Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis / 中国结合医学杂志

OBJECTIVE@#To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.@*METHODS@#The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.@*RESULTS@#The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (P<0.05 or P<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (P<0.05 or P<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (P<0.01).@*CONCLUSION@#GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.

Meta-analysis and trial sequential analysis of Chaihuang Granules in treatment of upper respiratory tract infection in children / 中国中药杂志

This study aimed to evaluate the efficacy and safety of Chaihuang Granules in the treatment of upper respiratory tract infection in children. The databases such as CNKI, Wanfang, VIP, SinoMed, Cochrane Library, PubMed, EMbase, Web of Science, Chinese Clinical Trial Registry, and ClinicalTrials.gov were searched for randomized controlled trial(RCT) of Chaihuang Granules for the treatment of upper respiratory tract infection in children, and supplemented by manual searching of gray literature. Two investigators independently screened the literature, extracted data, and assessed the methodological quality. Meta-analysis was performed using RevMan 5.4 software, trial sequential analysis was conducted using TSA 0.9.5.10 Beta software, and evidence quality evaluation was carried out using GRADE profiler 3.6.1 software. Eighteen RCTs involving 2 459 patients(1 262 in the treatment group and 1 197 in the control group) were included. Meta-analysis showed that compared with conventional therapy alone, Chaihuang Granules significantly improved the total effective rate(RR=1.18, 95%CI[1.15, 1.22], P<0.000 01), reduced the disappearance time of symptoms/signs(MD=-1.39, 95%CI[-1.66,-1.12], P<0.000 01), improved cytokine levels(MD=-2.40, 95%CI[-3.80,-1.00], P=0.000 8), improved humoral immune levels(MD=0.75, 95%CI[0.60, 0.90], P<0.000 01), and reduced the recurrence rate(MD=-2.11, 95%CI[-2.98,-1.25], P<0.000 01). However, the incidence of adverse reactions was not increased(RR=0.94, 95%CI[0.59, 1.49], P=0.78). Subgroup analysis showed that:(1) both Chaihuang Granules used alone(RR=1.19, 95%CI[1.11, 1.27], P<0.000 01) and in combination with other therapies(RR=1.18, 95%CI[1.14, 1.22], P<0.000 01) effectively improved the total effective rate.(2) In terms of symptoms/signs disappearance time, Chaihuang Granules effectively reduced the duration of fever(MD=-1.18, 95%CI[-1.78,-0.58], P=0.000 1), cough with sputum(MD=-1.82, 95%CI[-2.38,-1.25], P<0.000 01), cough(MD=-1.31, 95%CI[-1.89,-0.74], P<0.000 01), sore throat(MD=-1.57, 95%CI[-2.25,-0.89], P<0.000 01), and lung rales(MD=-1.49, 95%CI[-2.06,-0.92], P<0.000 01).(3) Regarding cytokine levels, Chaihuang Gra-nules effectively improved the levels of interleukin(IL)-2(MD=-0.94, 95%CI[-1.16,-0.72], P<0.000 01), IL-6(MD=-4.71, 95%CI[-6.39,-3.03], P<0.000 01), and tumor necrosis factor-α(TNF-α)(MD=-2.07, 95%CI[-2.43,-1.71], P<0.000 01).(4) In terms of cellular immune levels, Chaihuang Granules effectively improved the levels of CD3~+(MD=4.11, 95%CI[1.53, 6.69], P=0.002), CD4~+(MD=4.21, 95%CI[1.69, 6.73], P=0.001), CD8~+(MD=-2.65, 95%CI[-3.93,-1.37], P<0.000 1), and CD4~+/CD8~+(MD=0.25, 95%CI[0.14, 0.37], P<0.000 1).(5) In terms of humoral immune levels, Chaihuang Granules effectively improved the levels of IgA(MD=0.44, 95%CI[0.23, 0.64], P<0.000 1), IgM(MD=0.31, 95%CI[0.15, 0.46], P=0.000 1), and IgG(MD=2.02, 95%CI[1.60, 2.43], P<0.000 01). Trial sequential analysis showed that the cumulative Z-curve of the total effective rate crossed the boundary value, further confirming its clinical efficacy. The GRADE evidence quality evaluation showed that the evidence quality of the above outcome indicators was low or very low, and the recommendation strength was weak. Compared to conventional therapy alone, Chaihuang Granules can effectively improve the total effective rate of treatment, alle-viate symptoms and signs of upper respiratory tract infection in children, improve inflammatory conditions, enhance immune function, and reduce the recurrence rate. Due to the limited quality of the included studies, high-quality RCT is still needed to provide evidence support for the above conclusions.

Meta-analysis and trial sequential analysis of Compound Qinlan Oral Liquid in treatment of acute upper respiratory tract infection / 中国中药杂志

This study aims to evaluate the efficacy and safety of Compound Qinlan Oral Liquid in the treatment of acute upper respiratory tract infection. Computer-based online searching of CNKI, VIP, SinoMed, Wanfang, ChiCTR, ClinicalTrials.gov, Cochrane Library, PubMed, EMbase, and Web of Science was performed to retrieve the randomized controlled trial(RCT) regarding Compound Qinlan Oral Liquid in the treatment of acute upper respiratory tract infection. In addition, manual searching of gray literature was conducted. After two evaluators independently selected articles, extracted data, and evaluated the quality of methodology included in the studies, Meta-analysis was carried out in RevMan 5.4 and trial sequential analysis(TSA) in TSA 0.9.5.10 Beta. GRADE profiler 3.6.1 was employed to evaluate the evidence quality. A total of 21 RCTs were included in this study, involving 2 651 patients(1 330 patients in the observation group and 1 321 patients in the control group). Meta-analysis showed that compared with conventional western medicine alone, Compound Qinlan Oral liquid improved the total response rate(RR=1.15, 95%CI[1.12, 1.19], P<0.000 01) without increasing the incidence of adverse reactions(RR=0.77, 95%CI[0.47, 1.25], P=0.16). The results of subgroup analysis are described as follows:(1) Compared with conventional western medicine alone, Compound Qinlan Oral Liquid improved the total response rate(RR=1.10, 95%CI[1.05, 1.14], P<0.000 01) and shortened the time to symptom relief(SMD=-0.76, 95%CI[-1.02,-0.51], P<0.000 01). There was no significant difference in the incidence of adverse reactions between the two groups(RR=1.16, 95%CI[0.54, 2.47], P=0.71).(2) Compared with conventional western medicine alone, Compound Qinlan Oral Liquid + conventional western medicine improved the total response rate(RR=1.20, 95%CI[1.15, 1.25], P<0.000 01), decreased traditional Chinese medicine(TCM) syndrome scores(MD=-0.58, 95%CI[-0.75,-0.41], P<0.000 01), shortened the time to symptom relief(SMD=-2.44, 95%CI[-3.09,-1.80], P<0.000 01) and physical sign improvement(MD=-2.57, 95%CI[-4.11,-1.04], P=0.001), lowered the serum levels of inflammatory cytokines(SMD=-2.16, 95%CI[-2.61,-1.70], P<0.000 01), improved respiratory function indicators(SMD=1.48, 95%CI[1.00, 1.96], P<0.000 01), and enhanced the humoral immunity(MD=0.94, 95%CI[0.69, 1.18], P<0.000 01). There was no significant difference in the incidence of adverse reactions between the two groups(RR=0.57, 95%CI[0.29, 1.09], P=0.09). TSA showed that the cumulative Z curve of total response rate crossed the traditional threshold and TSA threshold, further confirming the clinical efficacy of Compound Qinlan Oral Liquid. The GRADE graded the evidence of the above outcome indicators as low or extremely low, and yielded weak recommendation. Compared with conventional western medicine alone, Compound Qinlan Oral Liquid can improve the total effective rate and reduce the time to symptom relief. The combination of Compound Qinlan Oral Liquid and conventional western medicine can improve the total response rate, mitigate the symptoms and improve the physical signs, reduce inflammation, and improve respiratory function and immunity of the patients with acute upper respiratory tract infection. In view of the limited number and quality of the included studies, the above conclusions still require high-quality RCT to provide evidence support.

Effects of Aconitum coreanum polysaccharides on TGF-β induced endothelial-mesenchymal transition and its mechanisms / 中国药理学通报

Aim To investigate the protective effect of Aconitum coreanum polysaccharides ( ACP) on TGF-β induced endothelial-mesenchymal transition ( EndMT) and its underlying mechanism. Methods HUVECs EndMT model was reproduced by 10 μg·L-1 TGF-β cultured for 72 h in vitro. The effect of ACP(50, 100, 200 mg·L-1) at various concentrations on TGF-β in-duced EndMT of HUVECs was explored. CCK8 assay was employed to detect the cell viability of HUVECs stimulated by ACP. Scratch test was used to analyze the migration ability of cells. The expressions of CD31, α-SMA, Smad2/3, p-Smad2/3 were deter-mined by Western blot. Results CCK-8 assay showed that various concentrations of ACP could promote HU-VECs viability obviously by 10 μg·L-1 TGF-β cul- tured for 36 h. Scratch test showed that TGF-β en-hanced transfer ability of HUVECs significantly, while this case was apparently reversed under the costimula-tion of TGF-β and ACP. Western blot showed that CD31 expression decreased significantly, and the ex-pressions of α-SMA, p-Smad2, p-Smad3 increased significantly, and this phenomenon could be regressed with ACP. Conclusion ACP can inhibit EndMT in HUVECs induced by TGF-β clearly, and the mecha-nism is related to down-regulation of p-Smad2, p-Smad3.

Specific sIgA antibodies against swine influenza virus H3N2 infection / 中国人兽共患病学报

To study different breed pigs reply the swine flu virus infections,specific antibody of sIgA secretion regularity of respiratory tract and the differences of sIgA antibody according to different antigen proteins were detected.A/swine/Nanjing/ 51/2010(H3N2) was intranasally infected pigs (1 × 107 TCID50/mL and 2 mL/pig),and then the nasal swab samples were collected at different time points within 21 days after infection.M1,NS1 and PB1 recombinant protein,respectively,were used to establish indirect ELISA method for detecting specific antibody of sIgA,and to analyze its secretion regularity.Results displayed that there was no significant difference among three kinds of recombinant protein in the whole test,characterizing by specificity sIgA antibody levels rising rapidly after 5 infection days and reaching peak at day 14,then began to decline.Among different varieties of pigs,sIgA antibody production of PB1 protein in Obama group was significantly higher than that in binary pigs at 14th and 21st day (P＜0.05).It had no significant difference between M1 group and the NS1 group (P＞0.05).This experiment preliminary explores the secretion regularity of specificity sIgA antibody after infected swine flu virus,which laid a foundation for further study of SIV mucosal antibody diagnostic reagents.

Analysis of Intervention Effect on Prophylactic Antibiotics Use in Type Ⅰ Incision in One Hospital / 中国药师

Objective:To analyze the effects of rational intervention on prophylactic antibiotics use in typeⅠIncision. Methods:A retrospective investigation method was used. Totally 170 cases with typeⅠIncision undergone in the hospital from April to Decem-ber 2013 were selected as the non-intervention group, and 173 cases with typeⅠIncision undergone in the hospital from April to De-cember 2014 were selected as the intervention group. The prophylactic antibiotics use, medication time, drug selection and duration of drug treatment were analyzed. Results:After intervention, the use rate of antibiotics was decreased from 67. 65% to 26. 59%, the ir-rational rate of use time was decreased from 15. 88% to 4. 63%, the irrational rate of treatment course was decreased from 30. 58% to 8. 09%, and the combination rate was declined from 10. 59% to 5. 20%. Conclusion:The rational prophylactic use of antimicrobial in type Ⅰ incision is improved through the intervention, and further efforts are still needed to improve the reasonable use.

Expression and significance of tumor drug resistance related proteins and beta-catenin in esophageal squamous cell carcinoma / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>As chemotherapy is generally used in the clinical treatment of cancer, the problem of multidrug resistance (MDR) of tumors against the chemotherapeutic agents becomes more and more serious. It has been the major cause for the failure of the chemotherapy. We detected the expressions of beta-catenin and tumor drug resistance related proteins, MRP2, P-gp, and Bcl-2, in esophageal squamous cell carcinoma (ESCC) to explore their function and correlation in the occurrence and development of MDR in ESCC.</p><p><b>METHODS</b>We used the tissue microarray technique, immunohistochemistry, and image analysis methods to detect the expressions of MRP2, P-gp, beta-catenin, and Bcl-2 proteins and analyze their relationships with clinical data in a ESCC tissue microarray consisting of 582 specimens of ESCC, 294 specimens of normal mucosa, 92 specimens of basal cell hyperplasia, and 87 specimens of dysplasia adjacent to cancer tissue.</p><p><b>RESULTS</b>The integral optical density (IOD) of MRP2 and Bcl-2, which was 195.7 +/- 175.9 x 10(3)) and 90.5 +/- 112.5 x 10(3)), respectively, was significantly higher in ESCC than in normal mucosa, which was 104.8 +/- 86.1 x103) and 25.2 +/- 46.6 x 10(3)), respectively (P < 0.01). The IOD of P-gp and beta-catenin, which was 57.7 +/- 75.5 x 10(3)) and 32.0 +/- 47.0 x 10(3)) respectively, was significantly lower in ESCC than in normal mucosa, which was 114.8 +/- 106.6 x 10(3)) and 46.1 +/- 35.7 x 10(3)), respectively (P < 0.01). According to the following order, well differentiated moderately differentiated poorly differentiated, the IOD of MRP2 increased in ESCC (P < 0.01). The IOD of beta-catenin was higher in poorly differentiated ESCC than in well or moderately differentiated ESCC (P < 0.01). The IOD of Bcl-2 was lower in well differentiated ESCC than in poorly and moderately differentiated ESCC (P < 0.01). The IOD of beta-catenin and Bcl-2 was higher in the ESCC of specimens with infiltration depths that were in membrane mucosa than those in the muscular layer or serous coat (P < 0.01). The IOD of Bcl-2 was significantly higher in cases with lymph node metastasis than in cases without (P < 0.01). Positive correlations which were respectively between the expressions of P-gp and MRP2, the expressions of P-gp and Bcl-2 were found (r = 0.288 and r = 0.253, respectively, P < 0.01).</p><p><b>CONCLUSIONS</b>MRP2, P-gp, and Bcl-2 may be taken as prognostic factors for MDR of ESCC. beta-catenin may play an important role in carcinogenesis and progression of ESCC.</p>

Effect of Xiaochaihu decoction and different herbal formulation of component on inhibiting H22 liver cancer in mice and enhancing immune function / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effects of Xiaochaihu decoction and different herbal formulation of componenton on inhibiting H22 mouse solid liver cancer and on the immune function, for approaching the dose-effect relationship and compatibility mechanism.</p><p><b>METHOD</b>Transplanted H22 liver the cancer to mice, filling on high, moderate, low dose of mice in the xiaochaihu decoction drug (27, 13.5, 6.75 g x kg(-1)), 5-FU intraperitoneal injection (20 mg x kg(-1)) as positive control. After being administered for 10 days, we determined the cancer weight, body weight, spleen index and thymus gland index for studying the dose-effect relationship. And we studied compatibility rule of Xiaochaihu decoction and the different herbal formulations of its components by using the same method. Mice in the Xiaochaihu decoction group (13.5 g x kg(-1)) and Ginseng-Chinese Date-Licorice Root apozem group (5 g x kg(-1)) were given the medicines for 10 days to observe the weight of tumor, body weight, spleen index, the NK cell activity, proliferation of the T lymphocytes and IL-2 level.</p><p><b>RESULT</b>Growth of H22 mouse liver cancer was markedly inhibited in high, moderate, low dose group; the inhibiting rates were 49.66%, 48.52%, 36.91%, respectively; the optimal administration dose was moderate. In the study of compatibility rule, only inhibiting rates of Ginseng-Chinese Date-Licorice Root apozem group and Xiaochaihu decoction group exceed 30%; the inhibiting rates were 41.55% and 43.65%. As compared with normal group, spleen index of three dose groups and the different herbal formulation groups was evidently increased (P < 0.01); thymus gland index of the different herbal formulation groups was significant decreased (P < 0.05); thymus gland index and body weight of 5-FU group were evidently decreased (P < 0.001). As compared the model group, the NK cell activity, proliferation of the T lymphocytes and IL-2 level of Xiaochaihu decoction were remarkably increased (P < 0.001); the NK cell activity and IL-2 level of Ginseng-Chinese Date-Licorice Root group were remarkably increased (P < 0.05); the NK cell activity and proliferation of the T lymphocytes and IL-2 level of 5-FU group were remarkable decreased (P < 0.01). As comparing with the normal control group ,body weight of 5-FU group was evidently decreased (P < 0.05).</p><p><b>CONCLUSION</b>Xiaochaihu decoction and Ginseng-Chinese Date-Licorice Root group can markedly inhibit the growth of H22 mouse solid liver cancer and improve the immune function of tumor-bearing mice. The inhibiting effect probably closely related with improving the tumor-bearing host immune function. There is a hint that compatibility Ginseng-Chinese Date-Licorice Root of strengthening body resistance is the core of inhibiting effect.</p>

Study on preparation of borneol-?-cyclodextrin inclusion complexes in Zhnyu Hewei Zhitong Capsules / 中华中医药杂志

Objective:To optimize the preparation of borneol-?-cyclodextrin inclusion complexes in Zhuyu Hewei Zhitong Capsules,and to establish the metered method of borneol in inclusion complexes.Methods:Orthogonal design was used and inclusion complex yield and inclusion rate of berneol were used as evaluation index to optimize the best preparation craft. The chromatographic conditions were:PEG-20M with 10%,3m??3mm of application concentration was used as stationary liquid phase,FID detector was used at 140℃,betula oil was used as internal standard substance and the sample was dissolved in Colonial spirit by ultrasounding.Results:The best preparation craft were:the weigh ratio of borneol and?-CD was 1:6,the ratio of?-CD and moisture addition was 1:10,reaction time was 30min,the temperature was 30℃.A good linearity was obtained when the concentration of borneol was between 0.7146-4.2878mg/ml(r=0.9999),the average recovery was 101.97%,RSD% was 1.77%.Conclusion:Three batches of tests indicated that the preparation of borneol inclusion complexes was stable and feasible. This assay method of borneol was accurate,sensitive and reproducible.