Research on the clustered regularly interspaced short palindromic repeats genotyping of Yersinia pestis in the natural plague foci of Hainan Tibetan Autonomous Prefecture, Qinghai / 中国热带医学

@#Abstract: Objective　To investigate the clustered regularly interspaced short palindromic repeats (CRISPR) genotypes and regional distribution of Yersinia pestis strains in the natural plague foci of Hainan Tibetan Autonomous Prefecture of Qinghai Province (referred to as "Hainan prefecture") and provide a scientific basis for plague prevention and control in this area. Methods　A total of 36 representative Yersinia pestis strains, which were isolated from different host animals and insect vectors from 1954 to 2009 in Hainan Prefecture, were selected as experimental subjects. The DNAs were extracted using the traditional sodium dodecyl sulfate decomposition and phenol-chloroform method. Three pairs of CRISPR primers (YPa, Ypb, YPc) were used for PCR amplification, sequencing and analysis of the DNA of the tested strains, respectively, as a means to identify the CRISPR genotypes of Yersinia pestis in Hainan Prefecture. Results　A total of 17 spacers were observed among 36 strains of Yersinia pestis, including 9 of YPa, 5 of YPb and 3 of YPc. All strains were divided into 5 CRISPR gene clusters (Cb2, Cb4 ', Ca7, Ca7 ', Ca35 ') and 6 genotypes (G1, G9, G22, G22-A1 ', G26-A1 ', G26-A1 'A4 -). The G26-a1 ' was the main genotype, which was distributed in Gonghe, Guide and Xinghai County, and the G22 is the second type, which was distributed in Gonghe and Guide County. Conclusions　The genetic polymorphism of CRISPR loci of Yersinia pestis strains in Hainan was high, and the regional distribution characteristics of Yersinia pestis strains with different genotypes were significant.

Screening of rpsL mutations in streptomycin resistance gene of 104 strains of Yersinia pestis strains in south area of Qinghai Province by TaqMan-MGB fluorescent probe / 中国热带医学

@#Abstract: Objective To investigate the current status of streptomycin resistance of Yersinia pestis caused by point mutations of rpsL gene in Qinghai, so as to provide theoretical basis for precise clinical medication and prevention of drug resistance of human plague outbreak in South area of Qinghai Province in the future. Methods A total of 104 representative strains of Yersinia pestis collected from plague patients, vector insects and intermediate hosts in South area of Qinghai Province from 1957 to 2009 were screened, isolated and cultured by Hiss agar plates. The DNA of representative Yersinia pestis was extracted by sodium dodecyl sulfate lysis and phenol-chloroform method. The primers forward primer and reverse primer and TaqMan-MGB probes probe1 [FAM] and probe2 [VIC] were designed for the rpsL gene of streptomycin resistance gene in China. Real-time PCR with TaqMan-MGB fluorescent probe was used to detect the mutations of rpsL gene in streptomycin resistance locus of 104 strains of Yersinia pestis in South area of Qinghai Province. Results The FAM test results of 104 strains in South area of Qinghai Province were positive, corresponding to the detection of rpsL (128 : A ), RFU peak >1 000,negative <200. VIC test results of all tested strains were negative, corresponding to the detection of rpsL (128:G), RFU peak <200, positive >1 000. That is, no strains with rpsL gene mutation related to streptomycin resistance were found in the 104 strains of Yersinia pestis in Qingnan Province. Conclusion This study provides basic data on the distribution of streptomycin resistance of Yersinia pestis in South area of Qinghai Province, and lays a foundation for preventing the occurrence of drug resistance and clinical treatment of Yersinia pestis in South area of Qinghai Province.

Genetic study of Yersinia pestis strains isolated from the Himalayan marmot natural focus area and domestic rat plague focus area in southern China / 中国热带医学

@#Abstract: Objective To understand the phenotypic and genetic characteristics of Yersinia pestis strains isolated from Himalayan marmot natural focus area and domestic rat plague focus area in southern China, and provide reference for mastering the pathogenic characteristics of Yersinia pestis of two plague foci. Methods A total of 412 of Yersinia pestis strains isolated from Himalayan marmot plague focus and domestic rat plague focus of southern China were subjected to to sorbitol fermentation assays, virulence factor, different region (DFR) typing, and clustered regularly interspaced palindromic repeats (CRISPR) typing. Results The biochemical types of Y. pestis from the two plague foci showed distinct regional distribution features. Five biochemical phenotypes were identified in Yersinia pestis isolated from Himalayan marmot natural focus area, while only one biochemical phenotype was identified in strains isolated from the domestic rat plague focus of Southern China. Most of the Yersinia pestis isolated from the two plague foci were capable of producing the virulence factors of Fl and PstI. Among the strains from Himalayan marmot focus, 70.53% (201/285) were VW-positive, 75.09% (214/285) were Pgm-positive, 20.00% (57/285) of the strains were Pgm-negative, and 5.26% (15/285) were Pgm mixed-type strains. Among strains from domestic rat plague focus of southern China, 37.80% (48/127) were VW-positive, 29.13% (37/127) were Pgm-positive, 58.27% (74/127) were Pgm-negative, and 12.60% (16/127) were Pgm mixed-type strains. DFR typing revealed 22 genotypes of Y. pestis from the Himalayan marmot plague focus, with the main genotypes being type 5, 7, 8, 10, 19, 32 and 49. All strains from domestic rat plague focus area in southern China belonged to type 9. CRISPR typing revealed that all strains from the Himalayan marmot natural focus were classified into 7 CRISPR gene clusters and 14 CRISPR genotypes, with the main genotypes being G7, G22, G26-a1'and G22-A1'. All strains from domestic rat plague focus area in southern China belonged to CRISPR genotype G30, with the gene cluster being Ca8. Conclusions The phenotypes and genotypes of the Yersinia pestis of Himalayan marmot plague focus are diverse, with an obvious characteristics of geographical distribution. The phenotype and genotype of the Yersinia pestis of domestic rat plague focus of Southern China are single. DFR and CRISPR genotyping methods with phenotypic characteristics can effectively identify the Yersinia pestis isolated from the two plague foci, thereby meeting the needs of identification and traceability research.

RGD peptide inhibits tumor growth by affecting angiogenesis related signaling pathway of laryngeal cancer stem cells / El péptido RGD inhibe el crecimiento tumoral al afectar la vía de señalización relacionada con la angiogénesis de las células madre del cáncer de laringe

SUMMARY: This study is to investigate the role and mechanism of RGD peptide in laryngeal cancer stem cells (CSCs). Laryngeal cancer CD133+Hep-2 CSCs were sorted by flow cytometry. RGD peptide was co-cultured with sorted laryngeal CSCs. Cell proliferation was detected with CCK-8 assay. The mRNA levels of VEGF/VEGFR2/STAT 3/HIF-1α were detected with RT-PCR. The proteins of VEGF/ VEGFR2/STAT 3/HIF-1α were detected with Western blot. The sorted CSCs were inoculated into nude mice. Tumor volume was measured. Integrin αvβ3 expression in tumor tissues was analyzed with immunohistochemistry. The results showed that the ratio of CD133+ CSCs to the total number of cells was 1.34±0.87 %, while CD133-non-tumor stem cells accounted for 95.0±5.76 %. The sorted cancer stem cells grew well. The RGD peptide significantly inhibited the proliferation of CD133+Hep-2 laryngeal CSCs in a dose-dependent manner. The RGD peptide significantly inhibited the mRNA of VEGFR2, STAT3 and HIF-1α in laryngeal CSCs in a concentration-dependent manner. Consistently, the RGD peptide significantly inhibited the protein expression of VEGFR2, STAT3 and HIF-1α in laryngeal CSCs in a dose-dependent manner. At the same time, in vivo tumor experiments showed that the RGD peptide significantly inhibited tumor volume but not the body weight. Furthermore, RGD peptide significantly inhibited the expression of tumor angiogenesis-related protein integrin αvβ3. Our findings demonstrate that RGD peptide inhibits tumor cell proliferation and tumor growth. The underlying mechanism may that RGD inhibits tumor angiogenesis-related signaling pathways, thus affecting the tumor angiogenesis, and decreasing the progression of human laryngeal CSCs.

Este estudio se realizó para investigar el papel y el mecanismo del péptido RGD en las células madre del cáncer de laringe (CSC). Las CSC CD133+Hep-2 de cáncer de laringe se clasificaron mediante citometría de flujo. El péptido RGD se cocultivó con CSC laríngeas clasificadas. La proliferación celular se detectó con el ensayo CCK-8. Los niveles de ARNm de VEGF/VEGFR2/ STAT 3/HIF-1α se detectaron con RT-PCR. Las proteínas de VEGF/ VEGFR2/STAT 3/HIF-1α se detectaron con Western blot. Las CSC clasificadas se inocularon en ratones nudos. Se midió el volumen del tumor. La expresión de integrina αvβ3 en tejidos tumorales se analizó con inmunohistoquímica. Los resultados mostraron que la proporción de CSC CD133+ con respecto al número total de células fue de 1,34 ± 0,87 %, mientras que las células madre no tumorales CD133 representaron el 95,0 ± 5,76 %. Las células madre cancerosas clasificadas crecieron bien. El péptido RGD inhibió significativamente la proliferación de CSC laríngeas CD133+Hep-2 de una manera dependiente de la dosis. El péptido RGD inhibió significativamente el ARNm de VEGFR2, STAT3 y HIF-1α en CSC laríngeas de manera dependiente de la concentración. De manera consistente, el péptido RGD inhibió significativamente la expresión proteica de VEGFR2, STAT3 y HIF-1α en CSC laríngeas, de manera dependiente de la dosis. Al mismo tiempo, los experimentos con tumores in vivo mostraron que el péptido RGD inhibía significativamente el volumen del tumor pero no el peso corporal. Además, el péptido RGD inhibió significativamente la expresión de la proteína integrina αvβ3 relacionada con la angiogénesis tumoral. Nuestros hallazgos demuestran que el péptido RGD inhibe la proliferación de células tumorales y el crecimiento tumoral. El mecanismo subyacente puede ser que RGD inhiba las vías de señalización relacionadas con la angiogénesis tumoral, afectando así la angiogénesis tumoral y disminuyendo la progresión de las CSC laríngeas humanas.

Research Progress on Metabolite Identification of Synthetic Cannabinoid New Psychoactive Substances / 法医学杂志

Synthetic cannabinoids are currently a class of new psychoactive substances with the largest variety and most abused. Metabolite identification research can provide basic data for monitoring synthetic cannabinoids abuse, which is the current research hotspot. The main trend of structural modification of synthetic cannabinoid is to replace the fluorine atom on pentyl indole or indazole cyclopentyl with hydrogen atom, which greatly improves the biological activity of the compound. The main metabolic reactions include hydroxylation, fluoropentyl oxidative, ester hydrolyze, amide hydrolysis. Liquid chromatography-high resolution mass spectrometry has become the preferred choice for the structural identification of metabolites. This review mainly summarizes research on metabolism software prediction and human hepatocyte model, human liver microsomes model, rat in vivo model, zebrafish model and fungus C. elegans model in metabolite identification based on the structure and classification of synthetic cannabinoids.

Feasibility and clinical value of computed tomography-guided125I brachytherapy for pain palliation in patients with retroperitoneal lymph node metastases

Aims: The aim was to evaluate the feasibility and clinical value of computed tomography (CT)-guided125 I brachytherapy for pain palliation in patients with retroperitoneal lymph node metastases. Materials and Methods: A total of 23 patients with retroperitoneal lymph node metastases and those who had moderate-to-severe pain from January 2014 to December 2018 were enrolled in the study. The primary tumors included pancreatic (n = 12), gastric (n = 4), hepatocellular (n = 4), colorectal (n = 2), and esophageal carcinomas (n= 1). Patients were treated with CT-guided percutaneous125 I brachytherapy during the study. The Brief Pain Inventory-Short Form was used to record and compare pain intensity and interference by pain. Treatment-related complications were also evaluated according to the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer Late Radiation Morbidity Scoring Criteria. Statistical analysis was performed using SPSS software version 22.0 Results: The primary success rate of125 I seed implantation was 95.7% (22 of the 23 patients). As pain evolved, the patients achieved obvious pain palliation ratings for “worst pain” and “average pain” at 72 h and 4 weeks after brachytherapy, respectively, whereas “pain right now” at 12 weeks was significantly relieved after brachytherapy. No serious complications developed during the perioperative period. Conclusions: In the treatment of intractable carcinomatous pain in patients with retroperitoneal lymph node metastases, CT-guided125 I brachytherapy is a feasible and effective modality for pain palliation

Periodontal health knowledge and behavior among adolescents aged 12-15 years in Henan Province / 中国学校卫生

Objective@#To explore periodontal health knowledge and behavior among adolescents aged 12-15 years in Henan Province so as to provide scientific basis for oral health care.@*Methods@#According to the basic methods of the Fourth National Oral Health Survey, 3 786 school adolescents, aged 12-15 years, were selected from 12 middle schools in two district and two country in Henan by multi-stage stratified equal capacity random sampling. Oral examination and a questionnaire survey were performed among these students.@*Results@#The rate of periodontal health was 6.08%, the prevalence of gingival bleeding in these adolescents was 93.92%, its calculus existence was 90.99%. The rate of brushing teeth was 77.79%. However, the rate of brushing teeth twice or more per day was 17.56%, the rate of using fluoride toothpaste was 4.57%. The rate of periodontal health of adolescents who brushing teeth, using fluoride toothpaste were higher than adolescents who did not brush teeth, did not use fluoride toothpaste (χ2=10.81, 5.96, P<0.05). The recognition of oral care knowledge was 62.31%; except for "bacteria can cause dental caries", the awareness rate of other related periodontal diseases is more than 50.00%.@*Conclusion@#The status of periodontal health among adolescents aged 12-15 in Henan Province was not optimistic. Adolescents are lake of knowledge and health behavior towards periodontal health, The promotion of adolescents’s oral health education should be strengthened.

DKK1 expression in head and neck squamous cell carcinoma: analysis of its clinical value and regulatory factors / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the expression and regulation mechanism of Dickkopf-1 (DKK1) in head and neck squamous cell carcinoma (HNSCC) tissues. Methods: Based on the TCGA database, the relationship of DKK1 expression in HNSCC tissues and its methylation site with patients’prognosis was analyzed. GO and KEGG gene enrichment method were used to analyze the signaling pathways of DKK1 enrichment. STRING was used to analyze the interaction between DKK1 protein and other proteins. TargetScan was used to analyze the miRNAs that regulate the expression of DKK1, and the transcription factors of DKK1 were analyzed with the TRRUST website. Results: DKK1 gene was highly expressed in HNSCC tissues (P<0.01), and its expression level was significantly correlated with the HPV status, age, pathological grade, and clinical stage of patients (all P<0.05); the prognosis of HNSCC patients with high DKK1 expression was poorer than those with low DKK1 expression (P<0.01). There were 19 methylation sites in DKK1, 12 of which were significantly different between cancer tissues and normal tissues (P<0.05), and 11 sites were significantly related to the prognosis of HNSCC (P<0.05). In addition, miRNA, circRNA, lincRNA and transcription factors, etc. also participated in the regulation of DKK1. A total of 5 DKK1-related PPI networks that may involve in the occurrence, development, invasion and metastasis of HNSCC were obtained. Conclusion: DKK1 is highly expressed in HNSCC tissues and is a risk factor for poor prognosis of HNSCC patients. DKK1 plays an important role in the pathogenesis of HNSCC and is expected to become a potential target for HNSCC treatment.

Three-year effect evaluation of pit-and-fissure sealing of the first permanent molars in Henan provincial children / 中国学校卫生

Objective@#To examine the retention rate and caries prevention effect of fit-and-fissure sealants after 3 years of children oral disease comprehensive intervention program in Henan province.@*Methods@#For each of the five provincial regions attending the program in 2015, two classes from grade 2 in four primary schools were selected. Children in two schools were assigned into intervention group and the first permanent molars which were appriopriate for sealing were sealed with pit-and-fissure sealants. Children in other two schools were included in the control group and the first permanent molars were only examined for caries conditions. Three years later, retention rate of pit-and-fissure sealants and the dental caries prevalence was compared.@*Results@#Complete retention rate of sealants in the first permanent molars was 65.55% and the whole retention rate was 94.78%. The caries incidence and mean value of the first permanent molars in the intervention group children was 9.71%, (0.12±0.40) respectively whereas those in control group was 21.90%, (0.39±0.78) respectively. The differences between two groups were statistically significant(χ2/t=27.76, 6.91, P<0.01). The relatively effective rate of sealing for caries reduction was 63.43% and the actural effective rate was 6.51% and the pure yield rate of sealing was 12.19%. Higher sealant retention rate was associated caries prevalence(χ2=153.28, P<0.01).@*Conclusion@#Childhood oral disease intervention project in Henan Province shows positive effects. Sealing of the first permanent molars effectively prevent dental pit and fissure caries.

Discovery and study on potential effect of herbal pair of Uncariae Ramulus cum Uncis-Eucommiae Cortex on pregnancy hypertension based on network pharmacology and molecular docking / 中国中药杂志

This study aimed to explore the optimal indications and mechanism of Uncariae Ramulus cum Uncis(UR)-Eucommiae Cortex(EC) in lowering blood pressure based on network pharmacology and molecular docking. Chemical constituents were collected and screened by TCMSP database. Swiss Target Prediction platform was used to predict the related targets of the drug. OMIM, TCMIP and GeneCards databases were used to collect hypertension-related genes, and the intersections were taken to obtain potential targets for anti-hypertensive treatment of UR-EC. FunRich software was used to enrich the clinical phenotype and expression site of potential target of lowering blood pressure to analyze and predict the optimal indications of UR-EC. STRING database was used for KEGG pathway enrichment analysis, and Cytoscape 3.7.2 was used to construct the network of "composition-target-pathway". The key targets and their corresponding components in the network were analyzed and obtained, and then molecular docking was applied for preliminary verification. Twenty potential active components of UR and 24 potential active components of EC were respectively collected, and 92 anti-hypertensive potential targets of UR-EC were obtained. According to FunRich enrichment results, the optimal indication of UR-EC was pregnancy hypertension, which involved calcium signaling pathway, HIF-1 signaling pathway, neuroactive ligand receptor interaction, renin vascular tightening, VEGF signaling pathway, etc. In addition, AKT1, NOS2, ADRB2, F2, NOS3, SCN5 A, HTR2 A and JAK2 were considered as the key targets in the network. The molecular docking results showed that the screened potential active components had high binding activity with the key targets. This study preliminarily revealed that UR-EC may have therapeutic effects on pregnancy hypertension in terms of sedation, anti-hypertension, anti-inflammatory, anti-oxidation, improvement of vascular endothelial function and so on.

Cardiovascular events in hyperuricemia population and a cardiovascular benefit-risk assessment of urate-lowering therapies: a systematic review and meta-analysis / 中华医学杂志(英文版)

BACKGROUND@#Hyperuricemia and gout have become public health concerns; many important guidelines have recommended xanthine oxidase inhibitors (XOIs) as the first-line urate-lowering therapies (ULTs) to treat chronic gout with hyperuricemia. However, whether treating hyperuricemia and gout with ULTs modifies cardiovascular risks remains controversial. The aim of this study was to assess the incident risk of cardiovascular (CV) events (CVE) in hyperuricemia population, assess the cardiovascular benefit-risk of ULTs in hyperuricemia patients with or without gout in diverse cardiovascular risk sub-groups, and specify the safety of different ULTs.@*METHODS@#We searched PubMed, Embase, the Cochrane Library, Wanfang, Chongqing VIP (CQVIP, en.cqvip.com), and China National Knowledge Infrastructure Database for prospective cohort studies and randomized controlled trials (RCTs) in English and Chinese. Potential medications included XOIs, and uricosurics. RCTs were divided into sub-groups analysis based on blinding status and patients' history of CV diseases. Risk ratios (RRs) were calculated and were reported with corresponding 95% confidence intervals (CIs) by fixed-effects or random-effects model.@*RESULTS@#Seven prospective cohort studies and 17 RCT studies were included. The risks of both major adverse cardiovascular events (MACE) (RR = 1.72, 95% CI 1.28-2.33) and CVE (RR = 1.35, 95% CI 1.12-1.62) were higher in the hyperuricemia population than non-hyperuricemia one. In seven RCT studies where XOIs were compared with no-treatment or placebo, the results of five low CV risk studies showed that XOIs lowered the risks of both MACE (RR = 0.35, 95% CI 0.20-0.62) and CVE (RR = 0.61, 95% CI 0.44-0.85); whereas two high CV risk studies showed that XOIs lowered the risk of CVE (RR = 0.69, 95% CI 0.54-0.88) rather than MACE (RR = 0.62, 95% CI 0.29-1.35). In nine RCT studies where the cardiovascular safety between febuxostat and allopurinol were compared, no statistical difference was found in the risk of MACE or CVE.@*CONCLUSIONS@#The hyperuricemia population does have a higher incidence of CVE, and the results suggested that XOIs might reduce the incidence of MACE and total CVE. In addition, from the perspective of cardiovascular safety, febuxostat equaled allopurinol in our meta-analysis.

Deciduous dental caries among children aged 3-5 years in Henan Province and its influencing factors / 中国学校卫生

Objective@#To investigate the epidemiology and associated factors of deciduous dental caries in 3-5 years old children in Henan Province, and to provide a epidemiological reference for prevention work of deciduous dental caries among children.@*Methods@#According to the basic methods of the Fourth National Oral Health Survey, 12 kindergartens were randomly selected from 2 counties and 2 districts in Henan Province and a total of 1 296 children aged 3-5 years old were randomly selected with stratified multi-stage random sampling method. The deciduous teeth of these children were examined for caries and their guardians were surveyed with questionnaires regarding oral health behavior, knowledge and attitude.@*Results@#Deciduous dental caries occurred in 755 children, accounting for 58.3% and the average decay-missing-filled teeth (dmft) was 2.83. The differences of prevalence of dental caries of boys and girls (60.5%, 56.0%) and mean dmft index (2.88, 2.78) were of no statistical significance(χ2/F=2.64, 0.22, P>0.05). The prevalence of dental caries and mean dmft index for 3, 4 and 5-year-old group was 46.7%, 2.02 and 60.9%, 2.85 and 67.5%, 3.64, respectively. There was statistical significance among three age groups(χ2/F=40.00, 21.68, P<0.01). No significant difference was found in the dental caries prevalence and mean dmft index between urban (58.9%, 2.87) and rural area (57.7%, 2.79)(χ2/F=0.20, 0.16, P>0.05). Multivariate logistic regression showed that age, feeding pattern within the first six months after birth, dental care experience and self-perceived oral health status associated with deciduous caries in 3-5 year-old children in Henan Province(P<0.05).@*Conclusion@#Dental caries prevalence is highly prevalent among preschool children in Henan Province. Effective caries precaution measures should be taken to improve oral health level of these children.

Identification of New Designer Benzodiazepine Diclazepam in Drug Facilitated Sexual Assault / 法医学杂志

OBJECTIVES@#To identify the new designer drugs which are totally unknown and not in the routine testing list by the technologies such as high-resolution mass spectrometry in drug facilitated sexual assault, in order to solve the problem in actual cases.@*METHODS@#The milky fluid from an actual case was extracted and analyzed using LC-QE, ¹H-NMR and GC-MS, respectively. The accurate masses and cluster ions isotope patterns of unknown compound were obtained by LC-QE. The molecular formula was confirmed as C₁₆H₁₂C₂N₂O based on the protons number of ¹H-NMR. The isomers diclazepam and 4-chlorodiazepam were separated and detected with GC-MS.@*RESULTS@#The new designer benzodiazepine as diclazepam in the milky fluid was identified. The results provided direct evidence for the investigation and qualitative analysis of such cases.@*CONCLUSIONS@#The combined application of various methods, including LC-QE, ¹H-NMR and GC-MS, can be used to detect unknown new psychoactive substances.

Role of leukocyte immunoglobulin like receptor subfamily B in tumorigenesis and development / 中国肿瘤生物治疗杂志

@#白细胞免疫球蛋白样受体亚家族B（leukocyte immunoglobulin-like receptor subfamily B，LILRB）在骨髓细胞、造血干 细胞、神经细胞等多种机体细胞广泛表达。有研究发现，LILRB受体可以与多种配体结合并具有多种生物学功能，包括调节炎症 反应、免疫耐受、细胞分化和神经系统的可塑性等。近年来研究发现，LILRB在多种实体瘤和血液系统肿瘤表达增高并与患者预 后显著相关，同时LILRB与免疫抑制、肿瘤细胞生长和自我更新直接相关，具有肿瘤支持因子和免疫检查点分子的双重作用。此 外，肿瘤细胞表达的LILRB与肿瘤微环境中的免疫细胞相互作用，调节机体对肿瘤的免疫反应，敲除小鼠的LILRB同源基因后， 小鼠的正常造血功能未受到明显影响。上述研究结果提示，LILRBs可能是肿瘤治疗的理想靶点。本文就LILRB与实体瘤和血 液系统肿瘤的发生机制、LILRB在肿瘤细胞中的信号转导方式、LILRB与肿瘤的免疫治疗及需要解决的问题进行阐述，以期为后 续的深入研究提供参考。

Effects of CXCL4 on human dermal microvascular endothelial cells and secretion of vasomotion factors / 中国免疫学杂志

Objective:To identify the proliferation effect and angiogenic ability of CXCL4 on human dermal microvascular endothelial cells(HDMECs),and to explore the secretion of vasomotion factors.Methods:HDMECs were treated with gradient concentration to test the proliferation of HDMECs.CCK-8 was used to explicated the proliferation of HDMECs.The effect of CXCL4 on angiogenic ability of HDMECs was determined by tube formation assay.The mRNA levels of endothelin-1(ET-1),Fli-1,AngiotensinⅡtype 1 receptor(AT1R) and endothelin-1 type A receptor (ETAR) were detected by real-time quantitative polymerase chain reaction (Real-time PCR).Results:The specific receptor of CXCL4 was expressed on HDMECs.CXCL4 could inhibit the proliferation of HDMECs and the number of tube formation in a dose-depend manner.After CXCL4 intervention,the relative amplification multiples of ET-1,AT1R were significantly increased(P<0.05),Fli-1 was decreased(P<0.05),and ETAR had no change as compared with the control group.Furthermore,CXCL4 antagonist could reverse the effects of CXCL4 on HDMECs.Conclusion:CXCL4 inhibit the proliferation and angiogenesis of HDMECs and induce the secretion of ET-1 and AT1R,reduce the secretion of Fli-1 in a dose-dependent manner.

Interactions between induced T regulatory cells and B cells in inflammatory milieu in mice with collagen-induced arthritis / 中国免疫学杂志

Objective:To investigate the interactions between induced T regulatory cells ( iTregs ) and B cells in the inflammatory milieu in mice with collagen-induced arthritis(CIA). Methods: CD19+ cells were isolated from the spleen cells of normal DBA1/J(N-B) mice and CIA mice(CIA-B) on the 35th day after the first immunization with established arthritis. These B cells were used as antigen-presenting cells to observe their effects on the induction of Tregs. Tregs were induced with the classic method and co-cultured with CIA-B cells. CIA-B cell effects on iTreg proliferation and the expression of CTLA-4 on iTregs were explored. iTregs′ influence on the expressions of co-stimulators(CD80,CD86) and MHCⅡon B cells was studied and its mechanism was determined by the Transwell experiments. Results: CIA-B could induce more Treg production and proliferation. CIA-B could also promote the CTLA-4 expression on iTreg cell surface which worked through a cell-contact pathway, while iTregs could increase the expressions of co-stimulators(CD80,CD86) and MHCⅡby the same way. Conclusion: iTregs could show their immune suppressive function through the interactions with CIA-B cells in the inflammatory milieu in mice with CIA. These interactions work by a cell-contact pathway.

Redox regulation of c-JNK signaling pathway on myocardial potassium channel reconstruction in diabetic rats / 解放军医学杂志

Objective To investigate the role of c-Jun NH2-terminal kinase (c-JNK) signaling pathway on voltage-gated potassium channel (Kv) remodeling in left ventricular myocytes of diabetic rats,and explore the intrinsic regulatory mechanism.Methods Forty-five SD rats were randomly divided into DM group (n=25,modeling with streptozotocin induction) and control group (n=20,fed with normal diet).Transient outward potassium current (Ito) of rats' ventricular myocytes in DM group and control group was recorded by whole-cell patch-clamp method.The c-Jun activity was detected using a non-radioactive JNK kinase assay kit (Cell Signaling Technology).JNK inhibitor SP600125 was used to incubate the cardiomyocytes of diabetes rats in vitro,and then the changes of I,o in cardiomyocytes were observed.Thioredoxin reductase (TrxR) inhibitor--auranofin (AF) was used to treat the rats' cardiomyocytes incubated with SP600125,and then the changes of Ito in cardiomyocytes were observed.The content of Kv4.2 was tested using anti-Kv4.2 antibody,and the results were analyzed using a UVP bioimaging system.Results The JNK activity in DM group rose more than 1 times compared with control group,while the density of Ito decreased significantly (Control:30.2 ± 3.3pA/pF,n=16;DM:15.3 ± 2.1pA/pF,n=17;P＜0.05).The ventricular myocytes of DM rats were treated with SP600125 (10μmol/Lol/L) for 4 hours,then the Ito density increased to control group level (DM+SP600125:32.3 ± 3.7pA/pF,n=18;Control:30.2 ± 3.3pA/pF,n=16;P＜0.05).There was no significant difference in the maximum Ito density between the treated with SP600125 (Control+SP600125:31.6 ± 3.4pA/pF,n=18) and untreated control groups.The Ito density in DM myocardial cells significantly increased after treatment with the membrane permeable protein inhibitor JNKI-1 (10μmol/L),and no changes were found in control group after the same treatment.The augmentation effect of SP600125 on Ito current in DM myocytes was significantly inhibited by TrxR inhibitor auranofin (lμmol/L) (DM+SP600125+AF:15.7 ± 3.3pA/pF,n=15),while AF did not change the Ito density in control group.The expression of Kv4.2 protein was significantly increased in DM rats after administration of SP600125,which was consistent with the changes of Ito current observed in the myocardium of DM rats,although not fully restored to the level of control group myocardium.JNK inhibitor did not markedly alter the expression of Kv4.2 protein in control group myocardium.Conclusions Kv channel remodeling in DM rat's myocardium is redox-regulated,and the Ito remodeling might be assisted with the persistent activation of c-JNK signaling pathway.It has showed that c-JNK activity is significantly increased in DM rat heart and the current density of Kv channels is reduced.The inhibition of JNK signaling pathway can markedly improve Kv channel reconstruction and the process may be regulated by thioredoxin system.

Identification of Methamphetamine Abuse and Selegiline Use: Chiral Analysis of Methamphetamine and Amphetamine in Urine / 法医学杂志

Objective To study the content variation of selegiline and its metabolites in urine, and based on actual cases, to explore the feasibility for the identification of methamphetamine abuse and selegiline use by chiral analysis. Methods The urine samples were tested by chiral separation and LC-MS/MS method using CHIROBIOTICTM V2 chiral liquid chromatography column. The chiral analysis of metham-phetamine and amphetamine were performed on the urine samples from volunteers of selegiline use and drug addicts whom suspected taking selegiline. Results After 5 mg oral administration, the positive test time of selegiline in urine was less than 7 h. The mass concentrations of R(-)-methamphetamine and R(-)-amphetamine in urine peaked at 7 h which were 0.86μg/mL and 0.18μg/mL and couldn't be de-tected after 80 h and 168 h, respectively. The sources of methamphetamine and amphetamine in the urine from the drug addicts whom suspected taking selegiline were analysed successfully by present method. Conclusion The chiral analysis of methamphetamine and amphetamine, and the determination of selegi-line's metabolites can be used to distinguish methamphetamine abuse from selegiline use.

Prevalence of mitochondrial DNA mutations in sporadic patients with nonsyndromic sensorineural hearing loss / Prevalência de mutações no DNA mitocondrial em pacientes esporádicos com deficiência auditiva sensorioneural não sindrômica

ABSTRACT INTRODUCTION: Several mitochondrial DNA mutations have been reported to be associated with nonsyndromic hearing loss in several families. However, little is known about the prevalence of these mutations in sporadic patients with nonsyndromic sensorineural hearing loss. OBJECTIVE: The purpose of our study was to investigate the incidence of these mitochondrial DNA mutations in such population. METHODS: A total of 178 sporadic patients with nonsyndromic sensorineural hearing loss were enrolled in this study. Genomic DNA was extracted from the peripheral blood sample. We employed the SNaPshot(r) sequencing method to detect five mitochondrial DNA mutations, including A1555G and A827G in 12S rRNA gene and A7445G, 7472insC, and T7511C in tRNASerUCN gene. Meanwhile, we used polymerase chain reaction and sequenced the products to screen GJB2 gene mutations in patients carrying mitochondrial DNA mutations. RESULTS: We failed to detect the presence of A1555G mutation in 12S rRNA gene, and of A7445G, 7472insC, T7511C mutations in tRNASerUCN gene in our population. However, we found that 6 patients (3.37%) were carriers of a homozygous A827G mutation and one of them also carried homozygous GJB2 235delC mutation. CONCLUSION: Our findings in the present study indicate that even in sporadic patients with nonsyndromic sensorineural hearing loss, mitochondrial DNA mutations might also contribute to the clinical phenotype.

Resumo Introdução: Diversas mutações do DNA mitocondrial tem sido descritas, em diferentes famílias, associadas à deficiência auditiva não sindrômica. No entanto, pouco se sabe sobrea prevalência dessas mutações em pacientes esporádicos com deficiência auditiva sensorioneural não sindrômica. Objetivo: A finalidade do nosso estudo foi investigar a incidência dessas mutações no DNA mitocondrial nessa população. Método: No total, 178 pacientes esporádicos com deficiência auditiva sensorioneural não sindrômica foram recrutados para participação no estudo. O DNA genômico foi extraído de amostra, de sangue periférico. Utilizamos o método de sequenciamento SNaPshot(r) para detecção de cinco mutações do DNA mitocondrial: A1555G e A827G no gene 12S rRNA e A7445G, 7472insCe T7511C no gene tRNASerUCN. Paralelamente, utilizamos a reação de polimerase em cadeia e sequenciamos os produtos para triagem das mutações no gene GJB2 nos pacientes portadores de mutações no DNA mitocondrial. Resultados: Em nossa população, não conseguimos detectar a presença da mutação A1555G no gene 12S rRNA e nem as mutações A7445G, 7472insC e T7511C no gene tRNASerUCN. Entretanto, constatamos que seis pacientes (3,37%) eram portadores da mutação homozigota A827G; e um deles também portava a mutação homozigota GJB2 235delC. Conclusão: Nossos achados no presente estudo indicam que, mesmo em pacientes esporádicos com deficiência auditiva sensorioneural não sindrômica, as mutações do DNA mitocondrial também podem contribuir para o fenótipo clínico.

Comparison of Drug Release in vitro of Diclofenac Sodium Sustained-release Capsules and Sustained-release Tablets from Different Manufacturers / 中国药师

Objective: To establish an HPLC method for the determination of diclofenac sodium sustained-release preparations, and compare the in vitro dissolution of diclofenac sodium sustained-release capsules and tablets from different manufacturers to provide reference for the clinical reasonable medication. Methods: According to the first dissolution method described in Chinese Pharmaco-poeia (2010 edition),the dissolution of each sample was determined by HPLC and the cumulative release percentage was calculated. The dissolution parameters were fitted by Weibull-equation and the results were analyzed. Results:The HPLC method for the determi-nation of diclofenac sodium release was accurate with good reproducibility, and the dissolution parameters of different manufacturers had remarkable difference. Conclusion:The in vitro dissolution of diclofenac sodium sustained-release capsules and tablets from different manufacturers is various, which should be paid attention in the clinical use.