A quantitative nuclear magnetic resonance spectroscopic ( qNMR) method was established for the simultaneous determination of resveratrol and polydatin in Polygonum Cuspidatum traditional Chinese herb cuts and granule. The 2_step ultrasonic extraction method using 80% alcohol and acetone was used for fully extracting these two components in samples before qNMR determination. The qNMR experimental conditions were investigated and deuterated dimethyl sulphoxide_deuterium oxide (10∶1, V/V) was selected as solvent, the pulse delay time was 5 s, the scan number was 32, 2,3,5_triiodobenzoate was used as internal standard which was calibrated with primary standard substance of potassium hydrogen phthalate. The 1 H_NMR peaks atδ6. 388-6. 391 ( d, 2H) of resveratrol and δ 6. 322-6. 330 ( t, 1H) of polydatin were chosen as the quantitative peaks. Method validation was performed in terms of precision ( RSD<0. 6%), linearity (correlative constants R2>0. 999), limit of detection (0. 23 g/L for resveratrol and 0. 24 g/L for polydatin) and limit of quantitation ( resveratrol 0. 69 g/L, polydatin 1. 57 g/L), recovery ( resveratrol 97. 7% -103 . 5%, RSD=2 . 4%, polydatin 94 . 5%-99 . 2%, RSD=1 . 6%, including the sample extraction and preparaton process) . The results showed the reliability of qNMR for traditional Chinese medicine assay. The resveratrol and polydatin in Polygonum Cuspidatum real cuts and granule samples were experimental determined as 3. 57-5. 69 mg/g and 12. 73-24. 07 mg/g, respectively.

OBJECTIVE: To discuss the significance of Entecavir(ETV) in the treatment of chronic serious hepatitis B(HBV).METHODS: 54 patients with chronic serious hepatitis B were assigned to receive combined therapy in which antiviral drugs such as interferon and(or) nucleoside(acid) analogues antiviral drugs were excluded(control group,n=26) or combined therapy in combination with entecavir(0.5 mg?d-1) qd(treatment group,n=28).The course of treatment in both groups were 6 weeks.The hepatic function,HBV markers and HBV-DNA quantitation were deteted every two weeks.The improvement rate of patients after the completion of treatment were recorded.RESULTS: In the follow-up of six weeks,serum HBV-DNA and total bilirubin levels decreased markedly,and significant difference was noted between compared with the control group;ALT,AST,ALB and PT decreased in both groups,but the differnces between the two groups were not significant;there was no signficnant differnce in improvement rate between the treatment group and the control group(89.3% versus 84.6%).CONCLUSION: Entecavir can rapidly lower serum HBV-DNA level,downregulate bilirubin level,improve liver function,improve patients prognosis in patients with hepatitis B,thus it can be used to treat serious hepatitis B.However,used in short term,the survival rate of patients with severe hepatitis B can hardly be improved.

The method of quantitative nuclear magnetic resonance spectroscopy (qNMR) for determination of epigoitrin in Radix Isatidis was established based on solid phase extraction (SPE).The twice ultrasonic extraction method using pure water was used for fully extracting epigoitrin in sample, and then the extraction was enriched and concentrated by poly-Sery MCX SPE cartridge.The effect of sample pretreatment and qNMR experimental conditions was investigated.The qNMR experiment conditions were selected using DMSO as solvent, calibrated 2,3,5-triiodobenzoate as internal standard, and P1(pulse width)=14.1 μs, d1(pulse delay time)=5 s, NS(number of scan)=256.The .1H-NMR peaks of δ 5.365-5.399 (H-7b, d, 1H) of epigoitrin were chosen as the quantitative peaks.Method validation was performed including precision (intra-day precision RSD was 0.5%, and the inter-day precision was 0.8%), linearity (correlation coefficient r>0.9991), LOD (0.05 mg/g, standard curve method) and LOQ (0.19 mg/g, S/N≥150).The recoveries of the SPE-qNMR were 97.4%-101.7%.The result showed that the method was stable, accurate and reliable.With this method the epigoitrin in a real Radix Isatidis was determined to be <0.19-1.26 mg/g.SPE combining with qNMR could extend the application field of qNMR, especially in the detection of low-content component in complex samples.

BACKGROUND:Numerous clinical and basic studies have shown that electroacupuncture can improve the function of neurogenic bladder after suprasacral spinal cord injury. OBJECTIVE:To observe the effects of electroacupuncture on bladder function and connective tissue growth factor expression in rats with suprasacral spinal cord injury. METHODS:Forty-eight female Sprague-Dawley rats were randomly divided into four groups(n=12 per group):the blank group did not receive any treatment;the sham-operated group only exposed the T8 subvertebral spinal cord;in the model group established,a T8 subvertebral spinal cord transection injury model was established;in the electroacupuncture group,the T8 subvertebral spinal cord transection injury model was established,and electroacupuncture intervention at Ciliao(BL32),Zhongji(RN03)and Sanyinjiao(SP06)was given at 19 days after modeling,20 minutes once a day,for 10 continuous days.After the intervention,the relevant indicators were detected. RESULTS AND CONCLUSION:Urodynamics:Compared with the blank group,the leakage point pressure,maximum bladder capacity and maximum bladder pressure of rats in the model group increased(P<0.05).Compared with the model group,the leakage point pressure,maximum bladder capacity and maximum bladder pressure of rats in the electroacupuncture group decreased(P<0.05).Hematoxylin-eosin staining:Compared with the blank group,the bladder epithelial cells in the model group were arranged in a disordered manner,the lamina propria was destroyed,the detrusor muscle bundles were hypertrophied,the muscle fibers were arranged in a disordered manner,and the tissue edema was obvious.Compared with the model group,the bladder epithelial cells in the electroacupuncture group were arranged in a regular and orderly manner,and the degree of bladder fibrosis and tissue edema was relatively reduced.Masson staining:The degree of bladder detrusor muscle fibrosis was severe in the model group and it was lighter in the electroacupuncture group than in the model group.Transmission electron microscopy:Mitochondria in the bladder tissue in the model group were swollen and vacuolated,the morphology of the detrusor muscle was twisted and distorted,and the muscle gap was widened.Compared with the model group,mitochondria in the electroacupuncture group had a slightly clearer contour and were less vacuolated,and the muscle gap was narrowed.Western blot detection:The protein expression of connective tissue growth factor in the detrusor muscle of the bladder was elevated in the model group compared with the blank group(P<0.05).Compared with the model group,the protein expression of connective tissue growth factor in the bladder detrusor muscle was decreased in the electroacupuncture group(P<0.05).To conclude,electroacupuncture at Ciliao(BL32),Zhongji(RN03)and Sanyinjiao(SP06)acupoints can improve the morphology,structure and function of the bladder in rats with suprasacral spinal cord injury,and the mechanism of action may be related to the down-regulation of connective tissue growth factor protein expression in the detrusor muscle.