To retrospectively analyze the clinical data of 48 patients with allergic bronchopulmonary aspergillosis (ABPA) at Shanghai Pulmonary Hospital.There were 23 males and 25 females with a mean age of (36 ± 15) years.Their clinical manifestations included wheeze,cough,sputum production,sputum plugs,fever,hemoptysis,weight loss,chest pain,weakness and night sweats.They had a high peripheral blood eosinophilia,a higher serum total IgE,a higher level of aspergillosis fumigatus-specific IgE and positive immediate skin-prick test to aspergillus fumigatus.Aspergillus species were detected in sputum samples of 26 patients.Chest computed tomography (CT) was performed in 48 patients.There were patchy infiltrations (n =45),transient infiltrations (n =40),central bronchiectasis (n =35) and mucoid impaction (n =18).Obstructive ventilation dysfunction was confirmed by lung function test.

Objective To investigate the expression of endothelial protein C receptor (EPCR) and its roles in plasma and placenta of patients with early onset severe preeclampsia. Methods Sixty cases of severe preeclampsia women who delivered in Xuzhou Maternity and Child Health Care Hospital from March 2014 to February 2016, were recruited, which included 30 cases with early onset severe preeclampsia (early onset group, gestational week <34 weeks ) and 30 patients with late onset severe preeclampsia (late onset group, gestational week ≥34 weeks). Thirty cases of healthy late pregnant women at the same period (gestational week≥34 weeks) were selected as control group. Immunohistochemistry SP method was applied to detect the expression of in EPCR placenta. Reverse transcription (RT)-PCR was used to detect the expression of EPCR mRNA in placenta. ELISA method was used to detect the levels of soluble EPCR (sEPCR)level in plasma of the pregnant women of the three groups. Results The expression of EPCR in placenta mainly distributed in the membrane and cytoplasm of placental syncytiotrophoblasts and vascular endothelial cells, a few in the cell nucleus. The expression of EPCR in early onset group(57%, 17/30)was significantly lower than that in late onset group (93%, 28/30; χ2=25.165,P=0.001). The expression of EPCR in late onset group had no significant difference from that in control group (97%, 29/30;χ2=0.540,P=0.910). The expression of EPCR mRNA in placenta of early onset group(0.40±0.07)was significantly lower than that in late onset group(0.91±0.06;t=-30.044,P=0.001), while there was no statistical difference of the expression of EPCR mRNA between the late onset group and the control group (0.92±0.07;t=-0.631, P=0.538). Plasma sEPCR level in early onset group, late onset group and control group were (231 ± 11), (124±6)and(121±4)μg/L respectively, which is higher in early onset group than that in late onset group (t=48.080,P=0.001). There was no statistical difference of plasma sEPCR level between the late onset group and the control group(t=2.534,P=0.100). Conclusions The pathogenesis of early onset and late onset severe preeclampsia may be different. Decreased expression of EPCR in placenta may be associated with the pathogenesis of early onset severe preeclampsia.

Objective To investigate the effects of alcohol on hepatitis C virus( HCV) replication and type I interferon signaling pathway in human hepatocytes .Methods Primary hepatocytes were treated with different concentrations of alcohol , and then infected with HCV .The infected cells were collected to measure the level of HCV RNA .The alcohol-treated hepatocytes were also collected to detect the expression of HCV Core, IFN-α, IFN-β, IRF-7, suppressor of cytokine signaling SOCS-2 and SOCS-3 at mRNA and protein levels by real-time quantitative PCR and ELISA or Western blot , respectively .Results Alcohol treatment enhanced HCV infection and replication in primary hepatocytes at concentrations higher than 10 mmol/L in a dose-dependent manner (P<0.05).Treatment with 40 mmol/L of alcohol significantly reduced the expression of IFN-α, IFN-βand IRF-7 at mRNA and protein levels , and increased the expression of SOCS-2 and SOCS-3 at mRNA and protein levels .Conclusion Alcohol treatment could damage the host in-nate immunity in human hepatocytes and promote HCV replication by reducing the expression of type Ⅰinter-feron ( IFN-αand IFN-β) and IRF-7 and increasing the expression of negative regulators including SOCS-2 and SOCS-3.These results demonstrated that the impairment of innate immunity in liver of alcohol abusers might contribute to the enhancement of HCV infection and result in poor therapeutic effect of IFN -α.

The aim of this study was to investigate the influence of opiate abuse on the expression of Toll-like receptor 9 (TLR9) in the peripheral blood mononuclear cells (PBMCs) of HIV-1-infected patients and to elucidate possible mechanisms involved in the enhancement of HIV-1 replication by opiate abuse. A total of 200 participants were enrolled in the study by random selection from methadone treatment centers and voluntary HIV counseling and testing centers in the cities of Nanning, Liuzhou, and Qinzhou. These participants included 50 HIV-positive opiate abusers (Opiates HIV(+) group), 50 HIV-negative opiate abusers (Opiates HIV(-) group), 50 HIV-positive subjects who were not opiate abusers (Non-opiates HIV (+) group), and 50 HIV-negative subjects who were not opiate abusers (Control group). PBMCs were isolated from the peripheral blood samples from the subjects and the expression levels of TLR9 mRNA and protein were determined by q-PCR and western blot respectively. There was no significant difference among the four groups in age, gender, nationality, domicile, marital status, educational background or duration of drug abuse (P > 0.05). The median viral loads of the Opiates HIV(+) were significantly higher than those of the Non-Opiates HIV(+) groups (4.450 x 10(3) and 3.977 x 10(3) copies/mL respectively, P < 0.05). The relative expression levels of TLR9 mRNA in the Opiates HIV(+), Non-Opiates HIV(+), Opiates HIV(-) and Control groups were (2.13 +/- 1.59) x 10(-3), (3.66 +/- 2.22) x 10(-3), (1.96 +/- 1.42) x 10(-3) and (7.66 +/- 4.87) x 10(-3), respectively. The expression of TLR9 mRNA was significantly lower in both HIV-1-infected and -uninfected groups of opiate abusers compared with groups of non-abusers (P < 0.05). There was no significant difference in TLR9 mRNA expression levels between the Opiates HIV(+) group and the Opiates HIV(-) group (P > 0.05). However, in the non-opiate groups, the expression levels of TLR9 mRNA in the HIV(+) group were significantly lower than that of the control group (P< 0.05). Western blot results confirmed that the expression of TLR9 protein was lower in the Opiates HIV(+), Non-Opiates HIV(+), and Opiates HIV(-) groups compared to the control group. These results suggest that opiate abuse can decrease the expression of TLR9 in PBMCs, which may result in the enhancement of HIV-1 infection and replication due to a decline in immune response mediated by the TLR9 pathway.
Adolescent ; Adult ; Female ; HIV Infections ; genetics ; metabolism ; virology ; HIV-1 ; physiology ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Opioid-Related Disorders ; genetics ; metabolism ; Toll-Like Receptor 9 ; genetics ; metabolism ; Young Adult

Objective To investigate the expression of TLR 4 and its downstream factor TNF-αin the patients with human immunodeficiency virus and Mycobacterium tuberculosis ( HIV/MTB) co-infection. Methods A total of 119 subjects including 32 patients with HIV infection (HIV group), 30 patients with HIV/MTB co-infection (HIV/MTB group), 28 patients with MTB infection (MTB group) and 29 healthy subjects ( control group ) were recruited continuously from the Fourth People′s Hospital of Nanning City , Guangxi.The expression of TLR4 in peripheral blood mononuclear cells (PBMCs) from the patients was de-termined by flow cytometry .ELISA was performed to detect TNF-αin plasma samples .The HIV-1 viral load was determined by standard method .Results The mean fluorescence intensity ( MFI) for TLR4 expression in PBMCs from HIV, HIV/MTB, MTB and control groups were 21.62±4.67, 18.29±3.87, 16.79±4.45, and 22.85±5.80, respectively, showing significant differences among four groups (F=8.105, P<0.01). The TLR4 levels in MTB and HIV/MTB groups were significantly lower than those in control group ( both P<0.01) and HIV group (P<0.01, P=0.014).The plasma concentrations of TNF-αin HIV, HIV/MTB, MTB and control groups were 15.892 (10.494-21.646) pg/ml, 13.142 (8.014-22.038) pg/ml, 16.284 (11.916-24.005) pg/ml, and 26.657 (16.321-34.541) pg/ml, respectively, that were significantly dif-ferent from each other (F=4.350, P=0.006).The levels of TNF-αin plasma from patients with HIV and HIV/MTB infection were significantly lower than those of healthy subjects (P=0.009 and P=0.001).The viral load in patients from HIV/MTB group (5.113 ±1.018 copies/ml) was significantly higher than that from HIV group (4.416±1.020 copies/ml) (t=3.449, P=0.001).Conclusion MTB infection might promote HIV replication by inhibiting the expression of TLR 4.HIV infection might increase host′s suscepti-bility to MTB infection by reducing the production of TNF-α.Suppressed expression of TLR and TNF-αpro-duction could contribute to the occurrence of HIV /MTB co-infection .

Objective To investigate the impact of methamphetamine (Meth) on the expressions of macrophage inflammatory protein (MIP)-1α ,MIP-1β ,interleukin (IL)-6 among human immunodeficiency virus(HIV)-infected patients .Methods The investigation was performed among 15 Meth-abuse and HIV-infected subjects (Meth + HIV ) ,15 non-Meth-abuse and HIV-infected subjects (non-Meth + HIV ) ,15 Meth-abuse and HIV-uninfected subjects (Meth) ,and 15 healthy subjects (HC) .CD4 + T lymphocyte counts in peripheral blood were detected by flow cytometry .The HIV viral loads in HIV-infected patients were detected by standard detection method .The levels of plasma MIP-1α ,MIP-1β and IL-6 from four groups were determined by enzyme-linked immunosorbent assay (ELISA ) .Intergroup difference was compared using t-test and interactive analysis was conducted using analysis of variance .Results In HIV-infected patients ,CD4 + T lymphocyte counts in Meth + HIV group was significant lower than non-Meth +HIV group (t= 5 .431 , P< 0 .01) ,whereas HIV viral load in Meth + HIV group was significant higher than non-Meth + HIV group (t= 4 .670 , P < 0 .01) .The levels of MIP-1α ,MIP-1β and IL-6 in Meth +HIV group were (40 .60 ± 9 .84) pg/mL , (47 .35 ± 11 .25 ) pg/mL and (37 .94 ± 11 .44 ) pg/mL , respectively ,and those in non-Meth + HIV group were (31 .31 ± 8 .11) pg/mL ,(39 .40 ± 8 .41) pg/mL and (31 .31 ± 8 .11) pg/mL ,respectively .The levels of MIP-1α ,MIP-1β and IL-6 in Meth + HIV group were all significantly higher than those in non-Meth + HIV group(t = 2 .822 , P= 0 .001 ;t = 2 .192 , P=0 .020 ;t= 1 .831 , P = 0 .043 ,respectively ) .The levels of MIP-1α ,MIP-1β and IL-6 in Meth group were (24 .45 ± 5 .90) pg/mL ,(27 .82 ± 7 .25) pg/mL and (27 .18 ± 8 .57) pg/mL ,respectively ,and those in HC group were (28 .42 ± 5 .79) pg/mL ,(31 .76 ± 9 .04) pg/mL and (23 .28 ± 6 .07) pg/mL ,respectively .But there were no significant differences of the levels of MIP-1α ,MIP-1β and IL-6 between Meth group and HC group(t= 1 .860 , P = 0 .158 ; t = 1 .317 , P = 0 .233 ; t = 1 .438 , P = 0 .228 ,respectively) .There was no association between Meth-abuse and the levels of these cytokines (P> 0 .05) ,neither between HIV infection and the levels of cytokines (P> 0 .05) .Conclusion Meth abuse results in elevated expressions of MIP-1αand MIP-1β ,which indicates that Meth abuse may play a regulating role on promoting HIV infection .

Objective To explore the expressions of Toll-like receptor 2 (TLR2 ) and the downstream proteins in patients with human immunodeficiency virus /Mycobacterium tuberculosis (HIV /M TB) co-infection .Methods A total of 119 subjects were randomly enrolled .The subjects were divided into four groups :HIV group (n = 32) ,HIV /M TB group (n = 30) ,M TB group (n = 28) and healthy control group (n= 29) .Peripheral venous blood was collected and the HIV-1 viral load was determined by standard method .The expression levels of TLR2 mRNA in peripheral blood mononuclear cells (PBMC) were determined by real-time quantitative PCR (qPCR) and mean fluorescent intensity (MFI) of TLR2 protein was detected by flow cytometry .The plasma interleukin (IL)-6 and tumor necrosis factor (TNF)-α levels were measured with enzyme-linked immunosorbent assay kits .The data were statistically analyzed by chi-square test ,students t test ,analysis of variance and rank sum test when appropriate .Results The viral load in HIV /M TB group ([5 .113 ± 1 .018] lg copy/mL ) was significantly higher than that in HIV group ([4 .416 ± 1 .020] lg copy/mL ; t = 3 .449 , P< 0 .01) .The TLR2 mRNA expressions in PBMC
among HIV ,HIV/M TB ,M TB and healthy control groups were 1 .397 ± 0 .601 ,1 .463 ± 0 .702 ,1 .429 ± 0 .630 ,and 0 .970 ± 0 .488 ,respectively ,which was significantly different among the 4 groups (F =4 .197 , P= 0 .007) .The MFI of TLR2 protein expressions on PBMC among HIV ,HIV /M TB ,M TB and healthy control groups were 28 .12 ± 4 .55 ,38 .11 ± 11 .77 ,31 .13 ± 12 .10 and 23 .33 ± 5 .14 ,respectively . The TLR2 protein expression levels were significantly different among 4 groups (F= 13 .976 ,P< 0 .01) . The plasma IL-6 and TNF-α concentrations were significantly different among 4 groups (Z = 19 .088 , 15 .475 ,both P< 0 .01) .The IL-6 concentrations in three patient groups were higher than that in healthy control group ,but the TNF-α concentrations were lower than healthy control group .Conclusions The co-infection of HIV-1 and M TB may enhance the activation of TLR2 signaling pathway ,which leads to the increased expression of IL-6 .

To screen the siRNAs (small interference RNA sequences ) which specifically inhibit the gene expression of TLR2 in human monocyte-derived macrophage , and discuss their prospects on the treatment of HIV at the level of molecular immunology.Methods:We obtained the mRNA sequences of human TLR 2 gene from NCBI gene bank ,then designed three siRNAs by siDESIGNTM software.The siRNA targeting human housekeeping gene GAPDH was used as positive control.The fluorescent labeling missense siRNA sequences (NC-FAM ) was used as negative control.We collected fresh peripheral blood from healthy volunteers and isolated mononuclear cells from the blood samples.The human mononuclear macrophages were then purified from mononuclear cells by utilizing adherence method.Cationic liposome reagent Lipofectamine 2000 was used to mediate siRNAs into the human mononuclear macrophages.The levels of TLR2 mRNA expression of siRNA-transfected monocyte-derived macrophage were determined by q-PCR.Expression of TLR2 protein was determined by Western blot.Results: At 72 h after transfection ,we found that the expression of GAPDH mRNA and protein in positive control group decreased significantly.Also found there existed significant differences between each siRNA group (F=41.957,P<0.001).Compared with negative control ,the relative expression of TLR2 mRNA in all siRNAs groups decreased significantly (P<0.05 ) , and the inhibition rates were 46%, 43%, 43% by three miRNAs respectively.Western blot showed that the expression of TLR 2 protein in siRNAs groups decreased significantly compared with that of control (P<0.05 ).Conclusion: The designed siRNAs in this study could inhibit the expression of TLR 2 gene in human monocyte-derived macrophage ,indicating that mediation of TLR-2 expression by siRNA might be a novel strategy for HIV treatment from the per-spective of molecular immunology.

Objective: To explore the feasibility and clinical effect of autologous fat concentrate combined with split-thickness skin grafts in the repair of venous ulcer. Methods: From January 2015 to December 2018, a total of 16 patients diagnosed with venous ulcer were admitted to our department, and all patients received symptomatic and supportive treatment such as local debridement and dressing change. After the granulation tissue grew well, all patients received autologous adipose concentrate combined with spilit-thickness skin graft for repair. Results: All the grafted skin survived well, and the survival rate of grafted skin at 2 weeks after surgery was 86% to 99%. All patients were followed up to 6 months after surgery. The wounds of all patients were healed during the follow-up period. One patient suffered local ulcer recurrence due to repeated friction after healing and the ulcer recovered after dressing changing treatment. All the other patients had no recurrence during the follow-up period. The scar hyperplasia in the skin transplantation area was not obvious and the healing quality was good. No significant surgical complications occurred. Conclusions Autologous lipoconcentrate combined with split-thickness skin grafts is an effective and safe method in the repair of venous ulcer, and the application of autologous lipoconcentrate can effectively improve the survival rate and healing quality of skin grafts.

Objective:To investigate the method, feasibility and clinical effect of scrotal flap in the immediate repair of penoscrotal Paget disease after excision.Methods:From January 2008 to September 2018, a total of 21 patients (age 42-74) diagnosed with penile scrotal Paget disease were admitted and treated in our department. All patients underwent local expanded resection, and intraoperative frozen section examination confirmed a negative margin. All patients used the scrotal flap or scrotal flap combined with other surgical methods to repair the defect.Results:Scrotal flaps were used in ten cases, scrotal flaps combined with other flaps in seven cases, and scrotal flaps combined with skin grafting in four cases. All the scrotal flaps had good blood supply, two cases had small partial skin graft necrosis, and one case had delayed healing of the flap donor area, and all the others had good wound healing. All patients were followed up for six months to 60 months, with an average follow-up time of 30.6 months. Most of the patients (95.2%, 20/21) were satisfied with the appearance of the penis and scrotum, and all the transferred flaps showed no edema. Local recurrence occurred in four patients within two years. Local recurrence and inguinal lymph node metastasis occurred in one patient within three years with a total local recurrence rate of 23.8% (5/21). Due to the small local recurrence lesion, the recurrence lesion was successfully repaired with the adjacent scrotal flap after expanding local excision, and the patients with inguinal lymph node metastasis received ipsilateral inguinal lymph node dissection.Conclusions:The scrotal flap has the characteristics of a large cutting area, flexible design, convenient transfer of flap, enough blood supply, simple, safe, and effective. Using this method can obtain a satisfactory shape of the penis and scrotum after the operation. Those defects that are difficult to use scrotal flap for a complete repair can be repaired with other flaps or skin grafts. A good therapeutic effect can be obtained as well. For the majority of recurrent lesions, scrotal skin flap can be reapplied.