OBJECTIVE:To optimize the content determination conditions of total alkaloids from Zhuang medicine Munronia delavayi,and to determine the content of total alkaloids in M. delavayi from different producing areas. METHODS:With the con-tent of total alkaloids as index,using solvent amount,ultrasonic time,ultrasonic extraction times and pH value of buffer as fac-tors,the content determination conditions of total alkaloids from Zhuang medicine M. delavayi were optimized by orthogonal test. Optimized content determination conditions were adopted to determine the content of total alkaloids in M. delavayi from 18 produc-ing areas in different harvest time. RESULTS:The optimum content determination conditions were as follows as the amount of sol-vent(CHCl3)20 ml,ultrasonic processing for 3 times,lasting for 15 min each time,pH value of buffer 4.5. The contents of total alkaloids in M. delavayi from 18 producing areas were between 0.6-11.98 mg/g,showing great difference. M. delavayi from Long-lin county and Tianlin county harvested in Oct. had the lowest content of total alkaloids. CONCLUSIONS:Optimized content deter-mination condition can be used for the content determination of total alkaloids in Zhuang medicine M. delavayi and quality control of it. The content determination of total alkaloids in M. delavayi is related to producing area and harvest time.

Objective Benign prostatic hyperplasia ( BPH) is one of common diseases in aged males , and searching for new therapeutic drugs to BPH has been a research hotspot in recent years .This article was to study the inhibitory effect of eupatorium ja-ponicum thunb and foeniculum vulgare extract ( EFE) on benign prostatic hyperplasia in rats and its possible mechanism . Methods 48 male rats were randomly divided into 6 groups:normal control group without any treatment , model group of BPH treated with subcu-taneous injection of testosterone propionate , positive control group of BPH treated with dutasteride , high, middle and low dosage groups according to different EFE dosage (156 mg/kg, 234 mg/kg and 312 mg/kg).45 days after the treatment, the rats were sacrificed for measurement of the prostate glandular wet weight , the index of prostate gland ( PI ) , the morphological changes of prostate gland by light microscopy and the content of sex hormone . Results The prostate wet weight and PI decreased after EFE treatment for 45 days compared with the BPH model group(P<0.01 ).The hyperplastic glandular epithelium papilla waned and even disappeared in three EFE groups under the light microscope , and the epithelial cells became cubical or flat .High dosage EFE group (312 mg/kg) has simi-lar efficacy to dutasteride group .EFE significantly reduced serum testosterone content , dihydrotestosterone content and T/E2 ratio( P<0.05 ). Conclusion EFE can significantly inhibit prostatic hyperplasia in rats , and its mechanism is related to the decrease of the contents of serum testosterone and dihydrotestosterone as well as T/E2 ratio.

OBJECTIVE:To establish a method for the simultaneous determination of scutellarin and linarin in Scoparia dulcis. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase of acetonitrile-0.05% phosphoric acid (gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 330 nm,column temperature was 30 ℃,and the in-jection volume was 10 μl. RESULTS:The linear range was 0.015 8-0.316 8 mg/ml for scutellarin(r=0.999 9)and 0.000 5-0.010 4 mg/ml for linarin (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 99.56%-102.38%(RSD=1.07%,n=6)and 95.14%-98.29%(RSD=1.24%,n=6),respectively. CONCLUSIONS:The method is simple with good stability and reprodicibility,and can be used for the simultaneous determination of scutellarin and linarin in S. du-lcis.

OBJECTIVE: To investigate the effects of Xiehuo Bushen Decoction (XHBSD), a compound Chinese herbal medicine, on the survival and differentiation of transplanted neural stem cells (NSCs) in brains of rats with intracerebral hemorrhage, and to explore the mechanism of Xiehuo Bushen formula in promoting the survival of transplanted NSCs. METHODS: NSCs separated from hippocampuses of neonatal SD rats were cultured. Sixty-five panel reactive antibody (PRA) positive SD rats were selected by lymphocytotoxicity methods. The PRA positive rats were made into intracerebral hemorrhagic model and divided into three groups: cerebral hemorrhage group (n=15), NSCs transplanted group (n=25) and XHBSD group (n=25). XHBSD was orally administered after 5-bromodeoxyuridine (BrdU)-marked NSCs were transplanted in brains of rats with intracerebral hemorrhage in the XHBSD group. Rats in the other two groups were administered distilled water. The expressions of interferon gamma (IFN-gamma) and interleukin-4 (IL-4) mRNAs were measured by reverse transcription polymerase chain reaction (RT-PCR); the numbers of BrdU and 200 kD neurofilament (NF200) positive cells were detected by double-labeling immunofluorescence method. RESULTS: The expression of IFN-gamma mRNA was down-regulated significantly in the XHBSD group, but the expression of IL-4 mRNA was up-regulated significantly (P<0.05). The numbers of BrdU and NF200 positive cells were also increased remarkably in the XHBSD group. CONCLUSION: XHBSD can promote the survival and differentiation of transplanted NSCs, which may be related to inducing the expression of IL-4 mRNA and inhibiting the expression of IFN-gamma mRNA.

AIM: To observe the expression of fractalkine, and its receptor, CX3CR1, in renal tissues of patients with diffuse proliferative lupus glomerulonephritis (WHO class IV), minimal glomerular abnormalities, and normal kidney. Meanwhile, the correlation among the expression of fractalkine, CX3CR1 and CD68-positive macrophages was investigated, and the role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was discussed. METHODS: The expressions of fractalkine, CX3CR1 and CD68 were detected immunohistochemically in kidney tissue sections obtained from twenty-one patients with WHO class IV lupus nephritis, eighteen cases with minimal glomerular abnormalities, and eight normal kidneys which were no abnormality under light microscope. RESULTS: (1) Fractalkine was generally indistinguishable in tissue sections from normal kidney and minimal glomerular abnormalities. CX3CR1-positive cells and CD68-positive macrophages were sparsely detected in the glomeruli and in the cortical interstitium. (2) There were considerable CX3CR1-positive cells and macrophages in both the glomeruli and the interstitium in sections from class IV lupus nephritis. The number of CX3CR1-positive cells significantly correlated with the number of macrophages in the glomeruli and in the interstitium respectively (r=0.956, P

Objective To observe the analgesic and anti-inflammatory effects of the water extract of Glycosmis citrifolia (Willd.) Lindl.on mice and explore the mechanism. Methods The analgesic and anti-inflammatory effects were evaluated by 0.7% acetic acid-induced writhing test,the hot plate test,tests of dimethylbenzene-induced ear swelling,1% carrageenan-induced paw edema,determination of PGE2 in inflammatory feet,0.6% acetic acid-induced increase in peritoneal capillary permeability and cotton ball granuloma. Results The water extract of Glycosmis citrifolia (Willd.) Lindl.at low,medium and high doses can reduce the acetic acid-induced writhing times (P<0.01 or P<0.05),increase the pain threshold of mice (P<0.01 or P<0.05), inhibit dimethylbenzene-induced ear swelling (P<0.01 or P<0.05),1% carrageenan-induced paw edema (P<0.01 or P<0.05) and PGE2 production (P<0.01),0.6% acetic acid-induced increase of peritoneal capillary permeability (P<0.05),and the de-velopment of cotton ball granuloma (P<0.01 or P<0.05). Conclusion The water extract of Glycosmis citrifolia (Willd.) Lindl.shows analgesic and anti-inflammatory effects on mice.

Objective To investigate the effects of acyl-CoA synthetase 5 (ACS5) silencing by siRNA on expression and proliferation of colon carcinoma cell lines.Methods The expression of ACS5 in 30 case colon carcinoma and adjacent tissues were analyzed by immunohistochemical staining.The siRNA of ACS5 with Lipofectamine2000TM was transfected into colon carcinoma cell lines (HT-29 and SW480).The expression of ACS5 in colon carcinoma cell lines (HT-29 and SW480) was detected by real-time reverse transcription polymerase chain reaction.Proliferation of colon carcinoma cell lines was analyzed by 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS).Results The expression of ACS5 in colon cancer was significantly higher than in adjacent tissues by immunohistochemical staining.The mRNA of ACS5 in siRNA-ACS5 group (0.18 ± 0.03) was significantly lower than in NC siRNA group (2.55 ± 0.31) and blank control group (2.48 ± 0.12) in HT-29 colon cancer lines,and the inhibition ratio was 92.96％ (F =146.9,P ＜0.01).The mRNA of ACS5 in siRNA-ACS5 group (0.14 ± 0.01) was significantly lower than in NC siRNA group (1.21 ± 0.05) and blank control group (1 ± 0.03) in SW480 colon cancer lines,and the inhibition ratio was 88.5％ (F =826.5.9,P ＜ 0.01).Proliferation of HT-29 and SW480 colon cancer line in siRNA-ACS5 group was slower on 72 h and 96 h than in NC siRNA group and blank control group (P ＜ 0.05).Conclusions Expression of ACS5 is elevated in colon cancer tissues.siRNA interference of colon cancer line downregulated ACS5 expression and inhibited the proliferation of the colon cancer cells.

The lack of effective in vitro infection model for hepatitis E virus (HEV) has greatly hindered the quantitative analysis of neutralizing titers of anti-HEV antibodies and human sera, thus impeding further studies of HEV-stimulated antibody responses and the immunological mechanisms. In order to improve this situation, the infection of HepG2 cells that are inefficient for HEV replication was continuously monitored until the viral load reached the limit of detection on day 13, the results of which confirmed the feasibility of using this cell line to establish the infection model. Then, neutralization assays of five anti-HEV murine monoclonal antibodies and serum samples collected from four HEV vaccine recipients (collected before and after vaccination) were performed by 96 multi-channel parallel infections, nucleic acid extraction, and qPCR. The results showed that the cell model can be applied for quantitative evaluation of the neutralizing capacity of different antibodies and antiserum samples from HEV vaccine recipients. In this study, we have successfully established a high-throughput in vitro HEV replication model, which will prove to be useful for the evaluation of HEV vaccines and studies of HEV epitopes.
Animals ; Antibodies, Viral ; analysis ; immunology ; Hepatitis Antibodies ; analysis ; immunology ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; chemistry ; immunology ; physiology ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Neutralization Tests ; methods ; Virus Replication

Objective To investigate the incidences, risk factors, genotypes and epidemiology of community-acquired blood stream infection caused by extended spectrum β-lactamases (ESBLs)-producing Escherichia coli and Klebsiella pneumonia strains and to analyze the sensitivity of those ESBLs producing strains to commonly used antibiotics. Methods Forty-two patients who were diagnosed with community-ac-quired blood stream infection caused by Escherichia coli or Klebsiella pneumonia strains in Sichuan Provincial People′s Hospital were recruited in this study. Disc diffusion method was used for the phenotypic confirmato-ry test of ESBLs. Agar dilution method was performed to measure the antimicrobial susceptibility of the ESBLs-producing strains to 13 clinically commonly used antibiotics. Genotypes of the ESBLs-producing strains were identified by polymerase chain reaction (PCR). Multilocus sequence typing (MLST) was used to analyze the epidemiology of ESBLs-producing strains. Logistic regression analysis was performed to analyze the risk factors for community-acquired blood stream infection. Results The ESBLs-producing Escherichia coli strains accounted for 56. 3% (18 / 32) and the ESBLs-producing Klebsiella pneumoniae strains accounted for 20% (2 / 10). All of the 20 ESBLs-producing strains were sensitive to imipenem, meropenem, ertapen-em, nitrofurantoin and moxalactam. The ESBLs-producing strains sensitive to amikacin, piperacillin-tazobactam and fosfomycin accounted for 95% , 90% and 85% , respectively. Drug resistance rates of the 20 strains to cefotaxime, levofloxacin, ciprofloxacin and cefepime were relatively high accounting for 100% , 80% , 80% and 75% , respectively. Among the 20 ESBLs-producing strains, 7 strains only carried the CTM gene, while the other 13 strains were all positive for two genotypes of ESBLs, mainly identified as TEM+CTM-M-14 and TEM+CTM-15 genotypes. The 18 Escherichia coli strains were classified into 10 ST types, most of which were ST131 type, followed by ST10 and ST38 types. This study indicated that malignant tumor might be a possible risk factor. Conclusion The prevalence of community-acquired blood stream infection caused by ESBLs-producing Escherichia coli strains was becoming increasingly serious. Malignant tumor might be the risk factor associated with the producing of ESBLs in Escherichia coli and Klebsiella pneumonia strains. TEM+CTX-M-14 was the predominant genotype of ESBLs-producing strains and the prevalent clone was ST131 type. Carbapenems and enzyme inhibitor compounds were ideal drugs for the treatment of commu-nity-acquired blood stream infection caused by ESBLs-producing Escherichia coli and Klebsiella pneumonia strains. This study was limited by the small sample size. Therefore, it is necessary to conduct further resear-ches based on a large number of samples.

Objective To evaluate the clinical effects of color Doppler ultrasound in perforating vein closure by foam sclerotherapy for the treatment of venous leg ulcer.Methods Choosing the patients with venous ulcers (classified as C6) that underwent superficial and perforating veins closure by ultrasound-guided foam sclerotherapy in the Department of Vascular Surgery,First Affiliated Hospital of Chongqing Medical University,during December 2014 to August 2016.All patients were followed up by color Doppler scanning for veins closure,and the postoperative healing of skin ulcers were observed at the same time.Results A total of 114 patients with 119 limbs were enrolled,which were confirmed 1-3 perforating vein lesions by ultrasound.An average of 2.1 perforators were closed per leg (1-3 perforators).The average follow-up period was 11.3 months (1-21 months).The healing rate is 100％ and all the insufficient perforating veins were occlusion 1 month postoperative.Three cases of ulcer recurrence,and 8 cases of insufficient perforator recanalization were found.Serious complications did not appear.Conclusions Color doppler ultrasound can improve the safety and efficacy of perforating vein closure by foam sclerotherapy for the treatment of venous leg ulcer,and it has a high clinical value.