Objective To develop and validate a simple,rapid,sensitive,and reproducible HPLC method for determination of hyperoside in plasma of dogs and for the subsequent pharmacokinetic (PK) study.Methods An accurate and reproducible HPLC-UV method was developed and validated for the determination of hyperoside in plasma of dogs,using kaempferol as internal standard.The plasma samples of dogs following ig administration of hyperoside were analyzed for the detection of quercetin after enzymatic hydrolysis treatment with combined β-glucuronidase and sulphatase.The analytes were separated on a Diamonsil C18 column (250 mm × 4.6 mm,5 μm).The mobile phase consisted of methanol-buffer solution (0.1mol/L NH4Ac + 0.3 mmol/L EDTA-Na2)-acetic acid (60:40:1) and was delivered at a flow rate of 1mL/min.The UV detector was set at 370 nm and the column temperature was maintained at 35 ℃.The sample injection volume was 20 μL.Data were collected and analyzed using the ANASTAR software.PK parameters were calculated with DAS software (2.0).Results Linear relationships were validated over the range of 0.01-1μg/mL for hyperoside (r =0.9997).The intra-and inter-day precision values for all samples were within 10.0％,and the accuracies of intra-and inter-day assays were within the range of 92.4％-102.4％.The validated method was successfully used to determine the hyperoside concentration in plasma of dogs for up to 12 h,after a single ig administration (25 mg/kg).The mean PK parameters for male and female dogs were as follows:Cmax (0.18 ± 0.05) and (0.16 ± 0.05) μg/mL,AUC0-∞ (0.79 ± 0.34) and (0.86 ± 0.27) μg/(mL·h),t1/2(ka) (0.89 ± 0.41) and (0.88 ± 0.28) h,respectively.Statistical analysis on the PK of hyperoside in male and female groups showed that sex had no significant impact on the PK of hyperoside (P ＞ 0.05).Conclusion The method is able and sufficient to be used in drug PK studies of hyperoside.

Objective To study the renal protective effects of superfine powder of Rhodiola rosea in diabetic nephropathy (DN)rats.Methods The DN rats model was established by one side nephrectomy and intraperitoneal injection of streptozotocin(STZ).Then rats were randomly divided into 6 groups:sham-operation group,model group,Rhodiola rosea common powder groups(3 g/kg·d-1),and superfine powder of low,medium and high dose groups (0.75、1.5、3.0 g/kg·d 1),10 rats in each group.Rats were given the relevant drugs respectively for 7 weeks.Finally the levels of fasting blood glucose (FBG),serum total cholesterol (STC),blood urea nitrogen (BUN),creatinine (Cr) and urinary protein (Upro) were detected.The kidney tissue was taken for histopathological observation.Results Compared with DN modelgroup [FBG,STC,BUN,Cr and Upro was (24.78±3.66)mmol/L,(4.07±1.32)mmol/L,(22.36±2.54)mmol/L,(86.78±7.47) μmol/L,(50.23±6.82)mg respectively],the rats in DN model group after treatment with common and superfine powder (medium and high dose groups) of Rhodiola rosea appeared different levels of improvement ofFBG [(18.67±2.74) mmol/L,(17.21 ±3.17)mmol/L,(15.48±2.46) mmol/L],STC [(3.33±0.87) mmol/L,(3.42 ± 0.74) mmol/L,(3.21 ±0.92)mmol/L],BUN [(15.43±2.04)mmol/L,(16.56±1.85)mmol/L,(12.44±1.36)mmol/L],Cr [(66.17 ± 4.43) μmol/L,(68.42 ± 5.06) μmol/L,(61.33±4.21)μmol/L] and Upro [(37.47±5.64) mg、(35.66 ± 4.72) mg,(27.37± 3.92) mg],P＜ 0.05.The pathologic changes of kidney tissue were improved,too.Conclusion Rhodiola rosea has a good renal protective effect in rats with DN.Furthermore,the superfine powder is more effective than the common powder in renal protective effects.

Aim To observe the anti-hepatitis B virus effect of total phenolics acid extracted from Oenanthe Javanica(OJTPA) with in vitro experiments.Methods HePG2 2.2.15 cells were treated with OJTPA,which were diluted into 500 mg?L-1,250 mg?L-1 and 125 mg?L-1 groups.The test set up positive control group(entecavir,3 mg?L-1) and normal control group.The cells of the 4 th,8 th and 4 th day ceasing treatment were collected.The content of HBV-DNA and cccDNA was determined with Quantitative Real-Time PCR method,and the inhibition rates were calculated.Results OJTPA could obviously inhibit HBV-DNA and cccDNA under the dosages used in the experiment,and showed a dose-dependent relationship.The inbhibitory rates reached the peak of 62.3% and 62.7% for HBV-DNA and cccDNA respectively,and a high inhibitory rate was retained within 3 days of OJTPA withdrawal.Conclusion OJTPA has significant inhibition on HBV-DNA and cccDNA and there is no significant rebound phenomenon.

This paper statistically analyzes sixty-two articles concerning the treatment of obesity with acupuncture, and finds that the common therapy embraces needling, moxibustion, electroacupuncture, ear acupuncture, auricular application, auricular needle-embedding, acupoint application, acupoint embedding and combined methods of acupuncture. The frequent auricular acupoints are Stomach (MA-IC), Endocrine (MA-IC 3), Er Shenmen (MA-TF 1), Lung (MA-IC 1), Spleen (MA-IC), Hunger Point (MA-T) and Sanjiao (MA-IC 4). The clinical total effective rate was 75.8%.

Objective To analyze the factors that influence joint function after plate fixation of unstable pelvic fractures.Methods A retrospective study was conducted to analyze the 109 patients with unstable pelvic fracture who had been treated with plate fixation from January 2007 to September 2014.They were 69 men and 40 women,23 to 72 years of age (average,46.3 years).By the Tile classification,15 cases were type B1,37 type B2,14 type B3,19 type C1,16 type C2,and 8 type C3.Their postoperative joint function was evaluated according to the Majeed criteria for functional evaluation.There were 86 cases in the good-to-excellent group (78.9％) and 23 in the fair-to-poor group (21.1％).Their data were analyzed in terms of gender,age,preoperative injury severity scale (ISS),body mass index (BMI),operation time,fracture type,reduction quality,postoperative complications,bone density,and postoperative weight bearing time.The influential factors were determined using Logistic regression analysis.Results All the 109 patients obtained a mean follow-up of 18.7 months (from 12 to 53 months).Preoperative ISS (P =0.000),fracture type (P =0.008),reduction quality (P =0.009) and postoperative complications (P =0.000) were identified as the factors influencing joint function after plate fixation of unstable pelvic fractures.Conclusion Preoperative assessment by ISS,a clear understanding of the pelvic fracture type,anatomical reduction of the fracture and efforts to reduce postoperative complications can effectively improve functional recovery of the joint after plate fixation of unstable pelvic fractures.

Objective To analyze the change of schistosomiasis endemic situation in Hubei Province, 2008. Methods The data of schistosomiasis control in the whole province, surveillance sites and standard examinations were collected and analyzed. Results Among all the 63 endemic counties (cities, districts), 21 reached the criteria of transmission interruption, 19 reached the criteria of transmission control, and 23 reached the criteria of infection control in 2008. Villages of Category Three, Category Four and Category Five accounted for 52.10% , 27.80% and 20.09% of all the endemic villages in the whole province, respectively. The infection rates of population and cattle were 1. 70% and 2. 20% , which decreased by 27. 04% and 34. 33% when comparing to those in 2007. The snail area was 77 363 hm~2, with a reduction of 315.70 hm~2 comparing to 2007. In the 74 provincial surveillance sites, the infection rate of population and cattle were 1. 32% and 1. 85% , respectively, with reduction rates of 32.99% and 68.38% comparing to 2007. The snail area was 2 833.98 hm~2 and the area with infected snails was 215.35 hm~2, the densities of living and infected snails were 0. 37 snails/0.1 m~2 and 0.000 3 snails/0. 1 m~2, respectively, and the infection rate of snails was 0.08%. Comparing to 2007, the snail area, densities of living and infected snails and the infection rate of snails decreased by 3.81 % , 21.28% , 40.00% and 27. 30% , respectively, while the area with infected snails increased by 4.52%. The results of standard examination suggested that all the 15 uncontrolled counties ( cities, districts) reached the criteria of infection control. Conclusions Though the endemic situation of schistosomiasis in Hubei Province has been controlled effectively, the endemic condition is still severe. Therefore, the comprehensive measures with an emphasis on infectious source control should be strengthened to consolidate the control result.

Objective To investigate the effects of artesunate (Art) on cell apoptosis, tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) expression induced by high glucose in rat renal tubular epithelial cells (NRK-52E). Methods NRK-52E cells were cultured and divided into normal control group, high glucose group, high glucose with different concen?trations of Art (10 mg/L, 20 mg/L and 30 mg/L) groups, and high glucose with Ena (5 mg/L) group. MTT assay was used to de?tect the cell proliferation. The apoptotic rate was evaluated by flow cytometry with AnnexinV-FITC/PI double stains. The pro?tein levels of TNF-αand IL-8 in the cell culture supernatant were determined using ELISA. Results High glucose inhibit?ed NRK-52E proliferation, induced its apoptosis, and the expressions of TNF-αand IL-8 in the supernatant. Application of Art obviously abolished the effects of high glucose, and the effects of Art were showed in dose-dependent manner. Conclu?sion Art can suppress high glucose-stimulated cell apoptosis, enhance TNF-αand IL-8 expression in NRK-52E cells. The anti-inflammatory action and immune regulation of Art could be a novel approach of treating diabetic nephropathy.

OBJECTIVE To investigate the effect and related mechanism of all￣trans retinoic acid (atRA) exposure on osteogenic differentiation of mouse embryonic palate masenchymal cells MEPM. METHODS MEPM were cultured in osteogenic medium (OM) with atRA 0.1 and 1.0 μmol??L-1 for 1, 3,5, 7 and 9 d. MTT assay was performed to measure the cell viability. The alkaline phosphatase (ALP) activity was measured by chemical colorimetry. The cells were stained using the Von￣Kossa technique to detect the formation of mineralization nodules after 21 d of culture. RT￣PCR was performed to determine expression Runx2, osteopontin, bone morphogenetic protein receptor ( Bmpr) 1b, Bmpr2 and Smad5 mRNA. RESULTS The result of MTT on 9 d showed that, compared with normal control group, the cell viability of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups decreased significantly(P<0.01). Compared with normal control group, ALP activity of OM group increased significantly(P<0.05), while the ALP activity of OM+atRA 0.1 and 1.0 μmol??L-1 groups was lower than OM group(P<0.05). On 21 d, the Von￣Kossa stai￣ning results showed that the percentage of mineralization nodules formation of OM+atRA 1.0 μmol??L-1 group was (3.65±1.24)%, which was significantly lower than that of OM group(10.33±2.29)%(P<0. 05). On 9 d, the relative Run expression of OM group was the highest one in the four groups, while at￣RA 1.0 μmol??L-1 treatment negatively regulated 20% in comparsion with OM group(P<0.05). Compared with normal control group, the mRNA expression of osteopontin of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups increased significantly(P<0.05); BDNF mRNA expression of OM group was 2.6￣fold to normal control group, while that of OM+atRA 1.0 μmol??L-1 group was 33% to OM group(P<0.05) . The level of Smad5 mRNA of OM+atRA 1.0 μmol??L-1 group was significantly lower than that of OM group(P<0.05). CONCLUSION atRA Might inhibit osteogenic differentiation of MEPM by down￣regulated the expression of Bmpr1b.

The inundation of Cordyceps sinensis counterfeits in the market makes it difficult to identify. In this study, 21 batches of wild C. sinensis from 3 different regions, 30 batches of naturally cultured C. sinensis and 4 kinds of counterfeits extracted by methanol and water were analyzed using NMR technology. 9 characteristic peaks were defined as quantitative criterion after comparison, and NMR fingerprints of C. sinensis were established. According to the result it is highly similar between naturally cultured C. sinensis and wild ones by comparing their NMR fingerprints. However, NMR spectra of four kinds of adulterants showed differences with C. sinensis. The result also showed that NMR fingerprint of C. sinensis are highly characteristic and specific. The NMR characteristic fingerprint of wild C. sinensis was consistent with the naturally cultured C. sinensis, and it indicated that the chemical constituents of wild C. sinensis and naturally cultured C. sinensis are nearly the same.

Objective To assess the schistosomiasis endemic situation in the national surveillance sites in Hubei Province, so as to provide the evidence for the prevention and control of schistosomiasis. Methods According to the national surveillance protocol,a longitudinal surveillance of endemic situation of schistosomiasis was carried out in 16 national surveillance sites from 2005 to 2010. Results In general,the positive rates of IHA,Kato-Katz technique and infection rates of Schistosoma japonicum decreased from 15.67%,10.93%and 1.71%in 2005 to 10.48%,8.54%and 0.90%in 2010,and declined by 33.12%,22.70%and 47.95%,respectively. The infection rates of S. japonicum of the male were higher than that of the female,and the peak infec-tion rates were in the groups aged above 30 years. The endemic situation of fishermen and farmers were relatively serious. The in-fection rates of S. japonicum in cattle decreased from 11.69%in 2005 to 1.41%in 2010,and declined by 88.01%(χ2 =298.79, P<0.001). The areas with infected Oncomelania hupensis snails,the densities of living snails,the rates of infected snails and the densities of infected snails decreased by 90.88%,61.66%,80.00%and 92.00%,respectively. Conclusion The schistosomiasis endemic situation in the national surveillance sites in Hubei Province mitigates in 6 years,but the prevention is still a very daunt-ing task.