Objective To investigate the value of remnant stomach-jejunal dual pathways reconstruction after laparoscope-assisted radical proximal gastrectomy in the treatment of upper gastric cancer. Methods Twenty-five patients with upper gastric cancer underwent laparoscope-assisted radical proximal gastrectomy and the remnant distal stomach was preserved for side-to-side remnant stomach-jejunal anastomosis and end-to-side jejuno-jejunal anastomosis to reconstruct dual pathways. Results The mean operation time was (240±35) minutes, the mean number of lymph nodes dissected were 22±5, and all the incised margins were negative. No anastomotic leakage, obstruction or stenosis occurred. All patients received postoperative barium meal examination. A large amount of barium directly entered the jejunum, leaving a small amount of barium entered the jejunnum via the route of remnant stomach-duodenum, and was detained in the remnant stomach for 30-60 minutes. No esophageal reflux of barium was observed. All the patients were followed up for 4-18 months, no reflux esophagitis was detected and the short-term life quality was satisfactory. Conclusions Remnant stomach-jejunal dual pathways reconstruction prevents the reflux esophagitis and dumping syndrome, preserves the pathway of duodenum and promotes the life quality of patients.

Objective:To investigate the sensitivity and the specificity of scorpions amplification refractory mutation system ( ARMS) in comparing with that of direct DNA sequencing in the detection of BRAF gene mutations in patients with papillary thyroid microcarcinoma.Methods:Direct sequencing and ARMS were used simultaneously to detect BRAF mutation status in 56 patients with PTMC.Results:BRAF mutations were identified in 46 cases with a mutation rate of 82.9%by ARMS,while in 18 cases with a mutation rate of 32.1%by direct sequencing.Besides,the sensitivity of ARMS was 100%and that of direct sequencing was 39.1%.There were significant differences of both mutation rate and sensitivity between two methods ( P<0.01 ).Conclusion: Compared to direct sequencing,ARMS gains a higher sensitivity in the detection of BRAF mutations in samples with tiny lesions.

BACKGROUND:Schwann cells are important celllines in the process of repairing peripheral nerve injury, and human amniotic homogenate supernatant is shown to secrete a variety of cytokines, which could promote the proliferation of Schwann cells.
OBJECTIVE:To investigate the effect of different concentrations of human amniotic homogenate supernatant on the proliferation of rat Schwann cell96.
METHODS:Schwann cell96 was cultured with high-glucose DMEM containing 20%fetal bovine serum, and the second generation of Schwann cell96 was applied for experiments. The cultured cells were divided into five groups according to different volume fractions of human amniotic homogenate supernatant (0%, 10%, 15%, 20%, 25%) in the medium.
RESULTS AND CONCLUSION:The total protein concentration of human amniotic homogenate supernatant was 675μg/mL, in which the concentration of epidermal growth factor, basic fibroblast growth factor and vascular endothelial growth factor were respectively (470.625±2.546), (4.121±0.026) and (0.172±0.002) ng/L. At 1-7 days, the cellproliferation rate of the 10%and 15%concentration groups was greater than that in 20%and 25%concentration groups (P<0.05);10%and 15%concentrations promoted cellproliferation, while 20%and 25%concentrations inhibited cellproliferation. There were no significant difference in the viability of Schwann cell96 between the control group and the experimental group (P>0.05). Low concentrations (10%, 15%) of human amniotic homogenate supernatant promote the proliferation of Schwann cell96, while high concentrations (20%, 25%) of human amniotic homogenate supernatant inhibit cellproliferation.

OBJECTIVETo investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.

METHODS(1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured. The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope. (2) Two batches of hAMSCs of the third passage were obtained, then the expression of vimentin of cells was observed with immunofluorescence method, and the expression of cell surface marker CD90, CD73, CD105, and CD45 was detected by flow cytometer. (3) hAMSCs-CS of the third passage at culture hour 72 were collected, and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay. (4) Human fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured. Human fibroblasts of the third passage were used in the following experiments. Cells were divided into blank control group and 10%, 30%, 50%, and 70% hAMSCs-CS groups according to the random number table (the same grouping method below), with 48 wells in each group. Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS), while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS. The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12, 24, 48, and 72, respectively, and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments. (5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4), with 4 wells in each group, at post scratch hour (PSH) 0 (immediately after scratch), 12, 24, 48, and 72, the migration distance of cells was observed and measured with inverted phase contrast microscope. (6) Human fibroblasts were grouped and treated as in (5), with 3 battles in each group, and apoptosis rate of cells was detected by flow cytometer. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, LSD test, and t test.

RESULTS(1) On culture day 3, most hAMSCs were in large form, and spindle-shaped with much prominences like fibroblasts or in flat polygonal shape. hAMSCs of the third passage were spindle-shaped. The expression of vimentin of hAMSCs of the third passage was strongly positive, and the expressions of surface markers CD90, CD73, and CD105 of the cells were positive, while the expression of CD45 of the cells was negative. (2) The content of IGF-Ⅰ, VEGF, EGF, and bFGF in hAMSCs-CS were respectively (11.7±1.0), (316±68), (6.1±0.4), and (1.49±0.05) pg/mL. (3) At culture hour 12-72, the proliferation activity of human fibroblasts in each hAMSCs-CS group was significantly higher than that in blank control group (with P values below 0.01), and the proliferation activity of human fibroblasts in 50% hAMSCs-CS group was the highest. (4) The width of scratch in two groups was nearly the same at PSH 0. The migration distance of cells in 50% hAMSCs-CS group was significantly longer than that in blank control group at PSH 12-72 (with P values below 0.01). (5) The apoptosis rate of human fibroblasts in blank control group was (16.2±2.4)%, which was significantly higher than that in 50% hAMSCs-CS group [(7.4±3.6)%, t=6.710, P<0.01].

CONCLUSIONShAMSCs-CS can promote proliferation and migration of human fibroblasts and inhibit the apoptosis of human fibroblasts.

Amnion ; cytology ; Apoptosis ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; metabolism ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mesenchymal Stromal Cells ; chemistry ; Pregnancy ; Vascular Endothelial Growth Factor A ; metabolism

The efficient,stable delivery of siRNA into cells,and the appropriate controls for non-specific off-target effects of siRNA are major limitations to functional studies using siRNA technology.To overcome these drawbacks,we have developed a single lentiviral vector that can concurrently deplete endogenous gene expression while expressing an epitope-tagged siRNA-resistant target gene in the same cell.To demonstrate the functional utility of this system,we performed RNAi-depleted α-actinin-1 (α-ACTN1) expression in human T cells,α-ACTN1 RNAi resulted in inhibited chemotaxis to SDF-1α,but it can be completely rescued by concurrent expression of RNAi-resistant α-ACTN1 (rr-α-ACTN1) in the same cell.The presence of a GFP tag on rr-α-ACTN1 allowed for detection of appropriate subcellular localization of rr-α-ACTN1.This system provides not only an internal control for RNAi off-target effects,but also the potential tool for rapid structure-function analyses and gene therapy.

OBJECTIVETo develop a simple, rapid, accurate, and cost-effective single-0tube multiplex polymerase chain reaction (PCR) assay, which could be used for molecular screening and prenatal diagnosis, for detection of three commonest deletional alpha-thalassemias (-- (SEA), -alpha (3.7) and -alpha (4.2)) in Chinese population.

METHODSFour groups of primers were designed on the basis of gap-PCR, and the PCR reaction condition was optimized systematically with the purpose of amplifying effectively specific DNA fragments that are indicative of the respective genotypes of these three deletional alpha thalassemias. In addition, a pair of primers was designed to amplify LIS1 3' untranslated region (UTR) fragment for use as a separate control for amplification running. A total of 72 blood and prenatal archival DNA samples with various known alpha thalassemia genes or normal alpha globin gene sequence that had been confirmed by Southern blotting analysis or DNA sequencing were collected to test the specificity of this assay by blind analysis. In addition, DNA samples from nine couples at high risk of alpha thalassemia were also analyzed to evaluate the reliability of this technique in prenatal implementation.

RESULTSHomozygote, heterozygote and double heterozygote of the three commonest deletional alpha thalassemias were well detected simultaneously by this established method. For normal allele, a 2.4 kb amplified band as a systematic control and an alpha (2) gene-specific amplicon of 1.8 kb were produced. Besides the two amplified fragments of normal allele, it was found that a 1.3 kb, a 2.0 kb or a 1.6 kb amplified band could be simultaneously shown for representing --(SEA), -alpha (3.7) and -alpha (4.2) alleles, respectively, in the heterozygous states. In a blind test, this technique accurately detected 100% of the DNA samples previously characterized by Southern blotting or DNA sequencing, and it was successfully applied to prenatal diagnosis of alpha thalassemia in nine at-risk families.

CONCLUSIONThe single-tube multiplex PCR protocol presented in this study is easy-to-handle, rapid, reliable and is cost-effective for detecting --(SEA), -alpha (3.7) and -alpha (4.2) chromosomes, and it is suitable for large-scale population screening and for rapid molecular genotyping in clinics.

Asian Continental Ancestry Group ; genetics ; China ; Female ; Heterozygote ; Homozygote ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; Reproducibility of Results ; Sensitivity and Specificity ; alpha-Thalassemia ; diagnosis ; ethnology ; genetics

Objective To assess the effect of radical laparoscopy-assisted gastrectomy (LG) for patients with chronic obstructive pulmonary disease (COPD). Methods Clinical data of 340 gastric cancer patients with COPD undergoing radical gastrectomy with lymphadenectomy at Southwest Hospital, Third Military Medical University between January 2010 and October 2013 were analyzed retrospectively. The clinical outcomes for the 262 patients with COPD who underwent LG (LG group) were compared with those of 78 patients with COPD who underwent open gastrectomy (OG group). During LG, pneumoperitoneum was maintained at an insuffiation pressure of 8 mmHg to 10 mmHg. The primary endpoint was postoperative pulmonary complication (PPC). To predict factors related to PPC, univariate and multivariate logistic analyses were carried out. Results Intraoperative blood loss was significantly less in the LG group [(131.2±14.7) ml] than in the OG group [(246.7±49.0) ml;t=-13.445, P=0.000], but operation time was significantly longer [(220.4±19.1) min vs. (194.2±31.5) min;t=6.877, P=0.000]. The findings showed PPC to be significantly less frequent in the LG group(5.3%,14/262) than in the OG group (15.4%, 12/78)( X2=8.581, P=0.003). The severity of COPD was independent risk factor for PPC(P=0.031, HR=1.456, 95%CI:1.306-1.789). No independent predictor of PPCs was found in type of operative approach (open vs laparoscopic; P=0.126). Conclusion The LG procedure with insuffiation pressure of pneumoperitoneum is tolerated for gastric cancer patients with mild or moderate COPD.

Objective To assess the effect of radical laparoscopy-assisted gastrectomy (LG) for patients with chronic obstructive pulmonary disease (COPD). Methods Clinical data of 340 gastric cancer patients with COPD undergoing radical gastrectomy with lymphadenectomy at Southwest Hospital, Third Military Medical University between January 2010 and October 2013 were analyzed retrospectively. The clinical outcomes for the 262 patients with COPD who underwent LG (LG group) were compared with those of 78 patients with COPD who underwent open gastrectomy (OG group). During LG, pneumoperitoneum was maintained at an insuffiation pressure of 8 mmHg to 10 mmHg. The primary endpoint was postoperative pulmonary complication (PPC). To predict factors related to PPC, univariate and multivariate logistic analyses were carried out. Results Intraoperative blood loss was significantly less in the LG group [(131.2±14.7) ml] than in the OG group [(246.7±49.0) ml;t=-13.445, P=0.000], but operation time was significantly longer [(220.4±19.1) min vs. (194.2±31.5) min;t=6.877, P=0.000]. The findings showed PPC to be significantly less frequent in the LG group(5.3%,14/262) than in the OG group (15.4%, 12/78)( X2=8.581, P=0.003). The severity of COPD was independent risk factor for PPC(P=0.031, HR=1.456, 95%CI:1.306-1.789). No independent predictor of PPCs was found in type of operative approach (open vs laparoscopic; P=0.126). Conclusion The LG procedure with insuffiation pressure of pneumoperitoneum is tolerated for gastric cancer patients with mild or moderate COPD.

A case of laryngeal cancer complicated with Hodgkin's lymphoma treated in the Department of Otolaryngology Head and neck surgery of the First Hospital of Jilin University was reported. Under general anesthesia, right vertical partial laryngectomy, bilateral neck lymph node functional dissection and temporary tracheotomy were performed. No recurrence was found in laryngoscope and color Doppler ultrasound of neck lymph nodes 3 and 5 months after operation.
Humans ; Laryngeal Neoplasms/surgery* ; Hodgkin Disease/complications* ; Neck/pathology* ; Neck Dissection ; Lymph Nodes/pathology* ; Laryngectomy ; Carcinoma/pathology*