OBJECTIVE:To prepare Levetiracetam soluble tablets and evaluate its quality. METHODS:The liquidity indexes of raw material(bulk density,tap density,etc.)and the particle size distribution,liquidity indexes,moisture of the intermediate,that was mixed granules,were investigated to establish preparation method of tablet. The content of levetiracetam was determined by HPLC,and quality indexes of soluble tablets were evaluated,such as appearance,disintegration time,the content of main com-ponent. RESULTS:The wet granulation method was established to prepare Levetiracetam soluble tablets. The specification was 100 mg/220 mg of prepared tablets,it was bright in appearance,and disintegration time was less than 1 min;average content of leveti-racetam was 100.1%,and rigidity was 7.5 kg;the friability of prepared tablets was up to the standard. CONCLUSIONS:The for-mulation and preparation technology of Levetiracetam soluble tablets are rational and controllable. The quality indexes of prepared tablets are all up to the requirements.

OBJECTIVE:To study inhibitory effects in vivo and in vitro of gemcitabine on liver cancer. METHODS:MCF-7 cells and HepG2 cells were treated with different concentrations of gemcitabine solutions(5,10,20,30,50,70 and 90 μmol/L). The absorbance of cells was determined by MTT assay after treated for 24,48 and 72 h. The inhibition ratio and 50% inhibiting concentration(IC50) of cells were calculated. Tumor-bearing mice were established by inoculating 0.2 ml liver cancer H22 cell line via right anterior axillary,and then randomly divided into control group(normal saline)and gemcitabine group(40 mg/kg)with 5 mice in each group. They were given relevant medicine intravenously every 2 days,for 3 times. The changes of body weight and in-hibition ratios were recorded. RESULTS:Gemcitabine can inhibit MCF-7 cells and HepG2 cells,and IC50 of gemcitabine to them were 30 and >90 μmol/L within 24 h respectively,and those of gemcitabine to them were all lower than 5 μmol/L after 48 h and 72 h. There was no statistical significance in body weight of tumor-bearing mice between 2 groups,and inhibitory rate of gemcitabi-ne to H22 cell line was 75.76%. CONCLUSIONS:Gemcitabine can inhibit liver cancer in vivo and in vitro.

Objective To study the effects of dibutyl phthalate(DBP) on estradiol biosynthesis and the related mechanism in puberty female rats.Methods Thirty-two female SD rats aged 21 days were randomly divided into the control group(corn oil) and low-dose(250 mg/kg),moderate-dose(500 mg/kg),and high-dose(1 000 mg/kg) DBP exposed groups,the rats were treated with DBP through gavage with a dose of 5 ml/kg,once a day for 8 consecutive weeks.After 8 weeks of exposure,the animals were sacrificed at proestrus.Radioimmunoassay was used to detect estradiol in the serum,the expression of aromatase mRNA levels was detected by real-time fluorescent quantitative RT-PCR.Results Compared with the control group,the body weight of rats in the moderate-dose group was significantly higher(P

Purpose/Significance To review the knowledge discovery methods based on knowledge graph in the biomedical field,and to provide references for researchers.Methods/Process The paper summarizes the knowledge discovery methods based on knowledge graph by systematically searching and analyzing the relevant literatures,and compares the advantages and disadvantages of various knowl-edge discovery methods.It points out that the limitations and challenges of knowledge discovery methods based on knowledge graph in the biomedical field,and puts forward suggestions and prospects.Result/Conclusion In the future research,it is suggested to improve the in-terpretability of knowledge discovery results,build an effective result evaluation framework,and establish a standardized knowledge dis-covery process with multi-domain experts'collaboration,so as to improve the quality and efficiency of knowledge discovery research.

Objective:Evaluate the application of Fourier transform infrared spectroscopy in the identification of homology of carbapenem-resistant Escherichia coli(CREC). Methods:A total of 26 carbapenem-resistant Escherichia coli strains were isolated from 9 provinces in China in 2018. The 900-1 200 cm -1 was selected as a spectral region for the Euclidean distance calculating and average linkage clustering between all isolates.The single nucleotide polymorphism (SNP) was analyzed by whole genome sequencing (WGS). Results:Twenty-six CREC strains were divided into 14 infrared spectros copy(IR) types by FTIR. The same IR type belonged to the same sequence type type.Compared with cluster analysis based on WGS, the consistency of FTIR cluster analysis was 92.3% (24/26).Conclusions:FTIR presented excellent performance in identification of homology of CREC.Besides, with the advantages of simple operation and rapid acquisition of results, FTIR may be a useful tool in clinical labs.

Objective: Schizophrenia (SCZ) is a severe psychiatric disorder with unknown etiology and lacking specific biomarkers. Herein, we aimed to explore plasma biomarkers relevant to SCZ using targeted metabolomics. Methods: Sixty drug-naïve SCZ patients and 36 healthy controls were recruited. Psychotic symptoms were assessed using the Positive and Negative Syndrome Scale. We analyzed the levels of 271 metabolites in plasma samples from all subjects using targeted metabolomics, and identified metabolites that differed significantly between the two groups. Then we evaluated the diagnostic power of the metabolites based on receiver operating characteristic curves, and explored metabolites associated with the psychotic symptoms in SCZ patients. Results: Twenty-six metabolites showed significant differences between SCZ patients and healthy controls. Among them, 12 metabolites were phosphatidylcholines and cortisol, ceramide (d18:1/22:0), acetylcarnitine, and γ-aminobutyric acid, which could significantly distinguish SCZ from healthy controls with the area under the curve (AUC) above 0.7. Further, a panel consisting of the above 4 metabolites had an excellent performance with an AUC of 0.867. In SCZ patients, phosphatidylcholines were positively related with positive symptoms, and cholic acid was positively associated with negative symptoms. Conclusion Our study provides insights into the metabolite alterations associated with SCZ and potential biomarkers for its diagnosis and symptom severity assessment.

OBJECTIVE：To study the effects of berberine on mic e macrophage polarization based on TLR 4-MyD88-NF-κB signaling pathway. METHODS ：Using mice RAW 264.7 macrophage as the object ，atorvastatin calcium as positive control ， inflammatory cell model was induced by lipopolysaccharide （LPS）；ELISA method was used to detect the contents of TNF-α，IL-6 and NF-κB in cell culture medium after treated with low，medium and high doses of berberine （5，10，20 μmol/L）for 24 h. The real-time fluorescence quantitative PCR was conducted to determine the mRNA expression of TLR 4 and MyD 88 in cells. Western blotting assay was used to detect the protein expression of TLR 4，MyD88，iNOS and CD 206 in cells. RESULTS ：Compared with blank control group ，the contents of TNF-α，IL-6 and NF-κB in cell culture medium，mRNA expression of TLR 4 and MyD 88， protein expression of TLR 4，MyD88 and iNOS in cells were increased significantly in LPS induction group （P＜0.05）. Compared with LPS induction group ，the contents of TNF-α and IL-6，mRNA and protein expression of TLR 4 and MyD 88 in atorvastatin calcium group ，berberine medium-dose and high-dose groupsas well as the content of NF-κ B and protein expression of iNOS in administration groups were decreased significantly ， while the content of NF-κB in berberine high-dose group was significantly lower than atorvastatin calcium group （P＜0.05）. The protein expressions of CD206 in atorvastatin calcium group and berberine high-dose group were increased significantly ，while the protein expression of CD 206 in berberine high-dose group was significantly higher than atorvastatin calcium group （P＜0.05）. CONCLUSIONS ：Different doses of berberine can intervene in mice macrophage polarization to different extents ，the mechanism of which may be associated with the regulation of TLR4/MyD88/NF-κB signaling pathway.

OBJECTIVETo investigate pathogens and molecular-epidemiology characteristics of viral meningoencephalitis in the monitoring sites of Zhejiang province, 2013.

METHODSCerebrospinal fluid and/or stool specimens were collected from suspected patients admitted to the monitoring hospitals in southern and northern Zhejiang province. Such specimen were subject to real-time qPCR for the detection of Human enterovirus (HEV), Japanese encephalitis virus (JEV), Mumps virus (MuV), Herpes simplex virus (HSV) and Cytomegalovirus (CMV). HEVs were isolated using the RD and Hep-2 cell lines, while VP1 genes from all HEV-positive isolates or RNA-positive specimen were amplified, sequenced, for homology and evolution analysis.

RESULTS92 (38.5%) of the 239 samples collected from 229 patients were detected as virus nucleic acid positive, including 87 HEV positive samples, 1 MuV positive, 2 HSV positive, and 2 CMV positive; of the 87 HEV positive samples, 38 were further determined to be Coxsackievirus (CV) and 49 as Echovirus (E). 56 HEV strains were isolated from 239 (23.4%) samples. From the 31 cerebral fluid specimen of nucleic acid positive yet virus isolation negative, the most specimen were identified with E9 (9 specimen), followed by CVA9 (8 specimen); the viral serotype of Zhejiang province HEV were CVA9, CVB4, CVB5, E6, E7, E9, E11, E14, E16, E25 and E30, respectively. Predominant epidemic strains identified at southern and northern Zhejiang province were CVB5 and E6 respectively. The phylogenetic analysis of VP1 gene showed that all the HEV isolates in Zhejiang province were HEV-B.

CONCLUSIONThe HEV-B was the main pathogen for viral meningoencephalitis in Zhejiang province in 2013, including 11 serotypes, while E7 was the first time to be isolated in Zhejiang province. The predominant isolates were CVB5 and E6 in southern and northern Zhejiang province respectively. The positive rate of viral nucleic acid detection was significantly higher than that of viral isolation. Regular EV isolation method was exposed to the risk of missing-detection of E9 and CVA9.

Biological Evolution ; China ; epidemiology ; Cytomegalovirus ; Encephalitis Virus, Japanese ; Encephalitis, Viral ; Enterovirus ; Enterovirus B, Human ; Hepatitis E virus ; Humans ; Meningitis, Viral ; epidemiology ; genetics ; Meningoencephalitis ; Molecular Epidemiology ; Mumps virus ; Phylogeny

OBJECTIVE： To establish HPLC fingerprints of Nauclea officinalis extract syrup， and to determine the contents of 9 components. METHODS： HPLC method was adopted. The determination was performed on Diamonsil C18（2）column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using strictosamide as reference， HPLC chromatograms of 20 batches of N. officinalis extract syrup were drawn. The similarity of HPLC chromatograms were evaluated by using TCM Fingerprint Similarity Evaluation System （2004A edition） to confirm common peaks. The contents of 9 components were determined by standard curves. RESULTS： There were 26 common peaks in 20 batches of HPLC chromatograms， and the similarity was higher than 0.98. Compared with mixed control， 9 chemical components were identified， such as 3，4-dihydroxybenzoic acid， neochlorogenic acid， loganic acid， chlorogenic acid， cryptochlorogenic acid， swertioside， pumiloside， strictosamide and vincosamide. The linear range of 9 components were 17.24-275.84， 7.56-120.96， 15.40-246.40， 7.84-125.44， 8.64-138.24， 7.96-127.36， 8.40-134.40， 48.56-776.96， 4.16-66.56 μg/mL（all r≥0. 999）， respectively. The limits of detection were 0.043 1， 0.126 0， 0.038 5， 0.130 7， 0.144 0， 0.066 3， 0.070 0， 0.012 1， 0.052 0 μg/mL， respectively. The limits of quantitation were 0.215 5， 0.189 0， 0.077 0， 0.196 0， 0.288 0， 0.132 7， 0.105 0， 0.097 6， 0.138 7 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 99.6％、106.3％、100.1％、102.0％、98.4％、100.0％、99.3％、100.6％ and 101.2％， and RSDs were 1.20％、0.24％、0.59％、1.00％、0.73％、1.30％、1.10％、1.80％、1.90％（n＝6）. CONCLUSIONS： Established HPLC fingerprints and quantitative determination method of N. officinalis extract syrup are accurate， specific and sensitive. It can provides reference for quality control of N. officinalis extract syrup.

ObjectiveTo explore the clinical efficacy of compound Wufengcao liquid （CWL） on tuberculous ulcer and the influence on macrophage polarization. Method① Clinical experiment： A total of 145 patients with tuberculous ulcer who were treated in Nanjing Integrated Traditional Chinese and Western Medicine Hospital were randomized into observation group， control group Ⅰ， and control group Ⅱ according to the random number table method. In addition to the basic anti-tuberculosis chemotherapy， CWL， Kangfuxin liquid， and isoniazid solution （local external application） were respectively used in the observation group， control group Ⅰ， and control group Ⅱ. The treatment lasted 4 weeks for each group. The total effective rate in wound healing， traditional Chinese medicine（TCM） syndrome score， and histopathological morphology of wound were observed and the expression of inducible nitric oxide synthase （iNOS） and arginase-1 （Arg-1） in wound tissue was measured. ② Cell experiment： RAW264.7 cells were cultured in DMEM （10% fetal bovine serum， 1% double-antibody solution） in a cell incubator （37 °C， 5% CO₂）. Phorbol 12-myristate 13-acetate （PMA） was used to induce the differentiation of RAW264.7 cells into macrophages. Lipopolysaccharide （LPS） was employed to stimulate polarization of macrophages into M1 type and interleukin-4 （IL-4） to induce the polarization into M2 type. Kangfuxin solution， isoniazid solution， and CWL were respectively applied to the above cell model for 36 h. The cell supernatant was collected and centrifuged. Western blot was used to detect the protein expression of tumor necrosis factor-α （TNF-α）， transforming growth factor-β （TGF-β）， iNOS， and Arg-1， and flow cytometry （FCM） to detect the expression of CD86 and CD206. Result①Clinical experiment： The total effective rate in the CWL group ［98.0% （48/49）］ was higher than that in the control group Ⅰ ［87.5% （42/48）， χ²=3.962， P<0.05］ and control group Ⅱ ［83.3% （40/48）， χ²=6.162， P<0.05］. After 28 days of treatment， compared with control group Ⅰ and control group Ⅱ， CWL decreased the TCM syndrome score （P<0.05） and obviously improved the histopathological morphology of the wound. Immunohistochemistry results showed that the iNOS expression in local focus tissue was lower （P<0.05） and the expression of Arg-1 was higher （P<0.05， P<0.01） in the CWL group than in the control group Ⅰ and control group Ⅱ after 28 days of treatment. ② Cell experiment： Western blot assay showed that the expression of iNOS and TNF-α in LPS group increased compared with that in the M0 group （P<0.01） and the expression in the LPS+ isoniazid group， LPS+ Kangfuxin group， and LPS+CWL group was lower than that in the LPS group （P<0.05）. The expression of iNOS in LPS+Kangfuxin group and LPS+ CWL group was lower than that in the LPS+isoniazid group （P<0.05， P<0.01）， and the expression of TNF-α in LPS+ CWL group was lower than that in LPS+isoniazid group （P<0.01）. The expression of TNF-α in LPS+ CWL group decreased compared with that in the LPS+ Kangfuxin group （P<0.05）. The expression of Arg-1 and TGF-β in IL-4 group was higher than that in the M0 group （P<0.01）， and the expression in the IL-4+isoniazid group， IL-4+Kangfuxin group， and IL-4+ CWL group was higher than that in the IL-4 group （P<0.05）. The expression of Arg-1 and TGF-β in the IL-4+ Kangfuxin group and IL-4+CWL group was higher than that in the IL-4+isoniazid group （P<0.05， P<0.01）， and the expression was higher in the IL-4+CWL group than in the IL-4+Kangfuxin group （P<0.05， P<0.01）. The FCM result showed that the expression of CD86 and CD206 in LPS group and IL-4 group was higher than that in M0 group （P<0.01）. CD86 expression in LPS+isoniazid group， LPS+ Kangfuxin group， and LPS+CWL group was lower than that in the LPS group （P<0.01）. The expression of CD86 in LPS+Kangfuxin group and LPS+ CWL group increased compared with that in the LPS+isoniazid group （P<0.01）， and the expression was higher in the LPS+ CWL group than in the LPS+Kangfuxin group （P<0.01）. CD206 expression in IL-4+ isoniazid group， IL-4+Kangfuxin liquor group， and IL-4+ CWL group was increased compared with that in the IL-4 group （P<0.01）. CD206 expression in IL-4+Kangfuxin liquid group and IL-4+ CWL group was decreased compared with that in the IL-4+isoniazid group （P<0.01）. CD206 expression in IL-4+CWL group was lower than that in the IL-4+ Kangfuxin group （P<0.05）. ConclusionCWL can promote the healing of tuberculous ulcers， and the mechanism is that it inhibits the expression of iNOS， TNF-α， and CD86 and promotes the expression of Arg-1， TGF-β， and CD206， thereby regulating M1/M2 polarization balance.