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OBJECTIVE:To isolate and purify three principal degraded impurities of mitiglinide calcium (impurity A,B,C) and identify their structures,establish HPLC method for content determination of impurity A,B,C. METHODS:Mitiglinide calci-um was used as raw material and reacted with acid;3 impurities were then separated by HPLC and their structures were elucidated by IR,MS,1H NMR,13C NMR,LC-ESI-MS and ORD. 3 impurities of 3 batches of mitiglinide calcium were determined,and the determination was performed on Agilent Extend-C18 column with mobile phase consisted of 0.01 mol/L sodium acetate solution-ace-tonitrile-triethylamine(60:40:0.1,pH=3.0)at the flow rate of 1.0 ml/min. The detection wavelength was set at 210 nm and sam-ple size was 20 μl. The response tests of 3 impurities and mitiglinide calcium were conducted. RESULTS:After treated with acid, impurity A,B,C had been obtained,and their purity were 99.05%,98.87%,99.98%,respectively after isolation and purifica-tion;after identifying the structure, 3 impurities were S-2-bezylsuccinic acid, S-2-bezylsuccinic acid-4-methyl ester, methyl (2S)-2-benzyl-3-(cis-hexahydroisoindolin-2-ylcarbonyl) propionate;methodological study of content determination of impurities were all up to the requirement. The linear range of impurity A,B,C were 0.387 5-3.875,0.395-3.95 and 0.392 5-3.925 μg/ml(all r were 1.000 0). The response value of impurity A,B,C and mitiglinide calcium were 2.316 1,2.636 1,2.617 8 and 2.620 4,re-spectively. CONCLUSIONS:The structures of 3 principal degraded impurities of mitiglinide calcium have been identified and con-firmed;the content of them can be determined by HPLC main component self-comparison method.

Objective To identify the pathogens that cause hand, foot and mouth disease (HFMD) in adults and analyze the nucleotide sequences characteristics of enterovirus 71 (EV71). Methods The reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the enterovirus from the samples of four adult HFMD patients. The 227 bp amplified segments of EVT1 were then sequenced and compared with the sequences of previously isolated EVT1 strains available from GenBank by homogeneity and phylogenetic tree analyses. Results All the results of RT-PCR with enterovirus universal primers and EVT1 specific primers were positive. The EV71 sequences analysis showed that the four new sequences (named as GZ19610, GZ99310, GZ99355 and GZ46477) shared 96.0% to 99.1% nucleotide identify themselves and shared 96.9% to 100.0% homology with the strain Fuyang/17.08/3 isolated in 2008 from Fuyang, Anhui Province. Phylogenetic tree analysis showed that the genotype of the four new sequences was all subtype C4, they were the same sub-genotype as those strains isolated from Chinese mainland and Chinese Taiwan in 2004, and the genetic distance between them was most closely. Conclusions EV71 can cause adult HFMD. Compared with the nucleotide sequences of EV71 strains that isolated now and formerly in China, there is no large variation of the EV71 sequences isolated from four adult HFMD patients in Guangzhou this time. The adult HFMD patients should be isolated for treatment to avoid them transmitting the virus and causing disease spreading.

AIM:To study the action of nucleoside diphosphate kinase A(NDPK-A)on the growth of S_ 180,H_ 22,Lewis and H460.METHODS:S_ 180 or H_ 22 cell(5?106)were inoculate subcutaneously into the right armpit of 85 Kunming mice,which were randomized into 8 groups.Lewis lung carcinoma cells(2?105)were inoculate subcutaneously into the right armpit of 85 C57BL/6 mice,which were randomized as Kunming mice.From the 2nd day,the treated groups were given different dose of rhNDPK-A once a day for 8 days(for S_ 180 or H_ 22 by iv)or for 10 days(for Lewis by ip),and the control group was given physiological saline only.H460 tissue pieces about 1.5 mm?1.5 mm?1.5 mm each were inoculated subcutaneously into the armpit of 38 Balb/c/neu mice.After the volume of xenograft become 100 mm?100 mm?100 mm,the nude mice were randomized into 5 groups and given different dose of rhNDPK-A once a day for 17 days.2 days after above treatments,the mice were killed and dissected.The knubs were peeled off and weighted.RESULTS:The growth of S_ 180,H_ 22 and H460 were inhibited by rhNDPK and the growth of H_ 22 was inhibited by rhNDPK at dose of 20 mg/kg combined with cisplatin(0.5 mg/kg).But the growth of Lewis lung cancer was not inhibited.CONCLUSION:rhNDPK-A inhibited the growth of S_ 180,H_ 22 and H460.rhNDPK-A(20 mg/kg)potentiated the antitumor action of cisplatin on H_ 22.

OBJECTIVE:To optimize the parameters of the extraction and purification technologies of total flavonoids from Di-ospyros kaki Thunb. leaves,and then to compare the contents of flavonoids in fresh and dried D. kaki Thunb. leaves. METHODS:Taking the yield of total flavonoids as the investigated index,the influences of liquid-solid ratio,the volume fraction of ethanol and ultrasonic extraction time on the extraction effect were discussed by single-factor and orthogonal test. With the purity of total flavo-noids as the investigated index,the purification effects of such three kinds of methods as ethyl acetate extraction method,alcohol deposition method and macroreticular resin purification method combined with petroleum ether degreasing on the extracted solution were compared. The optimal extraction technology was adopted to extract the total flavonoids from fresh and dried D. kaki Thunb. leaves and their contents were compared. RESULTS:The optimal extraction technology was as follows as liquid-solid ratio of 25∶1 (ml/g),volume fraction of ethanol of 70%,ultrasonic extraction time of 30 min,extraction temperature of 30 ℃. The results of the verification tests showed the average content of total flavonoids was 1.75%(RSD=2.00%,n=3). The total flavonoids in the extracted solution purified by the above-mentioned three purification methods had a purity of 24.92%,15.94% and 35.52% respec-tively,in which the macroreticular resin purification method with petroleum ether degreasing combined with AB-8 resin purification was optimal. The content of flavonoids in fresh D. kaki Thunb. leaves(1.75%) was about twice as much as that in dried leaves (0.87%). CONCLUSIONS:The optimal extraction and purification technologies are simple with good effect,and suitable for large-scale production. Fresh D. kaki Thunb. leaves should be used as raw materials for extracting flavonoids.

Objective To investigate the epidemiological, clinical and pathogenic characteristics and prognosis of the first imported case of Chikungunya fever in China in 2008. Methods Epidemiological and clinical data of this mate adult patient were analyzed retrospectively. Chikungunya virus (CHIKV)-IgM was detected using enzyme linked immunosorbent assay (ELISA) and colloidal gold immunoassay. CHIKV-RNA was detected using real-time fluorescent polymerase chain reaction (RT-PCR). Results This patient had onset of fever on March 2 in 2008 and lasted for 5 days. In addition, he felt joint and muscle pains, headache and found generalized engorged maculopapule. The laboratory tests showed leukopenia and thrombocytopenia. CHIKV-IgM was detected positive at day 9 after the onset and CHIKV-RNA was all positive at day 3, 5, 7, 9 after the onset. A 575 bp fragment of RT-PCR product was sequenced and detected the nucleotide homology was 99% compared with CHIKV sequences in GenBank. The patient recovered with symptomatic supportive treatment.Conclusion This imported case of Chikungunya virus infection is reported for the first time in China.It shows similar clinical manifestations with dengue fever.

Objective:To optimize the formula composition of rutin self-microemulsifying drug delivery system by pseudo-ternary phase diagram.Methods:The oil, surfactant and co-surfactant were chosen by the solubility test , and the formula of rutin self-microe-mulsifying drug delivery system was optimized by pseudo-ternary phase diagram .Results: The formula of rutin self-microemulsifying drug delivery system was with the mass ratio of oleic acid , Cremopher RH40 and absolute ethyl alcohol of 23∶36∶12 .The microemul-sion was clear transparent liquid .Conclusion:The prepared rutin self-microemulsifying drug delivery system using the optimized for-mula screened by pseudo-ternary phase diagram can increase the solubility of rutin significantly .

Objective To evaluate the internal reliability of Psychosocial Function Evaluation for mental disabled people in day care unit. Methods 16 participants were evaluated twice by the same rater (social worker) in the unit at the Permanent stage and the Steering stage. Results Cronbach's α was greater than 0.7 in the both evaluation. The Spearman correlation coefficient between each dimension and total score were between 0.502 and 0.869. Conclusion The internal reliability of Psychosocial Function Evaluation is satisfactory for mental disabilities.

Objective To detect and analyze the haemagglutinin (HA) gene of the first influenza A-H1N1 viral strain isolated in Guangdong Province during an influenza A pandemic in 2009.Methods A-H1N1 virus strain was isolated from the throat swab of the first patient diagnosed with A-H1N1 virus infection in Guangdong Province in 2009. Viral nucleonic acid was extracted from supernatant of cell culture and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) with HA gene-specific primers. The product was cloned, sequenced, and the homology was analyzed. Results A 1710 bp HA gene of the first influenza A-H1N1 viral strain in Guangdong Province in 2009 was acquired, which was named as A/GuangzhouSB/01/2009 (H1N1) HA with GenBank access No. GQ268003. The homology of the studied HA gene and the 277 influenza A (H1N1) isolates reported in the epidemic areas was 99.0%-99.8%, and as high as 99.8% when compared with the isolates reported in the United States where the patient had traveled. When the studied HA gene was compared with 25 isolates of Chinese seasonal A-H1N1 virus, the homology was 72.3%-85.6%. Conclusions The homology of the first isolated A-H1N1 viral strain in Guangdong Province in 2009 and epidemic influenza A-H1N1 virus is high, while it is low compared with Chinese seasonal A-H1N1 virus.

Endoplasmic reticulum plays a key role in both basic structure formation and function performance of microenviron-ment. Endoplasmic reticulum homeostasis unbalance caused by endoplasmic reticulum stress has become a hot research topic in recent years. This paper focuses on the role of endoplasmic retic-ulum stress in ischemic stroke. Research progress of related sig-naling pathways were reviewed, especially mechanisms through which endoplasmic reticulum stress trigger the inflammatory reac-tion, so as to provide a new research method for prevention of is-chemic stroke.