OBJECTIVE: To establish the characteristic chromatogram of Baphicacanthis Cusiae Rhizoma et Radix and its decoction pieces by HPLC for the identification of authentic and counterfeit products. METHODS: High performance liquid chromatography (HPLC) was used with Agilent Zorbax C18(4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-water with gradient elution. The detector was a secondary tube array (DAD). The column temperature was maintained at 35 ℃, the flow rate was 1.0 mL•min-1, and the injection volume was 10 μL. RESULTS: Fifteen batches of genuine crude drug and twelve batches of genuine decoction pieces were determined. Five common peaks were found, among which three peaks were 2-benzoxazolinone, indigo and indirubin. CONCLUSION: The established characteristic chromatogram of Baphicacanthis Cusiae Rhizoma et Radix can effectively distinguish the authentic from the counterfeit. The methodological demonstration shows that the method is accurate, stable and reproducible.

Listeriamonocytogenes is a facultative intracellular bacterium that enters professional antigen presenting cells , presents passenger antigens to the major histocompatibility complex class I and II pathways ,then elicits CD+4 and CD+8 T-cell-mediated immune responses .It was demonstrated that attenuated Listeriamonocytogenes as a novel live vaccine vector in deliv-ering tumor antigens of cervical cancer and melanoma etc .,could induce strong protective immune response ,and shows effec-tive antitumor immunotherapeutics .This review discussed the characteristics of immune responses elicited by Listeria monocy-togenes ,and the progress of its antitumor immunotherapeutics as delivery vaccine vector .

Antigen Rv2628 of Mycobacterium tuberculosis is associated with latent tuberculosis infection. In this study, Rv2628 was prokaryotic expressed and purified, its immunological characteristics was evaluated with macrophage cell line RAW264.7 and BALB/c mice. The results show that Rv2628 was mainly expressed in form of inclusion body confirmed by SDS-PAGE, and could react with rabbit anti-H37Rv polyclonal antibody detected by Western blotting assay, indicating that the protein had an effective immunoreactivity. The interactions between Rv2628 and macrophage cell line RAW264.7 confirmed that it could effectively induce cells to produce pro-inflammatory cytokines, the relative expression level of IL-6 mRNA was higher than the control group in 1-12 h. BALB/c mice were subcutaneously immunized with Rv2628 protein, the production of IFN-gamma and IL-4 in the spleen cells was determined by Sandwich ELISA, in the Rv2628 immunized group, the level of IFN-gamma was significantly higher than that of IL-4 (P < 0.000 1). It indicated the protein induced Th1-tendency immune responses. At the same time, Rv2628(11-30) peptide used as coating antigen, the murine serum antibody titer detected by indirect-ELISA was 1:1 600, which demonstrated that Rv2628 could also induce humoral immune responses. In summary, Rv2628 could induce specific pro-inflammatory cytokines, affectively induce strongly Th1-tendency immune response and humoral response, it could be a potential target for developing subunit vaccine against TB. In addition, it laid foundation for probing the cross-talk between M. tb and host.
Animals ; Antibodies, Bacterial ; blood ; Antigens, Bacterial ; immunology ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Interleukin-6 ; immunology ; Macrophages ; immunology ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; Th1 Cells ; immunology ; Tuberculosis ; immunology

Objective Training a few medical students as standardized patients to teach and test other medical students'physical examination skills is a good way to compensate the deficient teaching resources and improve the medical students'clinical capabilities.Methods We spent 12 class hours to train 6 students as standardized patients.Then we used these 6 students'standardized patients to help the teacher to educate medical students in experimental group,while students in the control group only had traditional class under the construction of the teachers as before.At last,we collected their final physical examination skills'score.Results The final physical examination skill score of experimental group(85.78?5.89)is higher than that of control group(84.53?4.64)(P

This paper is discussed about course system construction of Microbiology, teaching method, in- struction means and experimental teaching mode. Teaching practice indicated that reform the pattern of Mi- crobiology educational mode can stimulate students’ interest in studying the course, cultivate their inde- pendent ability to solve questions, develop their creative thinking. It is an important way to train high-caliber talents.

The prevalence of human and animal listeriosis for nearly 11 years in China was investigated in this study . The literature information about listeriosis in China from 2002 to 2012 was collected through retrieval system to make clinical and epidemiological statistical analysis of listeriosis .Cases of listeriosis were reported in 27 (79% ) provinces of China .The re-sult showed that animal listeriosis was reported for 123 times ,among these reports ,most were from pigs (39% ) ,and the sheep was in second place .Central nervous system infection was the main clinical manifestation of listeriosis in animals (72% ) . For human listeriosis ,84 clinical cases of listeriosis were reported ,including 35% cases in non-perinatal stage and 65% cases in perinatal stage .The main clinical manifestation of listeriosis was septicemia (51% ) .According to the result of investigation about listeriosis based on literatures information ,Listeriamonocytogenes caused humans and animals listeriosis annually ,which were reported in most provinces of China .The epidemic characteristics for listeriosis suggested that it was essential to strength-en the prevention and control of listeriosis .

OBJECTIVETo examine the incidence and clinical significance of NPM1 mutations in childhood acute myeloid leukemia (AML) patients.

METHODSNPM1 mutations of 70 newly diagnosed childhood AML were detected by high resolution melting (HRM) analysis on the LightCycler 480. The incidence and clinical significance were analyzed.

RESULTSNPM1 mutations were identified in 32 (45.7%) of the 70 AML children. There was no significant difference in clinical characteristics between patients with or without NPM1 mutation, but patients with NPM1 mutation had a higher platelet count (P = 0.013). There was also no significant difference in NPM1 mutation between normal and abnormal karyotype groups. In AML-ETO or PML-RARα positive groups, the incidence of NPM1 mutations was significant lower (P = 0.048). There was no significant difference in response rates after induction therapy (P = 0.217), but the complete remission (CR) rate was higher in the NPM1-mutated group (81.3%). There was a trend toward higher event-free survival (EFS) and overall survival (OS) rates in the NPM1 mutated patients than that in wild NPM1 patients (EFS = 53.8% vs 41.4%, OS = 52.7% vs 39.2%), but the difference was not statistically significant (P = 0.374 and 0.380).

CONCLUSIONNPM1 mutations were relatively common in our cohort of AML patients. There was no significant difference in clinical characteristics between patients with and without NPM1 mutation. The NPM1 mutation patients group seemed to have better therapy response, but the difference was not statistically significant.

HPV16 E7 fusion protein was expressed in E. coli BL21, and its applied value for HPV was evaluated. HPV16 E7 gene was amplified by PCR, and cloned into prokaryotic expression vector pGEX6p-1. The recombinant plasmid was transformed into E. coli BL21, and HPV16 E7 fusion was expressed through IPTG induction. The expressed product was analyzed by SDS-PAGE and Western blot, subsequently purified according to Glutathione Sepharose 4B purification procedure. An indirect ELISA with the purified fusion protein as the coating antigen was then established to detect E7 serum antibodies from mice immunized with recombinant Listeria monocytogenes delivering HPV16 E7. The results demonstrated that the soluble fusion protein was highly expressed at 25 degrees C after induction with 0.5 mM IPTG. Furthermore, the result of Western blot analysis showed that the fusion protein had good specific reaction with an anti-E7 monoclonal antibody. Indirect ELISA result confirmed that the fusion protein could detect the serum antibodies against E7 with a titer of 1:200. The expressed GST-E7 fusion protein was immunocompetent, which was useful in the research of E7 biological function and therapeutic vaccine.
Animals ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Female ; Mice ; Mice, Inbred C57BL ; Papillomavirus E7 Proteins ; biosynthesis ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; isolation & purification

Objective:To report embryonic testicular regression syndrome(ETRS) caused by DHX37 heterozygous variant for the first time in China and summarize the clinical manifestations of ETRS as to improve the understanding of doctors for this disease.Methods:The clinical data and whole exome sequencing results of five cases of ETRS from Shenzhen Children′s Hospital were collected. The reported cases of DHX37 heterozygous variant were reviewed.Results:Five patients with ETRS visited the doctors at the age of 2 months to 5 years and 5 months. Three patients raised as males came to hospital due to virilition and 2 female patients visited a doctor due to clitoral hypertrophy. No uterus was detected by ultrasound in all patients. The gonadal pathologies from 4 cases displayed no testicular tissue or gonadal dysgenesis, complicated with gonadoblastoma in one case. The genetic testing revealed that the heterozygous variant(c.923G>A, p. R308Q) in DHX37 was found in 2 cases, without variant in other 3 cases. According to the review, ETRS and 46, XY gonadal dysgenesis due to DHX37 herozygous variant was firstly reported in 2019. A total of 40 cases, including 21 cases of ETRS, presented with the virilition or female phenotype, with the disappearance of testicular tissue as the main pathologies. There is no report in China.Conclusion:The article summarized the clinical manifestations and whole exome sequencing results of 5 patients with ETRS, among which two cases were caused by DHX37 variants and one was complicated with gonadoblastoma.

Objective To construct a lentiviral vector for overexpression of bone morphogenetic protein 7(BMP7)in mice,and the effect of BMP7 overexpression on the expression of Jagged1 in mouse aortic endothelial cells and the calcification of the co-cultured vascular smooth muscle cells(VSMCs)were analyzed.Methods According to the target gene information Mouse-BMP7(NM_007557.3)and plasmid information pLVX-zsGreen-C1,gene sequence synthesis was carried out to construct BMP7 overexpression lentivirus.The efficiency of BMP7 overexpression lentivirus infection was detected by qPCR;the expression of Jagged1 protein in aortic endothelial cells from infected mice was detected by Western blot.The endothelial cells with lentivirus overexpressing BMP7 were co-cultured with VSMCs,and the calcification of VSMCs was observed by alizarin red staining.Results BMP7 overexpression lentiviral vector was successfully constructed and transfected into aortic endothelial cells.qPCR test results showed that the expression level of BMP7 mRNA was significantly increased in the BMP7 overexpression group than that in the normal control group(P<0.01),while there was no significant difference in the expression of BMP7 mRNA between the empty vector control group and the normal control group(P>0.05).Western blot results showed that the expression level of Jagged1 protein in endothelial cells of mouse in the BMP7 overexpression group was significantly lower than that in the normal control group(P<0.01),while there was no significant difference in the expression level of Jagged1 protein in endothelial cells between the empty vector control group and the normal control group(P>0.05).The results of alizarin red staining showed that the calcification of VSMCs was significantly increased after co-cultured with endothelial cells infected with BMP7 lentivirus.Conclusion Mouse BMP7 overexpression lentiviral vector was successfully constructed,and overexpression of BMP7 can reduce the expression of Jagged1 in mouse aortic endothelial cells and promote the calcification of co-cultured VSMCs.