Objective To explore the mechanisms underlying the mediating effect of regulatory B (Breg)cells in the pathogenesis of pemphigus.Methods Peripheral blood specimens were collected from 48 patients with pemphigus at different degrees of severity and 20 healthy controls.Flow cytometry was conducted to analyze the frequency of Breg(CD19 +CD24hiCD38hi)cells in peripheral CD19+ B cells,enzyme linked immunosorbent assay(ELISA)to determine the titer and subtypes of anti-desmoglein(Dsg)1 and 3 antibodies in sera from these subjects.The relationship of anti-Dsg 1 and 3 antibodies with the frequency of Breg cells was assessed by Spearman's rank correlation test.Results Significant differences were observed in the proportion of Breg cells in CD19+ B cells between patients with pemphigus and healthy controls(11.54％ ± 0.97％ vs.8.19％ ± 0.85％,P =0.04),as well as between patients with acute mild pemphigus and those with moderate and severe pemphigus(5.17％ ± 2.14％ vs.11.38％ ± 5.30％ and 11.17％ ± 5.31％,P =0.042,0.046,respectively).The frequency of Breg cells was positively correlated with the absorbance value for anti-Dsg1 antibodies of IgG4 subclass and IgG4/IgG1 ratio(r =0.527,0.565,respectively,both P ＜ 0.01).Conclusions Breg cells are mainly associated with the production of antibodies in and disease severity of pemphigus,while the actual mechanism of action remains unknown.

Objective To study the changes of antigen-specific T helper type 1 (Th1) cells and Th2 cells in patients with pemphigus vulgaris (PV) at different stages,so as to better understand the roles of autoreactive T cells in PV.Methods The DG3 (96-112) peptide was synthesized.Twenty-four patients with PV and 10 health checkup examinees were included in this study.Peripheral blood mononuclear cells (PBMCs) were obtained from the health checkup examinees and all the patients before treatment and seven patients about one month after treatment,and stimulated with the DG3 peptide of 10 mg/L for different durations.Then,enzyme-linked immunospot (ELISPOT) assay was performed to count the number of DG3-specific IFN-γ+ Th1 cells,IL-4+ Th2 cells and memory B cells.One-way analysis of variance (ANOVA) and t test were done to compare the number of cells between different groups,and Pearson correlation coefficient was used to evaluate the correlation among Th cells,memory B cells and anti-desmoglein 3 (Dsg3) antibody titers.Results In these patients,the male to female ratio was 1.67 ∶ 1,and the average age was (56.59 ± 14.66) years.Compared with the health check-up examinees,the patients with PV showed a higher absolute number of DG3-specific IFN-γ+ Th1 cells (420.18 ± 350.29 vs.145.12 ± 86.56,t =3.25,P ＜ 0.05),but a similar absolute number of specific IL-4+ Th2 cells (366.76 ± 192.44 vs.335.88 ± 164.96,P＞ 0.05) per 5 × 105 PBMCs.The percentage of DG3-specific IL-4+ Th2 cells in Th2 cells was 37.03％ ± 23.44％,and the percentage of IFN-γ+ Th1 cells was 23.62％ ± 16.77％ in peripheral blood of patients with PV.The number of DG3-specific IL-4+ Th2 cells per 5 × 105 PBMCs significantly decreased from 452.82 ± 199.29 before treatment to 241.68 ± 160.60 after treatment in seven patients (t =2.48,P ＜ 0.05).There was no significant correlation between specific Th cells,memory B cells and anti-Dsg3 antibody titers (all P ＞ 0.05).Conclusions The peptide DG3 (96-112) has pathogenic epitopes which could be recognized by specific Th cells of patients with PV.Both antigen-specific IFN-γ+ Th1 cells and IL-4+ Th2 cells play certain roles in the pathogenesis of pemphigns vulgaris,and IL-4+ Th2 cells appear to be more important.

Objective To evaluate the short-and long-term outcomes of laparoscopy-assisted gastrectomy (LAG) for gastric cancer.Methods After studying the patients' demographic data,extent of gastrectomy and lymphadenectomy,as well as differentiation and tumor TNM stage,85 patients who underwent LAG were individually matched to 85 patients who underwent open surgery (OG) between October 2004 and March 2008.The operative time,intraoperative blood loss,postoperative recovery,complications,pathological findings,and follow-up data were compared between the two groups.Results The mean operative time was significantly longer in the LAG group than in the OG group (277 ± 62) min vs.(211 ±46) min,t =7.882,P ＜0.05,whereas intraoperative blood loss was significantly lower (161 ±90) ml vs.(267 ± 141) ml,t =-5.854,P ＜0.05.In addition,there was a significant reduction in the time to first flatus and postoperative hospital stay (3.7 ± 1.3) days vs.(4.2 ± 1.1) days and (10 ± 3) days vs.(12 ± 6) days,respectively t =-2.318,-2.325,P ＜ 0.05.There was no significant difference between the LAG group and OG group with regard to the number of harvested lymph nodes and overall postoperative complications.The 5-year disease-free survival rates and overall survival rates were 76％,78％,respectively,in LAG group and 75％,73％,respectively in OG group (all P ＞ 0.05).Conclusions LAG is suitable and minimally invasive for treating gastric cancer.Compared to OG,the LAG will not increase the risk of recurrence and mortality after surgery.

Objective To evaluate the clinical effects of memory alloy embracing fixators in fixation of the rib fractures and investigate the related surgical indications and surgical techniques.Methods Retrospectively review was conducted on the clinical data of 30 patients with rib fractures treated with memory alloy embracing fixators from October 2010 to April 2011 at General Hospital of Guangzhou Military Command.The number of memory alloy embracing fixators used in operation,the number of fixed positions,and operation time were recorded.The pain scores before and after operation were comparatively studied.Operation efficacy and complications were analyzed.Results Of the 30 patients,the total operation time,number of fixators,and number of fixed ribs were (111.9±48.0) minutes,4.3±2.1 and 3.5±1.3,respectively.Meanwhile,the difference between pre-operative and post-operative pain scores was significant (6.93±0.88) points vs (4.04±0.62) points,P＜0.05).The ambulation perlod was (4.6±1.9) days and length of hospital stay was (27.2±10.8) days.Incisional and thoracic wall hematoma was detected in three patients and pulmonary infection in six post-operatively but none presented intractable chest pain,foreign body rejection or wound infection.Conclusion Memory alloy embracing fixators for rib fractures is reliable,easy,and effective in alleviating pain,improving lung function,reducing the frequency of ventilator use and preventing complications like lung infection.

OBJECTIVETo evaluate the safety and feasibility of laparoscopic gastrectomy for gastric stump cancer.

METHODSClinical and follow-up data of 7 patients who underwent laparoscopic gastrectomy for gastric stump cancer in our department from January 2008 to July 2015 were analyzed retrospectively.

RESULTSThere were 5 male and 2 female patients, with a mean age of (62.1±10.7) years. Initial gastrectomy was performed for gastric cancer in 3 patients and peptic ulceration in 4. The initial surgery was B-II( gastrojejunostomy in 6 patients and Roux-en-Y gastrojejunostomy in 1. Duration between primary gastrectomy and occurrence of gastric stump cancer was ranged from 6-30 years for peptic ulceration, and from 11-15 years for gastric cancer. During the operation, adhesiolysis and exploration to locate the tumor were performed. Following total remnant gastrectomy and lymphadenectomy, intracorporeal anastomosis was accomplished by Roux-en-Y reconstruction. The methods of intracorporeal esophagojejunostomy were end-to-side approach using a circular stapler in 1 patient, side-to-side approach using an endoscopic linear staple in 2 patients, and hand-sewn technique in 4 patients. The operation time was (247.1±17.5) minutes and the intraoperative blood loss was (100.0±30.8) ml without transfusion. The number of retrieved lymph node was 19.1±4.8. The first flatus time, diet resumption time, postoperative hospital stay were (3.3±1.5) days, (3.7±0.8) days, (9.4±2.6) days, respectively. One patient experienced gastrointestinal bleeding that was managed conservatively and ultimately cured. Seven patients were followed up till January 2016. After follow-up from 6 to 38 months, 1 patient died of peritoneal metastasis 17 months after surgery, and 1 patient died of Alzheimer's disease 19 months after surgery. The other 5 patients were still alive without metastasis or recurrence.

CONCLUSIONLaparoscopic gastrectomy for gastric stump cancer is feasible and safe.

Aged ; Anastomosis, Roux-en-Y ; Blood Loss, Surgical ; Female ; Gastrectomy ; Gastric Bypass ; Gastric Stump ; pathology ; surgery ; Humans ; Laparoscopy ; Length of Stay ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Operative Time ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Surgical Stapling

OBJECTIVETo evaluate the feasibility and safety of laparoscopic pancreaticoduodenectomy(LPD).

METHODSData of 66 patients from Sir Run Run Shaw Hospital undergoing LPD from September 2012 to September 2014 were reviewed. There were 44 male and 22 female with the mean age of (58.7±10.3) years and mean body mass index of (23.5±3.9)kg/m(2). Forty-five patients presented the symptoms and four of all had the history of abdominal surgery, including 2 cases of laparoscopic pancreatic surgery.

RESULTSOf 66 patients underwent laparoscopic procedure, 1 patient underwent LPD combined with right hepatic resection, 1 patient underwent laparoscopic distal gastrectomy with LPD, and 1 patient underwent LPD after laparoscopic distal pancreatectomy. The mean operative time was (367±49) minutes. The mean blood loss was(193±126)ml. The rate of overall postoperative complications was 36.4%(24/66), with 4.5%(3/66) of B or C pancreatic fistula and 7.6% (5/66) of bleeding. The mean postoperative hospital stay was (18.9±12.1) days. Mean tumor size was (3.8±2.3) cm, and the mean number of lymph nodes harvested was (20.3±10.9). Forty-severn patients were diagnosed as pancreatic adenocarcinoma(n=18), cholangiocarcinoma(n=7), ampullary adenocarcinoma(n=21), and gastric cancer(n=1), respectively.

CONCLUSIONLPD is feasible and safe under the skilled hand.

Adenocarcinoma ; surgery ; Aged ; Anastomosis, Surgical ; Female ; Humans ; Laparoscopy ; Length of Stay ; Male ; Middle Aged ; Operative Time ; Pancreatectomy ; Pancreatic Fistula ; pathology ; Pancreatic Neoplasms ; surgery ; Pancreaticoduodenectomy ; Postoperative Complications

Objective To study the best formulation and technology of quercetin-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate(PLGA-TPGS) nanoparticles(QPTN) with QT as model drug and PLGA-TPGS as polymer materials by orthogonal tests,and to investigate the in vitro stability of QPTN. Methods To ensure the best formula-tion and technology for preparing QPTN,single-factor test was established to determine the influence of the ratio of quercetin to PLGA-TPGS,the concentration of TPGS as emulsifiers,the ultrasonic power and ultrasonic time to the particle size,drug loading (DL) and entrapment efficiency(EE).According to single-factor test,the factor levels of nanoparticles were set to select the best prescription and technology of QPTN by orthogonal test.The stability of QPTN was examined using the effecting factor test,accel-eration test and long-term test. Results The best formulation and technology of QPTN was that the ratio of quercetin to PLGA-TPGS was 3:10 (W:W) with 0.05% TPGS as emulsifier,the mixed solution was sonicated for 6 min at 200 W.The average particle size,DL and EE of QPTN prepared under the conditions described above were (155.4 ± 2.7) nm,(21.6 ± 1.5)% and (93.7±2.9)% (n=6),respectively.In the in vitro stability test,QPTN showed a good stability at high temperature,high humidity and strong light condition. Conclusion The best formulation and preparation technology of QPTN was selected.QPTN got small particle size,high DL and EE,and good in vitro stability.

Objective To evaluate the specific antibody-producing capacity of locally infiltrating B lymphocytes in lesions of patients with pemphigus vulgaris (PV).Methods Totally,35 patients with PV and 22 healthy controls were enrolled into this study.Skin tissues were resected from blisters or erosions of the patients with PV,and from normal skin of healthy controls.Then,mononuclear cells were isolated from these skin tissues.Flow cytometry was performed to determine the percentages of lymphocytes,CD 19+ B lymphocytes,and desmoglein (Dsg)1-and Dsg3-specific CD19+ B lymphocytes.B lymphocytes isolated from the lesional skin of patients with PV were cultured in vitro.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supernatant.Receiver operating characteristic (ROC) curve analysis was conducted to calculate positive rates of anti-Dsg1 and anti-Dsg3 antibodies.Results The percentages of lymphocytes (17.95％ ± 3.85％) and CD19+ B lymphocytes (4.27％ ± 1.13％) were significantly higher in the lesional skin of PV patients than in the normal skin of healthy controls (7.83％ ± 1.29％,0.61％ ± 0.31％ respectively;t =2.49,U =13.00 respectively,both P ＜ 0.05).Among the CD19+ B lymphocytes in the lesional skin of PV patients,the percentage of CD19qgG+ B cells was (38.33 ± 5.56)％,and percentages of Dsg1-and Dsg3-specific CD19+ B lymphocytes were 12.87％ ± 1.267％ and 10.42％ ± 1.243％ respectively.After the in vitro culture for 6 days,the titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supematant were (4.89 ± 1.56) U/ml and (35.45 ± 13.03) U/ml respectively,with their positive rates being 85％ (17/20)and 95％ (19/20) respectively.Conclusion There are Dsg1-and Dsg3-specific B lymphocytes aggregating in the lesional skin of patients with PV,which can produce anti-Dsg1 and anti-Dsg3 antibodies after in vitro culture.

Objective: To evaluate the safety and feasibility of laparoscopic radical antegrade modular pancreatosplenectomy(Lap-RAMPS) for left-sided pancreatic adenocarcinoma. Methods: Clinical data of total 12 patients underwent Lap-RAMPS for left-sided pancreatic adenocarcinoma at Department of Gastrointestinal and Pancreatic Surgery, Zhejiang Provincial People′s Hospital from March 2016 to August 2017 were reviewed retrospectively.There were 7 male patients and 5 female patients, with median age of 60.5 years old(47-68 years old). Abdominal enhanced CT, pancreatic MRI, PET-CT were performed on all patients to evaluate the lesion and exclude metastasis.Follow-up were done with out-patient clinic or telephone consultancy until October 2017. Results: All patients underwent pure Lap-RAMPS.The medium operative time was 250 minutes(180-445 minutes), and the blood loss was 150 ml(50-500 ml). The medium first flatus time and diet resumption time were 3.0 days(1-5 days) and 3.5 days(1-7 days) respectively.The medium postoperative hospital stay was 9 days(4-18 days). Morbidity occurred in 8 patients with gastric empty delay(n=1), bleeding(n=1), fluid collection(n=3). There was no mortality.The medium overall number of retrived lymph nodes was 15.6 and the positive rate was 41.7%. The R0 rate was 100%.The medium follow-up was 10 months.One patient was diagnosed as liver metastasis after 8 months and accepted chemotherapy.One patient died after 14 months for tumor recurrence and metastasis.Others survived without tumor recurrence or metasitasis. Conclusion Lap-RAMPS is safe and feasible with accepted oncological outcomes for selected left side pancreatic adenocarcinoma under skilled hands.

Objective: To summary the experience of 233 cases of laparoscopic pancreaticoduodenectomy (LPD) performed by a single surgical team. Methods: Data of patients undergoing LPD from September 2012 to October 2016 were reviewed. There were 145 males and 88 females with the mean age of(60.3±13.0)years old, ranging from 19 to 92 years old, and the mean body mass index of (22.8±3.5)kg/m^2, ranging from 16.3 to 36.8 kg/m². There were 195 patients with clinical manifestation and 54 patients who had the history of abdominal surgery. Results: LPD were performed on 233 patients by same surgical team consecutively. The mean operative time was(368.0±57.4)minutes. Mean blood loss was(203.8±138.6)ml. The postoperative morbidity rate was 33.5%, with 6.9% of grade B or C pancreatic fistula and 9.9% of bleeding. The reoperation rate was 5.6%. The mortality during 30 days after operation was 0.9%. Mean postoperative hospital stay was (18.1±11.2)days. Mean tumor size was (3.9±2.4)cm, and the mean number of lymph nodes harvested was 21.3±11.9.One hundred and sixty-three patients were diagnosed as malignant tumor, including pancreatic adenocarcinoma(n=84), cholangiocarcinoma(n=17), ampullary adenocarcinoma(n=55), duodenal adenocarcinoma(n=5), gastric cancer(n=1)and duel cancer (n=1) located in distal stomach and duodenum. Conclusion The key point to make laparoscopic pancreaticduodenectomy a routine safe procedure is to operate the procedure under skilled hands in selected patients via suitable surgical approaches.