Objective To design and synthesize derivatives of wanpeinine A, the main steroidal alkaloid isolated from the plant Fritillaria shuchengensis, and further study on the structure-activity relationship of the steroidal alkaloid. Methods Acylation and alkylation were used to synthesize the derivatives and their structures were identified via NMR and MS.Results The acylation of wanpeinine A (1) produced 3β,6α-diacetylwanpeinine A (2), 3β,6α-dipropionylwanpeinine A (3), 3β,6α-dichloracetylwanpeinine A (4), 3β,6α-dibenzoylwanpeinine A (5), and 3β-methoxyacylwanpeinine A (6). The alkylation of wanpeinine A formed 3β,6α-dimethoxymethylwanpeinine A (7). Conclusion All compounds are new except for 3β,6α-diacetylwanpeinine A.

OBJECTIVE To explore the phenotype and genotype of Klebsiella and Escherichia coli producing extended-spectrum ?-lactamases(ESBLs).METHODS The producing ESBLs strains in 110 E.coli and 94 Klebsiella isolates were determined according to disk diffusion test,recommended by NCCLS and the drug-resistant genes such as TEM,SHV,CTX-M-1,CTX-M-2 and CTX-M-9 were detected by PCR and sequencing.RESULTS The producing ESBLs rates were 36.2%,42.7% and 39.7% in Klebsiella,E.coli,and in both two strains,respectively.The positive rate in detecting ESBLs by CTX and CTX/CLAV was higher(66.7%);the negative rates were 68.1% in E.coli,64.7% in Klebsiella when using CAZ alone and CAZ/CLAV.The rates of TEM,SHV,CTX-M-1,CTX-M-2 and CTX-M-9 were 54.3%,38.3%,21.0%,24.7% and 70.4%,respectively,in producing ESBLs isolations.The CTX-M genotype was predominant,(91.4%);the isolations(71.6%) contained more than two resistance genes.CONCLUSIONS More attention should be payed on ESBLs producing E.coli and K.pneumoniae strains.

One hundred and seventy six consecutive patients with acute ischemic stroke who received intravenous recombinant tissue plasminogen activator ( rt-PA ) in 4.5 hours from symptom onset during February 2009 to July 2013 were included in the study.Modified Rankin Scale was used to evaluate the recovery of neurological functions.Patients were divided into good ( 0 -1 ) or poor ( 2 -6 ) outcome groups according modified Rankin Scale score.Univariate analysis and multivariate logistic regression analysis were used to determine the differences of clinical data between the two groups.The age of patients with good outcome was significantly lower than that of poor outcome group [ ( 61.4 ±11.5 ) vs.( 69.0 ± 13.2) years,P =0.000].Compared to patients with poor outcomes, patients with good outcome group showed lower rate of diabetes [ 13%( 12/93 ) vs.29%( 24/83 ) , P =0.009 ] , lower blood glucose level [(5.05 ±0.97) vs.(5.83 ±1.72) mmol/L,P=0.020], higher uric acid level[(404.4 ±151.7) vs.(345.6 ±107.5) μmol/L,P=0.028],shorter onset to treatment time [(1.92 ±0.94) vs.(2.30 ±1.01) h, P=0.019],lower baseline National Institute of Health Stroke Scale score [(14.0 ±5.2) vs.(16.0 ± 6.2),P=0.025],lower systolic blood pressure level at 2 h[(140.8 ±18.3) vs.(149.0 ±18.9) mmHg (1 mmHg=0.133 kPa),P=0.005]and 24 h [(137.6 ±21.9) vs.(147.1 ±17.4) mmHg,P=0.009] after thrombolysis.Logistic regression analysis showed that uric acid levels were not related to hemorrhagic transformation independently (P =0.172,OR =0.965,95%CI:0.917 -1.016), but were related to outcome independently (P=0.047,OR=0.957,95%CI:0.916-0.999).

Objective To express the capsid proteins of WU polyomavirus(WUPyV) for research and find antigen for diagnostic value. Methods Coding sequences of capsid proteins of WU polyomavirus by PCR were cloned in prokaryotic expression vector PGEX-20T. Recombinant plasmids were transformed into E. coli BL21(DE3) and induced by IPTG for proteins expression. Recombinant proteins were identified by Western blot. Results SDS-PAGE proved that recombinant proteins showed three bands with molecular relative mass of 69×103, 63×103 and 56×103. The recombinant proteins were recognized by anti-GST McAb. The antigenicity was tested by Western blot using 16 WU polyomavirus positive and 70 negative sera. Conclusion Recombinant VP1, VP2 and VP3 expressed in E. coli can combine with WUPyV-Ab and have good antigenicity. They can be used for further research.

Objective To establish high performance liquid chromatography(HPLC)characteristic chromatograms of Xinqingduyin Granules(composed of Taraxaci Herba,Lonicerae Japonicae Flos,Chrysanthemi Indici Flos,etc.)and content determination of chicory acid and glycyrrhizic acid,and to optimize the preparation process of Xinqingduyin Granules.Methods Using the characteristic chromatograms of Xinqingduyin and the retention rate of chicory acid and glycyrrhizic acid as indexes,we carried out orthogonal experiment to optimize the extraction process of Xinqingduyin,and studied the concentration process.The molding process of Xinqingduyin Granules was conducted by screening the types and dosage of auxiliary materials,then three batches of pilot experiments were carried out.Results HPLC characteristic chromatograms of Xinqingshuyin Granules and the determination methods of chicory acid and glycyrrhizic acid were established.The optimal preparation technology was as follows:8 times amount of water was added,the drug was decocted for 3 times,with 1 hour per time.After the extract was concentrated under reduced pressure at 80℃,the appropriate amount of steviol glycoside and lactose was added into the extract and mixed.One-step granulation and packaging were adopted.The retention rates of chicoric acid and glycyrrhizic acid in the 3 batches of Xinqingduyin Granules,which were prepared on the pilot scale,were(54.56±1.63)%and(54.96±1.08)%,and the rate of finished product was(87.47±0.49)%,respectively.The quality is uniform,and the characteristic map of Xinqingduyin Granules showed high similarity with that of decoction prepared from the same batch of slices.Conclusion The optimized preparation technology is reasonable,feasible and reproducible.This preparation can be used to obtain the granule with similar materials of Xinqingduyin decoction.