Objective: To demonstrate that expression of the CCRSDelta32 protein in PBMCs able to down-regulate surface expression of the HIV-1 coreceptor CCR5 and CXCR4.Methods:CCR5Delta32 gene was amplified from human peripheral blood mononuclear cells(PBMCs)genomic DNA by using PCR, and then cloned into lentiviral vector pLenti6/V5-D-TOPO.Recombinant lentiviral particles were produced by packaging using 293T cells.Human PBMCs were transfected with the constructed recombinant lentiviral particles and the expression of CCR5Delta32 was detected by Western blot.The level of CCR5 and CXCR4 expression on transfected PBMCs was detected by FACS analysis.Results: The recombinant lentiviral vector pLenti-CCR5Delta32 was constructed successfully, and the target protein was expressed in PBMCs.FACS analysis showed that CCR5Delta32 protein expressed in PBMCs was able to down-regulate cell surface expression of CCR5 and CXCR4.Conclusion: This study is expected to be used for the gene therapy on AIDS, which deserves further study.

Objective To explore the effect of low molecular weight heparin (LMWH ) in immunoglobulin A nephropathy (IgAN) .Methods Totally 57 patients with IgAN were randomly assigned to two groups :the control group[(treatment with angio‐tensin‐converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) plus statins and oral anticoagulant ];the observa‐tion group(treatment of the same as the control group but instead LMWH for oral anticoagulant ) .All patients were treated for 6 months .24 h urinary protein excretion ,serum creatinine ,prothrombin time(PT ) ,thrombin time(T T ) and activated partial thrombo‐plastin time(APTT) were measured before treatment and at 1 ,3 ,6 months after treatment respectively .The level of urinary trans‐forming growth factor‐β1 (TGF‐β1 ) and laminin(LN) were assayed with the enzyme linked immunosorbent assay (ELISA) .Results Compared with the control group ,24 h urinary protein excretion ,urinary TGF‐β1 and LN levels in the observation group were all decreased at 1 ,3 ,6 months after treatment (P<0 .05) .After 6‐month treatment ,serum Scr level in the observation group(syub‐group) was decreased significantly compared with the control group (subgroup)(P<0 .05) .There were no significant differences in PT ,TT and APTT before and after treatment between the two groups (P>0 .05) .Conclusion The combination therapy with LM‐WH can further decrease proteinuria and ameliorate the renal function of IgAN .

Objective The increasing prevalence of infectious diseases is threatening the biological safety of donated blood in developing countries .This study was to analyze the prevalence of transfusion‐transmissible infectious related measures among first‐time ,voluntary blood donors from 1999 to 2013 in China .Methods From 1999 to 2013 ,all first‐time donors in the Xi′an Blood Service were screened for hepatitis B virus (HBV) ,hepatitis C virus (HCV) ,human immunodeficiency virus (HIV) and Trepone‐miapallidum (TP) and analyzed by trend test analysis .Results The positive rates of HBV ,HCV ,HIV ,and TP in the 415 657 blood donors were 1 .02% ,0 .55% ,0 .05% ,and 0 .46% ,respectively .The prevalence of HBV and HCV presented a decreased trend .Conclusion HBV infection is the primary threat to the blood safety ,while the increasing prevalence of TP and HIV might al‐so be a potential threat .

Objective To investigate the effects of soluble endoglin(sEng)on nitric oxide (NO)production and endothelial nitric oxide synthase(eNOS)phosphorylation in cultured human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells within 3 passages seeded in culture plates of 96 wells,were stimulated by total culture medium(control group)or sEng (1,10 and 100 μg/L)respectively.Cells and medium were collected after cells were cultured for 6,12 and 24 hours respectively.The concentration of the metabolites of NO in each group was measured by nitrate reductase method.The expression of eNOS and eNOS-Ser(p)1177 were detected by Western blot.The expression of eNOS mRNA in each group was detected by real-time fluorescence reverse transcription-polymerase chain reaction.Analysis of variance,LSD method and pearson correlation were used to compare the difference between groups.Results(1)The concentration of the metabolites of NO in 1,10 and 100μg/L sEng groups was(59.25±1.63),(41.08±2.71)and (30.38±1.63)μmol/L respectively after cultured for 6 hours;(54.98±3.34),(35.00±8.60)and (19.82±3.75)μmol/L for 12 hours; and(46.14±4.93),(30.24±2.08)and(12.78±5.01)μmol/L for 24 hours.There was no significant changes in control group with time going by(F=2.30,P=0.14).The concentration of the metabolites of NO was significantly lower in sEng group,and which had negative correlation with culture time(r=-0.98,P＜0.05)and dose(r=-0.88,P＜0.05).(2)The expression of eNOS in 1,10,100 μg/L sEng groups was 0.71 ± 0.00,0.47 ± 0.00 and 0.32±0.00 after cultured for 6 hours; 0.58±0.00,0.42±0.00 and 0.25±0.00 for 12 hours; and 0.49±0.00,0.33±0.00 and 0.18±0.00 for 24 hours.While the expression of eNOS and eNOS-Ser (p)1177/eNOS had no significant changes in control group with time going by(F=3.59 and 0.37,P=0.09 and 0.80).The expression of eNOS protein and eNOS-Ser(p)1177 decreased significantly in sEng groups,which had negative correlation with culture time(r=0.98 and-0.96,P＜0.05)and dose(r=-0.76 and-0.79,P＜0.05).(3)The expression of eNOS mRNA decreased significantly in sEng groups.Which also had negative correlation with culture time(r=-0.51,P＜0.05)and dose(r=-0.82,P＜0.05).Conclusions sEng might inhibit eNOS activity by blocking 1177 Ser phosphorylation to decrease NO production.

Objective To investigate the expression of transforming growth factor-β1 (TGF-β1),endoglin (Eng) and their mRNA in placenta and superficial myometrium of patients with gestational hypertension or preeclampsia and to explore the role of Eng and TGF-β1 in the pathogenesis of gestational hypertension or preeclampsia.Methods One hundred and ten pregnant women were selected in the Second Affiliated Hospital of Tianjin Medical University from April 2009 to April 2010 who underwent cesarean sections and were divided into four groups:gestational hypertension group (n=30),mild preeclampsia group (n=30),severe preeclampsia group (n=30) and control group (normal pregnant women without labor and perinatal complications,n=20).The tissues of placenta and superficial myometrium were collected during cesarean section.Protein levels of Eng and TGF-β1 were detected by Western Blot.Real-time fluorescence reverse transcription polymerase chain reaction was used to detect the Eng and TGF-β1 mRNA expression.One-way ANOVA was used to compare among groups,Student-Newman-Keuls test was used to compare the differences between groups; relation between groups was analyzed by Pearson relation and linear regression.Results In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng mRNA level was 1.00,1.27±0.58,1.54±0.41 and 1.83±0.35,and the TGF-β1 mRNA level was 1.00,1.64 ± 0.33,1.92± 0.38 and 2.23 ± 0.53 in placenta respectively; those figures changed to 1.00,1.32±0.46,1.59±0.37 and 1.93±0.52,and 1.00,1.71 ± 0.45,1.91 ± 0.51 and 2.37 ± 0.46 in superficial myometrium respectively.The Eng and TGF-β1 mRNA levels of control group were lower than those of the other three groups (P＜0.05).The higher the mRNA level,the more severe the disease (P＜ 0.05).In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng protein expression was 0.11±0.07,0.15± 0.05,0.18 ± 0.06 and 0.43 ± 0.04,and the TGF-β1 protein expression was 0.11 ±0.02,0.26 ± 0.05,0.27± 0.03 and 0.88 ± 0.09 in placenta respectively; those figures changed to 0.14±0.06,0.16±0.04,0.20±0.08 and 0.46±0.05,and 0.15±0.03,0.29±0.06,0.31±0.04 and 0.91 ±0.08 in superficial myometrium respectively.The Eng and TGF-β1 protein levels of control group were lower than those of the other three groups (P＜0.05).The higher the protein level,the more severe the disease (P＜0.05).In the gestational hypertension group,mild and severe preeclampsia group,there were positive correlations between Eng and TGF-β1 protein levels in placenta (r=0.57,0.61 and 0.60 respectively,P＜0.05) and superficial myometrium (r=0.59,0.62 and 0.61 respectively,P ＜ 0.05).Conclusions Eng and TGF-β1 might play a role in pathogenesis of gestational hypertension and preeclampsia.

Objective: To investigate the clinical characteristics of salivary gland tumors and their pathological types. Methods: Data from 2 456 patients with salivary gland tumors diagnosed between January 1973 and December 2018 at Sun Yat-sen Memorial Hospital of Sun Yat-sen University were collected, and their gender, age and tumor pathological type, location, and benign and malignant composition ratios were retrospectively analyzed. Results: Over the 46-year study period, 2 456 patients with salivary gland tumors were treated; 41.9% were female, and 58.1% were male. The peak incidence was found among the 40 to 60 years of age group, in which 593 (24.1%) patients had malignant tumors and 1 863 (75.9%) had benign tumors. The ratio of benign and malignant tumors was 3.1∶1. The top two most common benign tumors were pleomorphic adenoma (58.7%) and Warthin tumors (33.6%). The top two most common malignant tumors were mucoepidermoid carcinoma (27.7%) and adenoid cystic carcinoma (26.1%). The most common sites of benign pleomorphic adenomas were the parotid glands, palate, and submandibular glands. Mucinous epidermoid carcinomas in malignant tumors were common in the parotid glands and small salivary glands. The incidence of salivary gland tumors in this group has increased each year, and this group accounted for 53.3% of the total cases over the past 10 years. Conclusion The number of patients with salivary gland tumors is increasing each year. The total incidence of salivary gland tumors is higher in men than in women. Large salivary gland tumors are mainly benign tumors, and small salivary gland tumors are more common. Polymorphic adenomas, Warthin tumors, and mucoepidermoid carcinomas are the most common tumor types; patients 40~60 years old are most likely to have benign salivary glands and have a high incidence of malignant tumors.

Objective : To analyze the value of virtual surgical planning in the surgical treatment of osteoradionecrosis of the mandible and to provide a reference for clinical practice. .Methods : From September 2017 to June 2018, 13 patients with mandibular osteoradionecrosis were evaluated preoperatively using the 3D virtual surgery software CMF Proplan 2.0. The surgical guide was designed and 3D printed. Bone resection, fibula shaping and bone graft localization were completed during the operation. In some cases, implants were implanted at the same time, and denture restoration was completed 3 to 6 months after surgery. Patients’ general information, perioperative data, and efficacy evaluation were analyzed. Results: All patients underwent surgery successfully. The survival rate of the free fibula musculocutaneous flap was 100% (13/13), and one patient had complications (partial necrosis at the edge of the flap). The follow-up period was 7 to 15 months, and the median time was 10 months. All patients achieved a healing effect. The number of cases with an increase in mouth opening ≥ 1 cm, 0.5 cm ≤ mouth opening increase < 1 cm, and mouth opening increase < 0.5 cm were 5, 6, and 2, respectively. An imaging examination showed that 12 patients had good bone healing, and 1 patient did not completely heal 7 months after operation. The denture restoration was 92.3% (12/13), of which 3 cases were implanted and repaired at the same time. The average chewing efficiency was 56.11% ± 7.12% (42.03%-67.83%). Conclusion Virtual surgical planning is an effective method for the surgical treatment of mandibular osteoradionecrosis, which can reduce the risk of surgery and more effectively perform mandibular shape and function repair.

Objective To propose the blood detection strategies for human immunodeficiency virus (HIV) among blood donors, and provide reference for the detection, early diagnosis and transmission blocking of HIV. Methods A total of 117 987 blood samples from blood donors were screened using the third- and fourth-generation ELISA HIV detection reagents. Western blot analysis was used to verify the reactive results of the third-generation reagent alone, or both the third-generation and fourth-generation reagents. HIV nucleic acid test was carried out for those with negative test results of the third- and fourth-generation reagents. For those with positive results of the fourth-generation reagent only, nucleic acid test followed by a confirmatory test by Western blot analysis was carried out. Results 117 987 blood samples from blood donors were tested by different reagents. Among them, 55 were tested positive by both the third- and fourth-generation HIV detection reagents at the same time, accounting for 0.047% and 54 cases were confirmed HIV-positive by Western blot analysis, and 1 case was indeterminate, then turned positive during follow-up testing. 26 cases were positive by the third-generation reagent test alone, among which 24 cases were negative and 2 were indeterminate by Western blot analysis. The band types were p24 and gp160 respectively detected by Western blot analysis, and were confirmed to be HIV negative in follow-up testing. 31 cases were positive by the fourth-generation HIV reagent alone, among which 29 were negative by nucleic acid test, and 2 were positive according to the nucleic acid test.Western blot analysis was used to verify that the two cases were negative. However, after 2~4 weeks, the results turned positive when the blood sample was retested by Western blot analysis during the follow-up of these two cases. All the specimens that were tested negative by both the third- and fourth-generation HIV reagents were validated negative by HIV nucleic acid test. Conclusion A combined strategy with both third- and fourth-generation HIV detection reagents can play a complementary role in blood screening among blood donors. The application of complementary tests, such as nucleic acid test and Western blot analysis, can further improve the safety of blood supply, thus contributing to the early diagnosis, prevention, transmission and treatment of blood donors potentially infected by HIV.
Humans ; HIV Infections/diagnosis* ; HIV Antibodies ; Blood Donors ; HIV-1 ; Blotting, Western ; Nucleic Acids

Tertiary lymphoid structures (TLS) are ectopic lymphoid structures in cancers that are largely associated with favourable prognosis. However, the prognostic value of TLSs in oral squamous cell carcinoma (OSCC) is largely unknown, and the association between tumour infiltrating lymphocytes (TILs) and TLSs has been rarely explored in OSCC. In this study, associated markers of TLS, including peripheral node address (PNAd) in high endothelial venules, CD20 in B cells and CD3 in T cells, were examined in 168 OSCC patients, and survival analysis was performed between TLS-positive and TLS-negative cohorts. We detected the presence of TILs by staining CD8+ cytotoxic T cells and CD57+ NK cells as well. TLSs appeared as highly organized structures in 45 (26.8%) cases. TLS-positive patients had a better 5-year overall survival (OS) rate (88.9% vs. 56.1%, P < 0.001) and relapse-free survival (RFS) rate (88.9% vs. 63.4%, P = 0.002). Moreover, the presence of TLS was an independent prognostic factor for both the 5-year OS rate (hazard ratio [HR] = 3.784; 95% confidence interval [CI], 1.498-9.562) and RFS rate (HR = 3.296; 95% CI, 1.279-8.490) in multivariate analysis. Furthermore, a higher density of CD8+ T cells and CD57+ NK cells was found in TLS-positive sections than in TLS-negative counterparts (P < 0.001), and their combination provided a higher predictive accuracy (AUC = 0.730; 95% CI, 0.654-0.805). In conclusion, our results suggest that TLS is an independent positive prognostic factor for OSCC patients. These findings provide a theoretical basis for the future diagnostic and therapeutic value of TLSs in OSCC treatment.

The heterogeneity of exhausted T cells (Tex) is a critical determinant of immune checkpoint blockade therapy efficacy. However, few studies have explored exhausted T cell subpopulations in human cancers. In the present study, we examined samples from two cohorts of 175 patients with head and neck squamous cell cancer (HNSCC) by multiplex immunohistochemistry (mIHC) to investigate two subsets of Tex, CD8⁺PD1⁺TCF1⁺ progenitor exhausted T cells (TCF1⁺Tex^prog) and CD8⁺PD1⁺TCF1^- terminally exhausted T cells (TCF1^-Tex^term). Moreover, fresh tumor samples from 34 patients with HNSCC were examined by flow cytometry and immunohistochemistry to further investigate their properties and cytotoxic capabilities and their correlation with regulatory T cells (Tregs) in the tumor immune microenvironment (TIME). mIHC and flow cytometry analysis showed that TCF1^-Tex^term represented a greater proportion of CD8⁺PD1⁺Tex than TCF1⁺Tex^prog in most patients. TCF1⁺Tex^prog produced abundant TNFα, while TCF1^-Tex^term expressed higher levels of CD103, TIM-3, CTLA-4, and TIGIT. TCF1^-Tex^term exhibited a polyfunctional TNFα⁺GZMB⁺IFNγ⁺ phenotype; and were associated with better overall survival and recurrence-free survival. The results also indicated that larger proportions of TCF1^-Tex^term were accompanied by an increase in the proportion of Tregs. Therefore, it was concluded that TCF1^-Tex^term was the major CD8⁺PD1⁺Tex subset in the HNSCC TIME and that these cells favor patient survival. A high proportion of TCF1^-Tex^term was associated with greater Treg abundance.
CD8-Positive T-Lymphocytes ; Head and Neck Neoplasms/therapy* ; Humans ; Immunotherapy/methods* ; Prognosis ; Programmed Cell Death 1 Receptor ; Squamous Cell Carcinoma of Head and Neck/therapy* ; Tumor Microenvironment ; Tumor Necrosis Factor-alpha