Objective To determine the concentration of soluble PD-1 (sPD-1) in the sera of patients with recurrent genital herpes (RCH), and to explore the significance of abnormal expression of sPD-1 in RCH. Methods Serum samples were obtained from 88 healthy blood donors, 74 patients with RCH including 34 cases of outbreak-stage RCH and 40 cases of stable-stage RCH. The serum level of sPD-1 was measured by monoclonal antibody labeling and enzyme linked immunosorbent assay (ELISA). Results A significant difference was observed in the serum level of sPD-1 between the patients with RGH and the blood donors (33.06 ± 17.5 μg/L vs. 53.07 ± 26.3μg/L, P < 0.01) and between the patients with outbreak-stage RGH and those with stable-stage RGH (27.47 ± 12.9 μg/L vs. 37.71 ± 19.6 μg/L, P< 0.01). Conclusions There is a low expression of sPD-1 in patients with RGH, which may be one of the mechanisms underlying the escape of HSV- Ⅱ from immunologic surveillance and development of immunological tolerance.

Objective To clone the full-length of human Gill cDNA and construct the recombinant lentiviral expressing vector pLOX-Gfil for eukaryotic expression,providing a basis for further study on the biological functions of Gfil.Methods Total RNA was isolated from K562 cells,and the full-length Gfil cDNA was amplified by RT-PCR and then ligated with pGEM-T vector after retrieve and purification.The ligation product was transformed into competent cells DH5a.The positive recombinant clones were selected and identified by a complementation,restriction endonuclease digestion.The cloning vector and the lentiviral vector pLOX first digested with BarnH I were ligated and transformed.The enzyme and PCR analyses were performed to confirm the recombinant vector,and then DNA sequence analysis.Results A fragment of 1.2 kb was obtained by RT-PCR.The enzyme and PCR analyses revealed that the correct Gfil cDNA was cloned.The sequence of cloned cDNA was identical to the sequence deposited in GenBank (NM005263).Conclusion Gfil was cloned correctly and the recombinant lentiviral vector pLOX-Gfil for eukaryotic expression was constructed successfully.

OBJECTIVE:To establish a method for the contents determination of triamcinolone acetonide acetate and tretinoin in Dimensional ointment. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-0.1%Phosphoric acid solution(87:13,V/V),detection wavelength was 254 nm(0-5 min)and 350 nm(5-20 min),at a flow rate of 1.0 ml/min,column temperature was 25℃,and volume injection was 10 μl. RESULTS:The linear range was 0.5-5 μg/ml for triamcin-olone acetonide acetate(r=0.999 9)and 4.96-49.6 μg/ml for tretinoin(r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 1%;recoveries were 98.37%-100.03%(RSD=0.58%,n=6)and 98.21%-100.38%(RSD=0.78%,n=6). CON-CLUSIONS:The method is simple,accurate and fast,and suitable for the contents determination of triamcinolone acetonide ace-tate and tretinoin in Dimensional ointment.

Objective To investigate the risk factors of gastrointestinal graft versus host disease(GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method 214 patients receiving allo-HSCT from 2003 to 2012 were enrolled in this study,and assessed on the degree of gastrointestinal GVHD.The effects of the primary diseases status,gender,age,conditioning regimen intensity,donor type,the number of cells positive for the CD34 expression,and the use of anti-lymphocyte immunoglobulin (ALG) in the pretreatment scheme on the occurrence of gastrointestinal GVHD was studied.The responses of different degrees of GVHD to immunotherapy were evaluated.Result Univariate and multivariate analyses revealed that the graft type and the conditioning regimen intensity were the risk factors of gastrointestinal GVHD (P＜0.05).Conclusion Donor type and conditioningregimen intensity may be the main risk factors of gastrointestinal GVHD.

Objective In order to evaluate the possible effects of myeloid-derived suppressor cells (MDSCs) on graft versus host disease (aGVHD) development and clinical outcomes,this study systematically detected the dynamic changes of MDSCs accumulation in patients during the first 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Peripheral blood was obtained from 30 patients and 10 healthy volunteers with heparin anticoagulant tubes for 6 mL.For patients,peripheral blood was collected during the first 100 days after allo-HSCT and MDSCs levels were detected by flow cytometry.For measuring the serum concentrations of IL-6,IL-10,IL-1β,TNF-α,Arg-1,HO-1 and iNOS,samples were analyzed using ELISA kits.Results Patients developing aGVHD were infused with significantly less number of MDSCs [(39.94 ± 8.383) 106/kg]than in those not developing aGVHD [(209.0 ± 57.68) 106/kg,P =0.002 6];Patients developing aGVHD Ⅰ-Ⅱ and patients without aGVHD received significantly greater number of MDSCs [(61.96 ± 13.67) 106/kg and (209.0 ± 57.68) 106/kg] than in those developing aGVHD Ⅲ-Ⅳ [(20.37 ±4.304) 106/kg,P =0.013 9].After allo-HSCT,the mean percentage of MDSCs increased markedly in patients developing aGVHD [(7.725 ± 1.460)％] as compared with those not developing aGVHD [(3.423± 1.044)％,P =0.021 3].The high MDSCs group (＞53.712 × 106/kg) showed more favorable clinical outcomes than in the low MDSCs group (≤53.712 × 106/kg).The 2-year overall survival rate as 100％ in high MDSCs group,and 50％ in low MDSCs group (P =0.001 3).The cumulative incidence of 2-year relapse was 6.250％ and 29.252％ in high MDSCs group and low MDSCs group respectively (P =0.112 3).The cumulative incidence of NRM was significantly lower in high MDSCs group (0％) than in low MDSCs group (49.519％,P=0.001 8).MDSCs frequencies significantly increased in patients developing aGVHD after allo-HSCT.After allo-HSCT,the concentrations of IL-6,IL-10,TNF-α,Arg-1,iNOS and HO-1 were significantly elevated in patients developing aGVHD.Conclusion The number of MDSCs when engraftment may be used as a predictor for the development and severity of aGVHD.MDSCs might be considered as a potential new approach to regulate transplant rejection and achieve long-term survival.

This article reports the investigation of the effect of carvedilol (Car) on T-type calcium current (I(Ca,T)) of noninfarcted ventricular myocytes in rabbit models of healed myocardial infarction (HMI). Rabbits with left anterior descending artery ligation were prepared and allowed to recover for 8 weeks, as HMI group. Animals undergoing an identical surgical procedure without coronary ligation were served as the sham-operated group (sham group). Whole cell voltage-clamp techniques were used to measure and compare currents in cells from the different groups. Noting that I(Ca,T) density in HMI cells increased markedly to -2.36 +/- 0.12 pA/pF (at -30 mV) compared with cells of sham, where little I(Ca,T) (-0.35 +/- 0.02 pA/pF) was observed. Meanwhile, further analysis revealed a significant hyperpolarizing shift of steady-state activation curve of I(Ca,T) in HMI cells, where the time constants of deactivation were prolonged and the time of recovery from inactivation was shortened. Finally, the amplitude of I(Ca,T) was increased. Carvedilol (1 micromol x L(-1)) was found to decrease the amplitude of I(Ca,T) to -1.38 +/- 0.07 pA/pF through inhibiting process of I(Ca,T) activation. Furthermore, carvedilol delayed recovery from inactivation of I(Ca,T) and shortened the time constants of deactivation in HMI cells. This study suggested that the application of carvedilol in HMI cells contributes to the dynamic changes in I(Ca,T) and may account for reduction of incidence of arrhythmia after myocardial infarction.

OBJECTIVE: To establish a rapid method for EBV detection with loop-mediated isothermal amplification (LAMP), and to make it as a clue for early diagnosis of nasopharyngeal carcinoma cancer. METHOD: EBV DNA was fast extracted from samples after boiling, while the whole detection will be finished within an hour with specific amplification of EBV gene by LAMP. RESULT: High specificity was shown from EBV detection of 33 clinical samples. Comparing with PCR, LAMP is more simple and convenient to perform under isothermal conditions, and require no special apparatus, thus, it is more economical and practical. CONCLUSION LAMP analysis of EBV may be an efficient and easy way for clinical diagnosis of nasopharyngeal carcinoma.
DNA Primers ; DNA, Viral ; analysis ; Herpesvirus 4, Human ; genetics ; Humans ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; diagnosis ; Nucleic Acid Amplification Techniques ; Sensitivity and Specificity

AIM:This study was performed to investigate the impact of neutrophil extracellular traps(NETs)on scar formation following urethral trauma.METHODS:(1)Clinical samples were derived from patients of Department of Urology,The First Affiliated Hospital of Fujian Medical University,from June 2021 to December 2022.Levels of NETs in the blood and urine were compared between patients with urethral trauma(n=20)and those without urethral trauma(controls,n=20).The relationship between NETs and scar formation was analyzed.(2)Urethral fibroblasts were isolated from urethral scar tissues,and neutrophils were induced to produce NETs in vitro.The urethral fibroblasts were treated with normal saline,0.5 mg/L NETs,or 1.5 mg/L NETs to investigate the effects of NETs on activation and collagen syn-thesis of urethral fibroblasts.Additionally,a rabbit model of urethral trauma was established and the animals were dioided into four groups to explore the therapeutic potential of deoxyribonuclease I(DNase I)in preventing urethral scar forma-tion:control,operation + transforming growth factor-β1(TGF-β1),operation + normal saline,and operation+DNase I.RESULTS:The level of NETs in urine increased after urethral trauma(P<0.05),but the level of NETs in blood did not change(P>0.05).In the animal models,the urethral scar became more severe as the level of NETs in the urine increased(P<0.05).At the cellular level,NETs promoted the viability,migration,and collagen synthesis of urethral fibroblasts(P<0.05)..Additionally,urethral injection of DNase I after trauma reduced the level of NETs and inhibited the formation of urethral scar tissue in the animal models(P<0.05).CONCLUSION:Infiltration of NETs promotes activation of urethral fibroblasts and scar formation after urethral trauma.

Objective To study the risk of bloodstream infection in relation with peripherally inserted central venous catheter (PICC) in hospitalized non-cancer patients.Methods Clinical data of 172 non-cancer patients admitted to our hospital for PICC were collected.The risk of PICC-related bloodstream infection (CRBSI) in domestic hospitalized non-cancer patients was analyzed and systematically assessed.Results Of the 183 PICCs placed in 172 patients included in this study,61.7％ were placed in general wards,38.3％ were placed in ICU,87.8％ were placed in combination with indwelling urinary catheter,29.7％ were placed in combination with mechanical ventilation.The median PICC indwelling time was 35 days.CRBSI occurred in 6 patients with an incidence of 0.6/1000 PICCs/day.The risk of CRBSI was centralized in domestic tumor patients after PICC.The reported CRBSI was significantly different in hospitalized non-cancer patients (0.26-33.10/100 PICC).Conclusion The risk of CRBSI is lower in hospitalized patients after PICC placement than after traditional central venous catheter placement.Further studies are needed to assess its value in ICU.

Objective The risk of current pancreaticojejunostomy is carefully considered from the perspective of the morphology of remnant pancreas,and we aimed to discuss the clinical outcomes of selecting different pancreaticojejunostomy techniques based on pancreatic morphology.Methods This was a prospective cohort study.The histopathology of remnant pancreatic tissues was categorized into four types based on preoperative radiological images and intraoperative palpation:Type Ⅰ:pancreas with hard texture in palpation,pancreatic atrophy,dilated pancreatic duct larger than 5 mm and remnant pancreatic surface ＜3 cm;Type Ⅱ:pancreas with hard texture in palpation,pancreatic atrophy and mild dilatation of pancreatic duct with the diameter of 3-5 mm and remnant pancreatic surface ＜3 cm;Type Ⅲ:pancreas with slightly hard texture,no atrophy,and normal or slightly dilated pancreatic duct with the diameter of 3-5 mm and remnant pancreatic surface ≥3 cm;Type Ⅳ:pancreas with soft texture,normal morphology and pancreatic duct.Results From January 2008 to August 2017,116 consecutive patients underwent pancreaticoduodenectomy in our center.Among them,10 patients with type Ⅰ underwent classic pancreatic ductal mucosa to mucosa anastomosis.19 patients with type Ⅱ underwent classic end to end invaginated pancreaticojejunostomy.45 patients with type Ⅲ underwent classic end to end invaginated pancreaticojejunostomy with overlapping U sutures;42 patients with type Ⅵ underwent total invaginated pancreaticojejunostomy.The post-operative pancreatic fistula occurred in 6 patients (5.2％) with one patient died.Postoperative bleeding occurred in 10 patients (8.6％),and gastroparesis occurred in 22 patients (19.0％).Overall complication rate was 33.6％.Conclusions Classification of pancreatic morphology based on preoperative radiological images and intraoperative palpation and the selection of corresponding pancreaticojejunostomy technique is theoretically rational and has the advantage of potentially reducing the risk of remnant pancreatic tissue.