatitis (NASH) from liver enzyme (ALT), imaging (the liver/spleen CT ratio) and syndrome scores, and to establish standard methods for diagnosis and therapeutic efficacy evaluation with characteristics of CM.</p><p><b>METHODS</b>A multi-center, stratified randomized, parallel controlled, blindness-method evaluated, superiority trial was performed. Totally 204 patients were randomly allocated into two groups, 102 patients in the experimental group (treated with CM) and 102 patients in the control group [treated with Western medicine (WM)]. The alanine aminotransferase (ALT), liver/spleen CT ratio, and clinical symptoms were observed in both groups.</p><p><b>RESULTS</b>Of the randomly allocated 204 cases from 4 hospitals, 3 patients were rejected, and 25 were lost. Totally 176 cases con- formed to the plan with complete follow-ups. After 3 months of treatment, syndrome scores and the improvement of partial clinical symptoms (fatigue and sallow complexion) were superior in the experimental group to those in the control group (P < 0.05). After 3 months of follow-up, the syndrome scores and improvement of partial clinical symptoms (fatigue and sallow complexion) were superior in the experimental group to those in the control group (P < 0.05). There was no statistical difference in improving liver enzymes or the liver/spleen CT ratio between the two groups (P > 0.05). There were 4 adverse reactions/adverse events in the two groups in the process of treatment, mainly covering drug-induced liver injury, diarrhea, and epigastric distension. Adverse reactions had nothing to do with CM treatment.</p><p><b>CONCLUSIONS</b>Jianpi Shugan Recipe had obvious efficacy in treatment of NASH. It could remove the liver fat and play a role in anti-inflammation and liver protection. It also could improve the indices of liver enzymes and the liver/spleen CT ratio effectively, which was superior to Polyene Phosphatidylcholine Capsule (PPC) in improving clinical symptoms, especially for such symptoms as fatigue and sallow complexion.</p>

Snail habitats detection in the marshland of Eastern Dongting Lake Area, based on China-Brazil Earth Resources Satellite-02B CCD data / 中华流行病学杂志

Objective To detect the snail habitats from the marshland of Eastern Dongting Lake Area, using the Remote Sensing (RS) and Geographic Information System (GIS) technology based on the China-Brazil Earth Resources Satellite-02B (CBERS-02B) CCD images. Methods According to the two typical traits of snail habitats in marshland including "water in summer and land in winter" and "no grass, no snails" , the "water in summer and land in winter" region and the vegetation coverage region were calculated by RS image processing respectively. The two regions mentioned above were then overlapped to confirm the snail habitats through comparing with the data from field survey under spatial overlapping of Arcgis as the last step. Results In Eastern Dongting Lake area, the "water in summer and land in winter" region and vegetation coverage region were predicted based on the formula normalized difference water index (NDWI) >0.01 and normalized difference vegetation index (NDVI) >0.36, respectively. The snail habitat was determined by theme overlay of the two regions said above.The agreement rate between the prediction and the geospatial data of field survey was 93.55% , which demonstrated the final results were credible and reliable. Conclusion CBERS-02B image could be used to detect the snail habitats and to monitor the changes of them, so as to find out the characteristics of distribution and the trends of diffusion. The snail index (discriminant 1 and 2) seemed to be suitable for the detection of snail habitats in the marshland of Lake area and used for the programs of snail control.

Impact of the changing water level on the variance of Oncomelania hupensis populations in Lake Area with general additive model / 中华流行病学杂志

Objective To explore the impact and regularity of snail population after changing of water level and to develop effective control and prediction programs. Methods A typical snail habitat closed to Changjiang county in Yueyang city, Hunan province was selected as the survey field.Data on water level and the changing trend of snail population during 2001-2009 including the average water level, maximum and minimum water levels, snail and infected snail densities,proportion of sampling frames with living and infected snails etc. were collected. The distribution of water level and snail indexes were described and a general additive model(GAM)for the relationships between these indexes were also fitted. Results Impacts of the average water level and the watered-out days in current year in the surveyed field on the snail density were statistically significant(P＜0.01), showing a quadratic curve association. The snail density remained at the lowest level when the average water level was at 25.0 m and the surveyed field watered-out days was 120. The average water level in current year and the field watered-out days in the past year showed statistically significant impacts on the infected snail density(P＜0.001), the proportion of sampling frames with infected snails(P＜0.05)and living snails(P value neared 0.05), presented a quadratic curve, a cubic curve or even a more complicated piecewise curve association. Both the infected snail density and the proportion of sampling frames with infected snails remained at the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 110, while the proportion of sampling frames with living snails kept the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 160. Conclusion The water level and the field watered-out days affected the development of snail population directly. The changing water level had an impact on snail population change, which mainly presented as nonlinear smooth function relation. Impact of the field watered out days on the infected snail density and the proportion of sampling frames with living snails and infected snails showed a hysteresis effect. The snail density was predicted to be retaining a high level when the water level was 24.0 m and the field watered-out days was 3 months. It had obvious advantages to fit the relationship of the changing water level and the snail indexes with a GAM which could get closer to the reality as well as easier to find and explain the potential associations and regulations.

Coma criterion and classification standard of mild and moderate traumatic brain injury in rats / 法医学杂志

OBJECTIVE@#To set up a classification standard of mild and moderate traumatic brain injury, for the purpose of reliable data comparison derived from different laboratories.@*METHODS@#Traumatic brain injury (TBI) in rats was prepared by using a metallic pendulum-striker device. After injury, five variable parameters including the time of apnea and the areflexia, time of corneal reflex, external auditory canal stung reaction, body-righting reflex and needling reaction were determined and scored by using rat coma criterion. These data were judged and classified into mild and moderate head injury by brain patho-anatomy changes. Then the data were used to set up a multivariate discriminate equation.@*RESULTS@#The distinguished probability of mild and moderate TBI according to actual direct measured value and the criterion were 88.9% and 91.9%, respectively.@*CONCLUSION@#This method is able to classify mild and moderate TBI in rats.

Effect of three TCM methods for activating blood circulation on early stage apoptosis in rats with chronic atrophic gastritis complicated precancerous lesion / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To comparatively study the effects of three TCM methods for activating blood circulation, i.e. in combined with resolving stasis (A), regulating qi (B) and supplementing qi (C), respectively, on early stage cell apoptosis in precancerous lesion of rats with chronic atrophic gastritis (CAG).</p><p><b>METHODS</b>Rat model of CAG with precancerous lesion was duplicated by insertion of spring in pylorus and gastric perfusion of high-salt hot paste; and the impact of treatment on cell apoptosis was determined using Annexin V/PI double labeled flow cytometry.</p><p><b>RESULTS</b>After being intervened for 12 weeks, the early stage cell apoptosis rate in the natural recovery group was significantly higher than that in the control group (P <0.01); while it lowered more significantly in the three groups receiving TCM therapeutic methods for activating blood circulation, showing significant difference compared with the natural recovery group (P <0.01).</p><p><b>CONCLUSION</b>Three therapies of activating blood circulation all show inhibitory action on the early stage apoptosis of precancerous lesion in CAG rats, which is possibly one of their action mechanisms for improving or reversing the precancerous lesion.</p>

Cell proliferation inhibitive and apoptosis promoting effects of sanchi extract on GES-1 cell after being transformed by MNNG / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect drug contained canine serum, prepared by gastric perfusion with Sanchi extract (SE), in inhibiting proliferation and promoting apoptosis of cultured precancerous gastric cells by cell culture.</p><p><b>METHODS</b>The precancerous model cells (MC) used in the experiment were prepared through transforming eternalized human gastric mucosa epithelial cells GES-1 by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG). After once gastric perfusion of SE extract to dogs, the canine serum gotten before and at different time points after medication was used for test. The inhibitory effect of the drug serum obtained at different time points on MC after acting for 72 hrs was detected by 3-(4,5)-dimethy thioazol-2-yl-2,5-diphenyl-tetrazoliumbromide (MTT) method to find the optimal time point for drug serum preparation, that were 2 hrs and 6 hrs after medication. Then the cell apoptosis promoting effect after acting for 72 hrs of the drug serum obtained at the optimal time points was determined by flow cytometry.</p><p><b>RESULTS</b>The drug serum obtained at 2-hr and 6-hr after medication showed the highest inhibitive effect on MC cells, reaching 45.3% and 42.4% respectively, as compared with the effect of blank serum, the difference was significant (P<0.01). They could evidently promote the MC cell apoptosis, the apoptosis rate also showed significant difference to that of the blank serum (P < 0.05). Under their action, the proportion of MC cells in G0/G1 phase was obviously decreased (P < 0.05) while that in the G2/M phase significantly increased (P <0.05). However, the change of cells in S phase was not uniform.</p><p><b>CONCLUSION</b>The drug contained canine serum gotten 2 hr and 6 hr after SE feeding shows the optimal MC proliferation inhibitive effect and significant apoptosis promoting effect. Besides, it could significantly decrease the proportion of MC cells in G0/G1 phase and significantly increase that in G2/M phase, this effect might be one of the mechanisms of ES in inhibiting MC cell proliferation and promoting its apoptosis.</p>