Objective To avoid false positive detection ofVibrio cholerae and improve the detection correct rate.Methods 1~7 months of 2013 were randomly selected,the national various provinces and cities CDC to China cholera CDC positive screening 14 strains.LB nutrient agar 12 hours,take single colony to Vibrio cholera serum agglutination,extraction of strain DNA at the same time boiled template method.For Vibrio 16SrDNA sequence and design primers for PCR detection of Vib-rio,16SrDNA,electrophoresis were used to observe the 16SrDNA products,16SrDNA positive products sent to sequencing company sequencing,sequencing results were Blast comparison on the NCBI website for the analysis and comparison of ser-um agglutination and Blast alignment.Results 12 strains was positive for agglutination and 2 strains of non agglutination in 14 strains.The Vibrio 16SrDNA amplification,electrophoresis were used to observe the 14 isolates that were amplified frag-ment corresponding,that the selected strains were vibrio.The 16SrDNA positive products were 14 strains,and the sequen-cing the Blast results:2 strains of bacteria were Vibrio harveyi for non agglutination,in 12 positive strains of serum aggluti-nation;1 strains was Vibrio natriegen and 11 strains wereVibrio cholerae .Conclusion Detection ofVibrio cholerae cholerae combined with serum agglutination and gene sequencing can avoid false positive result ofVibrio cholerae ,and improve cor-rect rate of the detection.

[ABSTRACT]OBJECTIVETo investigate the therapeutic effect of early combination treatment with intratympanic glucocorticoid injection for sudden sensorineural hearing loss(SSNHL).METHODSTotal of 178 patients with diagnosis of SSNHL, including 44 with hearing loss at middle-low frequencies,50 at high frequencies,44 at all frequencies and 40 at total deafness, were randomly divided into combination treatment group (CT group) and systemic oral prednisone group (OP group).CT group (82 patients) received intratympanic administration of methylprednisolone and oral prednisone in tapering doses. Pure-tone threshold audiometry (PTA) was performed at week 8 after beginning of therapy. RESULTSIn middle-low frequencies, the final PTA and PTA improvement were 26.14±24.82 dB and 42.23±1.61 dB in CT group respectively, and were 37.91±13.98 dB and 30.00±13.30 dB in OP group respectively (both P<0.05). In high frequencies, all frequencies and total deafness SSNHL group, the differences of the final PTA and PTA improvement were not statistically significant in both groups. The PTA improvement in CT group for total deafness (26.52±14.03) dB was more than that of OP group (22.06±11.17)dB (t=2.361,P<0.05). In middle-low frequencies, the recovery rate was 72.73%(16 cases) in CT group, and 40.91% (9 cases) in OP group, the difference was significant (χ2=4.539,P<0.05), but not in apparent effective rate, effective rate and total effective rate. In high frequencies, all frequencies and total deafness, there were no significant differences in therapeutic effect. The recovery rate of total CT group (40.24%) was significantly increased compared with that of OP group (26.04%)(χ2=4.061,P<0.05), the differences of apparent effective rate, effective rate and total effective rate were not statistically significant in both groups.CONCLUSIONFor low-middle frequencies SSNHL, the early combination treatment with intratympanic glucocorticoid injection leads to higher hearing recovery and therapeutic effect compared with traditional oral prednisone. Systemic application should be applied for high frequencies, all frequencies and total deafness of SSNHL.

Objective To investigate neutrophil lymphocyte ratio (NLR) in early-stage diabetic nephropathy and its clinic significance. Methods The 145 subjects were divided into two groups:the healthy control group (n=54) and early stage diabetic nephropathy group (n=91). The numbers of neutrophils (N) and lymphocytes (L) as well as the NLR values of peripheral blood and other biochemistry index were examined. Factors of early stage diabetic nephropathy were calculated us?ing variance and logistic regression analysis. Results Creatinine(Cr), total cholesterol(TC), triglyceride(TG), LDL-C, neu?trophils number and CRP in DN group were significantly higher than those of the control group and lymphocytes numbers of DN group were significantly lower than that of the control group (P<0.05 respectively);NLR values were significantly higher in diabetic nephropathy group compared with those of healthy control group(2.52±0.57 vs 1.82±0.60,t=6.997, P<0.01). Lo?gistic regression analysis showed that the risk factors of DN include NLR, TG and total cholesterol. NLR ( P <0.001, OR=8.951, OR 95%CI:3.595-22.287) was significantly associated with DN. Conclusion High NLR values may be a predic?tive and reliable marker ofearly-stage DN.

Objective: To understand the status and relationship between bullying behaviors and subjective well-being among left-behind students in Bijie,and to provide reference for the governments to support the development of policies and intervention. Methods: A total of 735 left-behind students were investigated on the spot with the Olweus Bully Questionnaire and Adolescent Subjective Well-being Scale. Results: Among the 735 left-behind students, 28.71% were found of participating in behaviors of bullying. More boys involved behaviors of bullying (bullies,victims and bully-victims) others than the girls(χ2=25.93, P<0.05), Bullying was more likely to be reported among children with poor peer relations(χ2=12.46, P<0.05). Total scores of the feelings of friendship，family，freedom,negative emotion and life differed significantly among different bullying roles(F=12.76, 2.86, 3.30, 5.98, 4.72, P<0.05). Multivariate analysis showed that there were many factors affecting the subjective well-being of these students, such as bullying behaviors, relationship with their classmates, academic record, the frequency which they contacted with parents(P<0.05). Conclusion Bullying is highly prevalent among left-behind children in Bijie. Effective prevention and control mechanisms should be established at the social, school and family levels to reduce the occurrence of bullying. Attention should be paid to the psychological and behavioral dynamics of left-behind middle school students, so as to improve their happiness.

This article was aimed to study the chemical constituents of the petroleum ether fraction of Liparis nervosa.Chemicalcompoundswereisolatedandpurifiedthroughvariouschromatographytechniques.The accurate structures of chemical compounds were confirmed with spectral data and literatures. The results showed that7chemicalconstituentswereisolatedfromthepetroleumetherfractionofLiparis nervosa,whichwere moscatin (1), batatasin Ⅲ (2), bergapten (3), isoimpinellin (4), xanthotoxin (5), imperatorin (6) and β-sitosterol (7). It was concluded that chemical constituents 3-7 were isolated from this genus for the first time. And chemical constituents 1-7 were isolated from this plant for the first time.

Objective: To investigate risk factors and epidemiological characteristics of bacillary dysentery outbreaks in three schools, and to provide scientific basis for the prevention and control of the epidemic in the future. Methods: Case definition was established. All suspected, possible and confirmed cases of all students and faculty members from 3 schools (A, B, C) were selected for epidemiological investigation. Control group was used for case-control analysis, and relevant samples were collected for laboratory testing. Results: A total of 132 cases were found in 3 schools, all of which were from students, with the incidence rate of 17.74%. The morbidity in kindergarten A was 20.00%, in center primary school B it was 21.74%, and in junior middle school C it was 11.61%. Cohort studies and casecontrol studies suggested that schools are exposed places and that washing hands with raw water in schools was possible risk factor [OR(95%CI) =4.50(1.01-20.11)]. Nine stool samples were tested in laboratory, among which 8 were positive for Shigella(88.99%), and Shigella was detected in the end nodes of school s pipeline network, the water samples from canteen bucket, and the floor drains of sewer pipe. Conclusion The bacillary dysentery outbreaks in 3 schools was caused by Shigella, which may be due to fecal contamination of domestic water in 3 schools before the start of the school year. It is suggested to strengthen the management of centralized water supply and construction in rural areas, intensify the supervision at all levels, and sanitation and disinfection before school opens at all levels.

Purpose: Inflammatory myofibroblastic tumor (IMT) is a rare mesenchymal malignancy that occurs primarily in children and adolescents. The clinical and pathological features of IMT in adult patients are not well understood. Materials and Methods: We retrospectively searched for records of adult patients with IMT at Fudan University Shanghai Cancer Center from 2006 to 2021. Clinicopathological data, treatments, and outcomes were collected and analyzed. Results: Thirty adult patients with IMT, mostly women (60.0%), were included. The median age of the patients was 38 (21-77). The most common primary site was abdominopelvic region (53.3%), followed by lungs (20.0%). Seven patients had an abdominal epithelioid inflammatory myofibroblast sarcoma (EIMS). The positivity rate of anaplastic lymphoma kinase (ALK) was 81.5% (22/27). Sixteen patients with advanced ALK-positive disease received crizotinib, with an objective response rate (ORR) of 81.3% and a disease control rate of 87.5%. The median progression-free survival was 20.8 months. EIMS was associated with more aggressive behavior; however, the prognosis was similar to that of non-EIMS patients after treatment with an ALK inhibitor. At a median follow-up time of 30 months (95% confidence interval [CI], 13.6 to 46.4), the 5-year overall survival was 77% (95% CI, 66 to 88) in all patients. Conclusion Adult IMTs appeared more aggressive, with a higher incidence of recurrence and metastases, and patients with EIMS had more aggressive cases. Treatment with ALK inhibitors resulted in a high ORR and a durable response, which suggested that ALK inhibitors could be used as a first-line treatment option in adult patients with ALK-positive advanced IMT.

We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) gene family from Huperzia serrate. The two transcripts coding HDR, named HsHDR1 and HsHDR2, were discovered in the transcriptome dataset of H. serrate and were cloned by reverse transcription-polymerase chain reaction (RT-PCR). The physicochemical properties, protein domains, protein secondary structure, and 3D structure of the putative HsHDR1 and HsHDR2 proteins were analyzed. The full-length cDNA of the HsHDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids, whereas the HsHDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids. These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (PF02401), but without the transmembrane region and signal peptide. The most abundant expression of HsHDR1 and HsHDR2 was detected in H. serrate roots, followed by the stems and leaves. Our results provide a foundation for exploring the function of HsHDR1 and HsHDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.

【Objective】 To investigate the quality changes of suspended red blood cells (SRBCs) prepared from the blood of Tibetan high Hb population, and explore the availability and safety of blood collected from Tibetan high Hb population. 【Methods】 The voluntary blood donors were grouped according to the Hb concentration at the initial screening: female blood donors from Tibet Autonomous Region (>3 500 m) with Hb≥190 g/L and male blood donors with Hb≥210 g/L were classified as plateau high hemoglobin group. A total of 13 male blood donors from Tibet Autonomous Region were recruited. And the female blood donors (n=13) with Hb(115~165) g/L and male blood donors (n=12) with Hb(120~185) g/L from Chengdu were classified as control group. Whole blood of 200 mL specification was centrifuged to remove the plasma, and MAP additive solution was added to prepare SRBCs, then SRBCs were divided into four aliquots (50 mL/bag and stored at 4℃. Parameters as blood routine, free Hb and hemolysis rate were measured aseptically at day 1, 14, 21, 35 of storage. And 10 mL SRBCs was used to extract membrane proteins for tyrosine phosphorylation detection of band 3 protein. 【Results】 The RBCs counts(×10¹²/L), hematocrit(%) and hemoglobin(g/L) of Tibetan high Hb group and control group were 6.76±0.95 vs 4.65±0.52, 63.3±6.8 vs 43.1±4.4 and 214.4±19.8 vs 143.2±16.9 (P<0.01). The erythrocyte deformability test on the day 1, 14, 21, 35 of storage showed that the deformability of SRBCs prepared from Tibetan high Hb group was significantly lower than that of the control group under shear stress of 3, 5.33, 9.49, 16.87, and 30 Pa, while the hemolysis rate of SRBCs prepared from the Tibetan high Hb group and the control group on the day 1, 14, 21, 35 were 0.050 2±0.040 2 vs 0.022 2±0.011 1, 0.055 4±0.043 vs 0.032 1±0.028 7, 0.061 2±0.025 9 vs 0.034 3±0.031 7 and 0.069 6±0.032 0 vs 0.044 0±0.033 3 (P<0.05). Western blotting showed that the cytoplasmic N-terminal Y21 of band 3 protein of SRBCs prepared from Tibetan high Hb group was highly phosphorylated. 【Conclusion】 The deformability of SRBCs prepared from the Tibetan high Hb group was significantly lower while the hemolysis rate of SRBCs was higher than that of the control group. The hemolysis rate of the SRBCs at the end of storage prepared from the Tibetan high Hb group meets the requirements of the national standard GB18469-2012(<0.8%). The increase of hemolysis rate of SRBCs prepared from the Tibetan high Hb group was closely related to the phosphorylation of band 3 protein.

Medicinal plants are a valuable source of essential medicines and herbal products for healthcare and disease therapy. Compared with chemical synthesis and extraction, the biosynthesis of natural products is a very promising alternative for the successful conservation of medicinal plants, and its rapid development will greatly facilitate the conservation and sustainable utilization of medicinal plants. Here, we summarize the advances in strategies and methods concerning the biosynthesis and production of natural products of medicinal plants. The strategies and methods mainly include genetic engineering, plant cell culture engineering, metabolic engineering, and synthetic biology based on multiple "OMICS" technologies, with paradigms for the biosynthesis of terpenoids and alkaloids. We also highlight the biosynthetic approaches and discuss progress in the production of some valuable natural products, exemplifying compounds such as vindoline (alkaloid), artemisinin and paclitaxel (terpenoids), to illustrate the power of biotechnology in medicinal plants.