Carcinoma, Squamous Cell ; pathology ; Child ; Cysts ; pathology ; Diagnosis, Differential ; Epithelium ; pathology ; Female ; Follow-Up Studies ; Humans ; Keratin-19 ; metabolism ; Keratin-5 ; metabolism ; Keratin-6 ; metabolism ; Keratin-7 ; metabolism ; Keratins ; metabolism ; Neoplasm Recurrence, Local ; surgery ; Reoperation ; Submandibular Gland ; surgery ; Submandibular Gland Neoplasms ; metabolism ; pathology ; surgery ; Transcription Factors ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To evaluate the molecular mechanism of 11 beta-hydroxysteroid dehydrogenase type 1 (11 β-HSD1)in tumor necrosis factor-α (TNF-α) induced insulin resistance.Methods The optimal concentration and incubation time of TNF-α for treating HepG2 cells were selected.Real-time polymerase chain reaction(PCR)and Western blot analysis were used to explore the effect of TNFα on the mRNA and protein expression levels of 11β-HSD1,glucocorticoid receptor (GR),phosphoenolpyruvate carboxylase(PEPCK)and glucose 6-phosphatase(G6Pase).Glucose(GO) Assay Kit was used to determine the level of gluconeogenesis.Liver/Muscle glycogen assay kit,PAS/Glycogen Stain kit were used to detect glycogen synthesis.The expression of AKT/GSK pathway was analyzed by Western blot analysis.In addition,HepG2 cells were pre-treated with BVT2733,a specific inhibitor of 11β-HSD1,before exposure to TNFα The expression of AKT/GSK pathway,the level of gluconeogenesis and glycogen synthesis were analyzed.Results The expression of 11β-HSD1 was increased along with the increasing concentration of TNF-α,showing a dose-dependent effect.When HepG2 cells treated with the optimal dose of TNF-α 10 μg/L for 24 hours,the levels of GR,gluconeogenesis and gluconeogenesis-related genes PEPCK and G6Pase were increased,while conversely,the levels of phosphorylation of AKT and GSK,and glycogen synthesis were decreased.However,all the above effects induced by TNF-α in HepG2 cells were greatly reversed by BVT2733 pre treatment.Conclusions 11 β-HSD1 participates in TNF-α-induced insulin resistance in HepG2 cells,and BVT2733 could significantly improve insulin sensitivity.

Objective To investigate gene polymorphisms of drug targets and mutations associated with drug resistance in Pneumocystis jiroveci (P.jiroveci) isolates.Methods Among 148 samples isolated from human immunodeficiency virus (HIV)infected patients with pneumonia in Guangdong,mitochondrid larg subunit rRNA (mtLSUrRNA) gene was amplified from 51 samples.Dihydropteroate synthase (DHPS),dihydrofolate reductase (DHFR) and Cytochrome b (CYB) genes of P.jiroveci were detected by gene sequencing,and compared with the reference sequences in GenBank to evaluate gene polymorphisms.Results P.jirovecii DHPS,DHFR and CYB genes were all successfully amplified from 51 samples.For DHPS gene,48 (94.1％) were wild-type and 3 (5.9％) had gene mutation associated with drug resistance.For DHFR gene,30 were wild-type,and 21 had a synonymous mutation at position 312,and 1 nonsynonymous mutation at position 188.There were no mutations associated with drug resistance.For CYB gene,polymorphisms of were detected at 5 sites,4 of which were synonymous mutations,1 was non-synonymous mutation.No mutation associated with drug resistance was found.Based on the gene polymorphism of CYB6,the strains can be classified into 6 genotypes,and 2 were first detected,including 25 CYB1,13 CYB2,2 CYB5,4 CYB8,as well as newly detected 4 CYB10 and 3 CYB11 strains.Conclusions The mutations associated with drug resistance in P.jiroveci isolates in Guangdong remain uncommon.CYB gene shows gene polymorphisms and can be selected as one of targeted genes for multilocus sequence typing.

To improve the efficiency of halophilic actinobacteria screening and carry out the rapid selection of targeted strains, we tested 34 strains of Streptomonospora by PCR-single strand conformation polymorphism analysis (PCR-SSCP) based on genus-specific primers for the PCR identification. This approach employs PCR with two pairs of primers located in the 16S rRNA sequence flanking two variable region, then build clustering tree according as SSCP data. Synchronously, we sequenced all the 16S rRNA partial sequences for these strains to verify them. The results showed that the PCR-SSCP analysis was an efficient, easy-to-handle and economic method for rapid selection of halophilic actinobacteria resources.

Objective To investigate the post-traumatic stress disorder (PTSD) and its influence factors among people living with HIV /AIDS (PLWHA) ,and provide evidence for developing relevant interventions and treatment methods .Methods Anony-mously completed questionnaires ,including self-designed questionnaire and the post-traumatic checklist civilian version (PCL-C) , were received from PLWHA in Suizhou from December 2014 to March 2015 .Descriptive analysis and multivariate logistic regres-sion analysis were used to examine factors associated with PTSD .Results Among 243 qualified PLWHA ,106 of them were positive with PTSD (43 .62% ) ,and the prevalence of PTSD among male and female were 37 .88% and 50 .45% ,respectively .Among the three characteristic symptoms ,repeated flashback was the highest (78 .60% ) ,then hyper-vigilance (64 .20% ) ,and the positive rate of avoidance/numbness was the lowest (57 .20% ) .Multivariate logistic regression analysis showed that female was more suscepti-ble to PTSD than male (OR = 1 .960 ,95% CI :1 .025 - 3 .747) ,discrimination or unfair treatment would increase the risk of PTSD (OR= 2 .967 ,95% CI :1 .498 - 5 .876) ,and the frequency of fear/tension was positive correlated with PTSD (occasionally OR =2 .074 ,95% CI :1 .008 - 4 .265 ;always OR = 6 .690 ,95% CI :2 .708 - 16 .531) .Conclusion PLWHA had a high level of PTSD ,and fe-male PLWHA and those who felt fear/tension or experienced discrimination/unfair treatment were more likely to suffer from PTSD .

Objective: To determine total phenylethanoid glycoside and acteoside in Plantago Herba to provide reference for evaluating the quality of medicinal materials.Methods: With acteoside as the control sample, a UV visible spectrophotometric method was used to determine total phenylethanoid glycosides in Plantago Herba.An HPLC method was applied to determine acteoside in Plantago Herba , and the conditions were as follows: an ODS2 C 18 (150 mm× 4.6 mm ,5 μm) chromatographic column was used with acetonitrile-0.1% formic acid (13∶87) as the mobile phase at a flow rate of 1.0 ml·min-1 , the detection wavelength was 332nm, the column temperature was 30℃, and the sample volume was 10 μl.Results: The reference solution and the sample solution had the maximum absorption at 332 nm, and the linear relationship was good within the range of 0.003 1-0.155 0 mg·ml-1 (r=0.999 5).The content of total benzene alcohol glycosides in 3 batches of samples was 2.73% , 2.61% and 2.84% , respectively;acteoside over the range of 0.000 6-0.155 0 mg·ml-1 (r=0.999 1) showed a good linear relationship with peak area,the sample recovery was 98.5% and the RSD was 1.6% (n =6), and the acteoside content in 3 batches of samples respectively was 0.54% , 0.51% and 0.56%.Conclusion: The method is simple, accurate and reproducible, and can be used for the determination of total phenylethanoid glycosides and acteoside in Plantago Herba.

Objective:To investigate the expression of serum human phosphatidylethanolamine-binding protein 4 (hPEBP4) in patients with multiple myeloma (MM) and its clinical significance.Methods:A total of 59 symptomatic MM patients admitted to West Branch of Beijing Chaoyang Hospital from September 2016 to September 2018 were selected as the research objects. According to the CRAB symptoms [elevated serum calcium (C), kidney injury (R), anemia (A), bone lesions (B)], all patients were divided into 2 groups, including the active group of 44 patients with CRAB symptoms, and the response group of 15 patients who achieved at least partial remission after chemotherapy and symptom relief of CRAB. According to the degree of bone lesions (BL), 30 patients with severe bone-related events were grouped as the severe bone lesions (SBL) group, and 14 patients were grouped as the non-severe bone lesions (NSBL) group. According to the revised international prognostic staging system (R-ISS), patients in the active group were divided into three subgroups: stage Ⅰ, stage Ⅱ, and stage Ⅲ, including 26, 11 and 7 patients, respectively. A total of 15 healthy examination people whose gender and age matched those of the patients were treated as the healthy control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of hPEBP4, tumor necrosis factor ligand superfamily member 14 (LIGHT/TNFSF14) and activin A of patients in different groups. Pearson was used to analyze the relationship of the expressions of multiple factors in the active group. The optimal cut-off value of multiple factors diagnosing MM was determined by using receiver operating characteristic (ROC) curve, and according to the cut-off value, the differences in overall survival (OS) of patients with different stratification were compared.Results:In the active group, the respond group, the healthy control group, the level of hPEBP4 was (1.48±0.64) μg/L, (1.49±0.75) μg/L, (0.31±0.10) μg/L, respectively; the level of LIGHT/TNFSF14 was (169±112) ng/L, (256±132) ng/L, (44±27) ng/L,respectively; the level of activin A was (383±266) ng/L, (223±79) ng/L, (234±85) ng/L, respectively; and the differences were statistically significant (all P<0.05). In the active group, the level of hPEBP4 was (1.06±0.60) μg/L, (1.15±0.50) μg/L, (1.73±0.68) μg/L, respectively in patients with stage R-ISSⅠ, R-ISSⅡ and R-ISS Ⅲ, and the difference was statistically significant ( F=3.287, P=0.032). The level of activin A was (219±55) ng/L, (247±117) ng/L, (450±215) ng/L, respectively among patients in stage R-ISSⅠ, R-ISSⅡ, R-ISS Ⅲ, and the level of activin A in stage R-ISS Ⅲ was higher than that in stage R-ISSⅠand R-ISSⅡ (all P < 0.05). The levels of LIGHT/TNFSF14 and activin A of SBL patients were higher than those of NSBL patients [(174±101) ng/L vs. (98±53) ng/L; (467±238) ng/L vs. (189±71) ng/L, all P < 0.05]. The level of hPEBP4 was positively correlated with the levels of M protein ( r=0.694, P < 0.01) and activin A ( r=0.252, P < 0.01) of IgG patients in the active group. ROC curve analysis showed that the optimal cut-off value of hPEBP4, LIGHT/TNFSF14, activin A diagnosing MM was 1.04 μg/L, 97.0 μg/L, 156.2 ng/L. The median overall survival (OS) time of patients with hPEBP4 >1.04 μg/L and hPEBP4 ≤ 1.04 μg/L was 57 months (95% CI 22-92 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05); while the median OS time of patients with activin A ≥ 156.2 ng/L and activin A < 156.2 ng/L was 61 months (95% CI 24-98 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05). Conclusions:High expression level of hPEBP4 is related with the progression of MM. It is positively related with the level of M protein and negatively with the OS of MM patients. It is suggested that hPEBP4 may be used as an important marker to judge disease progression and tumor burden in MM. LIGHT/TNFSF14 and activin A cooperate with hPEBP4 to participate in the pathological processes of tumor microenvironment of MM.

Objective To establish a method for determination of phenylethanoid glycoside and acteoside in Plantago Herba. Methods UV-visible spectrophotometric method was used for the determination of the content of phenylethanoid glycosides compounds in Plantago Herba. HPLC method was used for the determination of acteoside in Plantago Herba. Chromatographic column with C18 ODS2 (4.6 mm × 150 mm, 5 μm) was used. Acetonitrile-0.1%formic acid (13:87) was as mobile phase; the flow rate was 1 mL/min; the detection wavelength was 332 nm; the column temperature was 30 ℃; the sample volume was 10 μL. Results The contents of phenylethanoid glycoside in Plantago Herba from different producing areas were among 1.03%–3.47%. Acteoside with peak area over the 0.0062–1.55 mg range showed a good linear relationship; the sample recovery rate was 98.9%, and the RSD was 1.6%. The contents of acteoside in Plantago Herba from different producing areas was among 0.18%–0.56%. Conclusion The method is simple, stable and reproducible, which can be used for the determination of phenylethanoid glycoside and acteoside in Plantago Herba from different producing areas and provide experimental basis for quality control of Plantago Herba.

Objective: To observe the characteristics of dynamic changes of lysophosphatidic acid (LPA) levels in cerebrospinal fluid (CSF) in patients with subarachnoid hemorrhage (SAH) and its relationship with cerebral vasospasm (CVS) and to explore the pathogenesis of CVS. Methods: Sixty-seven patients with SAH diagnozed by clinical and accessory examinations were selected. The LPA levels in CSF were measured at 24 hours, day 7,14, and 28 respectively after the onset of symptoms,and they were compared with a control group. The correlation between LPA levels and CVS on the time course was also observed at the same time. Results: Of the 67 patients with SAH, a total of 29 patients (43.3%) occurred CVS, the average time of occurrence was 6. 6 days. There was no significant difference between the LPA levels in CSF in patients with SAH and the control group at 24 hours after the onset of symptoms; they were significantly higher than the control group at day 7 (P ＜0. 001); they were significantly higher than the control group at day 14 (P ＜ 0. 001), but they were significantly lower than those at day 7 (P ＜ 0. 01); they decreased to baseline at day 28, and there was significant difference compared with the control group. There was no significant difference between the LPA levels in the CVS group and those in the non-CVS group at 24 hours, they were significantly higher than those in the non-CVS group at day 7 (P ＜0. 001), they were still significantly higher than those in the non-CVS group at day 14 (P ＜0. 01); and there was no significant difference between the 2 groups at day 28. Conclusions: The LPA levels in CSF in patients with SAH increased significantly from day 7 to day 14 after the onset of symptoms, and they had obvious association with CVS on the time course. The detection of the LPA levels in CSF may have important significance in predicting the occurrence of CVS.

Objective To investigate the epidemiological characteristics of dengue fever in Shenz-hen city in 2014 and to analyze the evolutional characteristics of the epidemic dengue virus(DENV)strains in order to provide scientific guidelines for the prevention and control of dengue fever. Methods Descrip-tive epidemiological analysis was used to analyze the prevalence of dengue fever in Shenzhen city in 2014. Immunochromatography and real-time PCR were performed to detect the specific antibodies(IgM and IgG) and DENV nucleic acids in serum samples collected from suspected cases of dengue fever. Serum samples collected from the patients at early stage of dengue fever were used to infect the C6 / 36 cell line for further isolation of DENV strains. The types of isolated DENV strains were determined by using real-time PCR. E genes of the isolated DENV strains were amplified by RT-PCR and then sequenced. DNAStar and Clustslx (1. 83)softwares were used to analyze the homology between DENV strains isolated in Shenzhen and other areas. A phylogenetic tree based on the sequences of E genes of Shenzhen strains and other sequences of DENV reference strains downloaded from GenBank was constructed for further analysis. Results A total of 454 cases of dengue fever were reported in Shenzhen in 2014 with a male to female ratio of 1. 43 ∶ 1. Local patients accounted for 76. 21% and the rest 23. 79% were imported cases mainly from Southeast Asian and surrounding cities. There were 441 cases reported from September to November,accounting for 97. 14% of all reported cases. Most of the infected subjects were aged 20 to 50,accounting for 76. 73% . Of the 270 samples positive for DENV nucleic acids,strains of DENV-1,DENV-2,DENV-3 and DENV-4 accounted for 87. 41% ,8. 89% ,0. 37% and 2. 22% ,respectively. The phylogenetic tree analysis revealed that the DENV-1 strains belonged to two genotypes,which were genotypeⅠ and genotype Ⅴ. The DENV strains of genotypeⅠ were highly similar to the epidemic strain isolated in Shenzhen in 2010 and the genotype Ⅴstrains were first reported in Shenzhen. The homology analysis of the nucleotides of E genes showed that mi-nor differences in the nucleotide sequences were found between DENV-2 strains. All of the DENV-2 strains belonged to the genotype Ⅳ as indicated by the phylogenic tree. Conclusion There were 454 cases of den-gue fever(including both local and imported cases)reported in Shenzhen city in 2014,reaching an all-time high. DENV-1 was the predominant pathogen in combination with an increased infection of DENV-2. This study indicated that the prevalent DENV strains might be imported from Southeast countries and neighboring cities. Further researches should be conducted to analyze whether dengue fever is endemic in Shenzhen City.