Objective To explore the characteristics of bone mineral density (BMD) and the incidence of osteoporosis in partial androgen deficiency in aging male (PADAM) patients.Methods Fifty-six PADAM patients (PADAM group) and 51 healthy persons (control group) were selected according to their age and body mass index (BMI),and measured their BMD in the second to fourth lumber and the neck of femur with the dual-energy C X-ray BMD measuring instrument.MES-01S20 muscle function analyzer was used to determinate the distribution and mechanical properties of muscle.The biochemical and bone metabolic markers and sexual hormones were collected and observed by multivariate stepwise regression analysis.Results Compared with control group,BMD significantly decreased in the Ward triangle,femoral neck and big rotor (P ＜ 0.01 ) and no significant change in lumbar in PADAM group (P ＞ 0.05 ),and the fracture strength of femoral neck (FS) and the lower-limb muscular strength (MS) also significantly decreased (P ＜ 0.01 ).The incidence of osteopenia were 48.2% (27/56) and 35.3% ( 18/51 ),osteoporosis were 30.4% (17/56) and 21.6% ( 11/51 ) respectively in PADAM group and control group.There was significant difference between two groups (P＜0.05).The BMD was positively correlated to BMI at the first to fourth lumber and negatively correlated to age,positively correlated to BMI and serum level of andrusol at the proximal left femur in the patients with PADAM.Conclusion BMD and the resistance to fracture aresignificantly lower in PADAM patients,and aging lower BMI and androgen deficiency are the risk factors of low BMD in PADAM patients.There is the potential of osteoporotic fracture risk,and it is important to strengthening the prevention and treatment of fractures in elderly PADAM patients.

In 2017, following many thorough discussions, considering Chinese actual situation, more than 20 distinguished pancreatic surgeons brought about an update of the previous 2010 Chinese experts′ consensus on the prevention and treatment of common complications after pancreatic surgery. Referred to the latest update of the postoperative pancreatic fistula consensus statement by the International Study Group of Pancreatic Surgery, the postoperative pancreatic fistula system of 2017 version Chinese consensus divided pancreatic fistula into pure fistula and mixed fistula based on whether other digestive fluid is mixed or not. The new version also presents key points of pancreatic fistula prevention and surgical strategy. In the paper, the authors analyzed the necessity, essentials and controversy of the update.

OBJECTIVE: To investigate the expression of EMS1 and DcR3 in laryngeal carcinoma and analyze the relation of EMS1 and DcR3. METHOD: The expression of EMS1 and DcR3 protein in 41 laryngeal carcinoma fresh samples and 41 para-carcinoma tissues (to cutting margin > 0.5 cm) were measured by flow cytometry, and 15 normal laryngeal mucosa samples were also studied as controls. RESULT: (1) The quantitative and qualitative expression of EMS1 and DcR3 protein in laryngeal carcinoma tissues was obviously higher than those in para-carcinoma and in normal laryngeal mucosa tissues respectively (P < 0.05). There was no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues. (2) In laryngeal carcinoma, the expression of EMS1 and DcR3 protein was independent of patients' clinical classification, tumor size, smoking history, patients' age and sex but associated with tumor metastasis, pathological grade and clinical stage. (3) In laryngeal carcinoma, the expression of EMS1 was positively correlated with that of DcR3. CONCLUSION EMS1 was positively related to DcR3, which might play an important role in the carcinogenesis and development of laryngeal carcinoma by synergic effect.
Adult ; Aged ; Cortactin ; metabolism ; Female ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

Objective To explore the therapeutic effect and clinical significance of pelvic floor rehabilitation technique in female with myofascial chronic pelvic pain (MCPP) by detecting the pressure pain thresholds (PPTs). Methods One hundred healthy female (control group) and 324 female MCPP patients (observation group) from January 2009 to December 2016 were selected. Automatic body surface and vaginal pressure pain detector was applied to detect two groups′PPTs of the 34 spots. The difference of PPTs at each spot was analyzed in two groups. In addition, 51 patients with moderate and severe MCPP were selected to record the changes of PPTs and pain scores before and after the treatment of pelvic floor rehabilitation technique. Results The average PPTs of the abdomen, vulva, pelvic floor and vaginal front and back fornix, bilateral adnexa uteri and sacrouterine ligament in the observation group were significantly lower than those in the control group (P<0.01 or<0.05). The average PPTs of the abdomen, vulva, pelvic floor muscles and vaginal front and back fornix, bilateral adnexa uteri and sacrouterine ligament of 51 MCPP patients after treatment were significantly higher than those before treatment (P<0.01). After treatment, PPTs and pain scores of the pelvic floor muscles, bilateral adnexa uteri, bilateral sacrouterine ligament, bilateral sacral spine ligament and vaginal front and back fornix were negatively correlated (r =- 0.78 to- 0.19, P = 0.01 to 0.04); there was a negative correlation between the PPTs and pain scores of the left and right latissimus dorsi (r=-0.28, P=0.04;r=-0.32, P=0.02). The complete remission rate with the pelvic floor rehabilitation technique in 51 patients with MCPP was 9.8％(5/51), the significant remission rate was 90.2％(46/51), and the total remission rate was 100.0％ (51/51). Conclusions Compared with the normal healthy ones, female with MCPP has lower PPTs in the abdomen, perineum, vagina and pelvic floor. The effect of pelvic floor rehabilitation technique on MCPP is well, which can increase patients′PPTs to reduce pain scores. It is a worthwhile method to treat these diseases.

Objective To study the effect of amino acid site variation of HIV-1 Nef protein on its inhibition of neuronal autophagy and explore the mechanism of Nef protein-induced central nervous system injury.Methods HIV-1 nef genes were amplified and cloned from the temporal cortex (TC) of the central nervous system in 1 case of HIV-associated dementia (HAD) H and 1 case of non-HAD AIDS patient N.The amino acid sequences were aligned by NCBI BLAST tools and MEGA6.0 software to study the variation of amino acid sites.The eukaryotic expression vectors pEGFP-N1-nef derived from H-TC and N-TC were constructed and transfected into U87 cells to observe green fluorescence.At the same time,the expression of Nef protein,LC3-Ⅱ and p62 protein in U87 cells were detected by Western blot,and the effects of different sources of Nef on autophagy of U87 cells were analyzed.Results The nef genes were amplified by PCR and clone vectors pMD-19T-nef of H-TC and N-TC were successfully constructed.The sequencing confirmed that they were HIV-1B subtypes.The amino acid sequence analysis showed that there were differences between H-TC and N-TC key amino acid sites.The recombinant plasmid pEGFP-N1-nefwas successfully constructed and expressed Nef protein in U87 cells.Western blot analysis showed that the expression of LC3-Ⅱ protein was significantly different among groups (F =11.764,P =0.001).There was no significant difference in the expression of LC3-Ⅱ between cell control and plasmid control(P=0.169).The content of LC3-Ⅱ was low in the two groups of cells,which could not be detected by Western blot.The expression of LC3-Ⅱ in H-TC,N-TC and positive control CQ group increased,compared with blank control or blank vector.The difference was statistically significant (P=0.017,P=0.039,P=0.031),and the expression of LC3-Ⅱ in H-TC group was higher than that in N-TC group (P =0.023);the expression of p62 protein in H-TC,N-TC and positive control CQ group was higher than that in blank control or blank vector group,but there was no significant difference between groups (F=2.049,P =0.163).Conclusions HIV-1 Nef amino acid sites in the central nervous system of patients with HAD and non-HAD were different,and their effects on autophagy of U87 cells were different.The expression of LC3-Ⅱ,an autophagic marker protein,was more strongly induced by Nef from H-TC.

Objective:To analyze the differential expression of silent information regulator transcript-1 (SIRT1) in tissues and cells of nasopharyngeal carcinoma (NPC), to explore the effects of SIRT1 on the proliferation and migration of NPC cells, as well as the effects on and mechanisms of lipid metabolism in NPC cells.Methods:Experimental subjects: In this study, tissue specimens were obtained from patients who visited the Department of Otolaryngology and performed nasopharyngeal tissue biopsy in the Affiliated Hospital of Nantong University from 2019 to 2020. Among them, 6 cases were male, 6 cases were female, age range: 27-72 years old, including 7 cases of NPC diagnosed by pathology and 5 cases of normal nasopharyngeal mucosa. Experimental methods and outcome measures: Western Blot and quantitative real time polymerase chain reaction (qRT-PCR) were used to detect the protein and mRNA levels of SIRT1. CNE2 cell line was selected for subsequent experiments. Cell viability and migratory ability were evaluated by CCK8, wound healing and Transwell assays respectively. Animal xenograft tumor model was used to explore the role of SIRT1 inhibitor Ex527 on tumor growth in nude mice. Oil red and Bodipy were used to stain intracellular lipids. For the mechanical investigation, the interactions between SIRT1 and hypoxia inducible factor-1α (HIF-1α) were analyzed by immunoprecipitation (IP) and chromatin immunoprecipitation (ChIP). Finally, statistical analysis was performed by SPSS 26.0 software, P<0.05 was considered statistically significant. Results:The levels of SIRT1 protein (1.005±0.168) and mRNA (5.829±2.395) in NPC tissues were higher than those in normal nasopharyngeal mucosa (0.181±0.042,1.995±1.605). Differences were statistically significant ( t values were 6.438 and 2.759, both P<0.05). The mRNA and protein levels of CNE1, CNE2, 5-8F and 6-10B cell lines were also higher than those in normal nasopharynx epithelial cell line NP69. Besides, overexpression of SIRT1 correlated with the proliferation and migration of NPC cells. The tumorigenesis ability of nude mice in the Ex527 group was lower than that in the control group. The low SIRT1 expression reduced the protein level of the key enzymes of liposynthesis in NPC cells, improved the expression of lipolysis enzymes, while HIF-1α overexpression promoted lipid synthesis enzymes in NPC cells. SIRT1 inhibited HIF-1α transcription by enhancing deacetylation levels. The binding ability of HIF-1α to SIRT1 promoter regions decreased when NPC cells were hypoxic. Conclusions:SIRT1 promotes the proliferation, migration and lipid metabolism of nasopharyngeal carcinoma cells, which might be expected to provide new theoretical basis for prognosis judgment and gene therapy.

Objective: To investigate the toxic effect of HIV-1 Vpr protein on neurons. Methods: HIV-1 vpr gene was amplified by nested PCR in four parts of peripheral spleen (SPL) and central nervous tissue meninges (MG) of HIV-associated dementia (HAD) patients and non-HAD patients. Eukaryotic expression vector pEGFP-N1-vpr was constructed. The gene sequence and key amino acid sites were analyzed by BLAST and MEGA6. The expression of Vpr protein in N2a cells was detected by Western-blotting. The effects of Vpr proteins from different sources on the activity and cell cycle of N2a cells were studied by flow cytometry. Results: HIV-1 vpr gene was successfully amplified by PCR. Sequence analysis showed that the vpr gene sequence belonged to HIV-1B subtype. There were amino acid mutations at C-terminal 84, 86 and 87 sites of central Vpr protein from HAD and non-HAD patients. Vpr protein could inhibit the activity of nerve cells, leading to G2 phase arrest. Different sources of Vpr had different intensity of action. Compared with other groups, Vpr protein from the meninges of HAD patients showed stronger inhibition of cell activity and G2 phase arrest ability. Conclusions Variations in key amino acid sites of Vpr protein could cause significant changes in its biological functions, and its significance in the pathogenesis of HAD remains to be further studied.

Objective: To study the sequence characteristics and variation of HIV-1 Vpr gene in different parts of an AIDS dementia complex (ADC) patient and provide basis for the study of the neurologic pathogenesis of HIV-1-assciatd dementia. Methods: Genomic DNA was extracted from peripheral samples (lymph nodes, spleen, liver) and central nervous system (meninges, frontal lobe, temporal lobe gray matter, frontal white matter, basal ganglia cortex) of an ADC patient, The Vpr gene was amplified with nested polymerase chain reaction (PCR). PCR products were cloned into the pMD19-T vector. After transformation into DH5α competent E. coli, five positive clones were sequenced. The phylogenetic tree was built and genetic distance was calculated through MEGA6, and the values of ds/dn was calculated through SNAP, then the changes of the amino acid sites were analyzed. Results: HIV-1 Vpr genes isolated from different tissues of the ADC patient had variations. Vpr HIV-1 gene sequences from central nervous system and peripheral tissues were intercrossed together in the phylogenetic tree. Central nervous system and peripheral HXB2 Vpr had no significant differences in genetic distance. The ds/dn of all the HIV-1 Vpr gene sequences were 3.3749. Conclusions The HIV-1 Vpr sequences were different in the ADC patient, and there were different variations in different parts of the peripheral and central regions. Whether these variations are related to the pathogenesis of ADC remains to be further studied.

Objective: BG-derived HIV-1 Tat protein from an HIV-associated dementia (HAD) patient was expressed in E. coli BL21(DE3) and purified in order to research the effects on human umbilical vein endothelial cells (HUVECs) activity. Methods: The recombinant plasmid pGEX-KG-tat with HIV-1 tat stored in our laboratory was amplified by PCR. The PCR product was cloned into pET-32a-tat. The recombinant plasmid pET-32a-tat was transfected into E. coli, and Tat protein was expressed in BL21(DE3), which was induced by IPTG. Then it was purified by Ni-chelating chromatography column and gel filtration preloaded column, and identified by SDS-PAGE and Western blot(WB). The concentration was determined by BCA Kit. Different concentrations of Tat were added into HUVECs to detect their effects on cell activity by cck-8. Results: The Tat with high purity was efficiently expressed in BL21 (DE3) and obtained by using the Ni-chelating chromatography column and gel filtration preloaded column. The concentration was 0.47 mg/ml by using BCA Kit. As the concentration of Tat increased, HUVECs activity decreased. There was no significant difference in cells viability between negative control with 100 ng/ml and 200 ng/ml group (P>0.05). There was significant difference in cells viability between negative control with 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group (P<0.05). But the difference between 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group was not statistically significant (P>0.05). Conclusions The HIV-1 Tat with biological activity was efficiently expressed in BL21 (DE3), and the activity of HUVECs was significantly decreased when the concentration reached 300 ng/ml.

Objective To examine the changing antimicrobial resistance profile of Enterobacter spp.isolates in 53 hospitals across China from 2015 t0 2021.Methods The clinical isolates of Enterobacter spp.were collected from 53 hospitals across China during 2015-2021 and tested for antimicrobial susceptibility using Kirby-Bauer method or automated testing systems according to the CHINET unified protocol.The results were interpreted according to the breakpoints issued by the Clinical & Laboratory Standards Institute(CLSI)in 2021(M100 31st edition)and analyzed with WHONET 5.6 software.Results A total of 37 966 Enterobacter strains were isolated from 2015 to 2021.The proportion of Enterobacter isolates among all clinical isolates showed a fluctuating trend over the 7-year period,overall 2.5％in all clinical isolates amd 5.7％in Enterobacterale strains.The most frequently isolated Enterobacter species was Enterobacter cloacae,accounting for 93.7％(35 571/37 966).The strains were mainly isolated from respiratory specimens(44.4±4.6)％,followed by secretions/pus(16.4±2.3)％and urine(16.0±0.9)％.The strains from respiratory samples decreased slightly,while those from sterile body fluids increased over the 7-year period.The Enterobacter strains were mainly isolated from inpatients(92.9％),and only(7.1±0.8)％of the strains were isolated from outpatients and emergency patients.The patients in surgical wards contributed the highest number of isolates(24.4±2.9)％compared to the inpatients in any other departement.Overall,≤ 7.9％of the E.cloacae strains were resistant to amikacin,tigecycline,polymyxin B,imipenem or meropenem,while ≤5.6％of the Enterobacter asburiae strains were resistant to these antimicrobial agents.E.asburiae showed higher resistance rate to polymyxin B than E.cloacae(19.7％vs 3.9％).Overall,≤8.1％of the Enterobacter gergoviae strains were resistant to tigecycline,amikacin,meropenem,or imipenem,while 10.5％of these strains were resistant to polycolistin B.The overall prevalence of carbapenem-resistant Enterobacter was 10.0％over the 7-year period,but showing an upward trend.The resistance profiles of Enterobacter isolates varied with the department from which they were isolated and whether the patient is an adult or a child.The prevalence of carbapenem-resistant E.cloacae was the highest in the E.cloacae isolates from ICU patients.Conclusions The results of the CHINET Antimicrobial Resistance Surveillance Program indicate that the proportion of Enterobacter strains in all clinical isolates fluctuates slightly over the 7-year period from 2015 to 2021.The Enterobacter strains showed increasing resistance to multiple antimicrobial drugs,especially carbapenems over the 7-year period.