Objective To analyze the endoscopic features of patients with the lower esophageal sphincter (LES) and crural diaphragm (CD) separation and explore its significance in hiatus hernia (HH) diagnosis.Methods From January 2011 to June 2012,the data of patients underwent esophagus high resolution manometry (HRM) examination were retrospectively analyzed and of which the patients with LES-CD separation were selected.Patients who received digestive tracts operation or lack of complete endoscopic date were excluded.A total of 93 cases were enrolled for analysis.According to Chicago Criteria,the cases were divided into three groups.There were 21 cases in type Ⅰ group (LES-CD separation＜1 cm),37 cases in type Ⅱ group (LES-CD separation≥l cm and ＜2 cm),35 cases in type Ⅲ group (LES-CD separation≥2 cm).The differences among groups in endoscopic features were analyzed.HRM were taken as golden standard,the sensitivity and specificity of endoscopy in HH diagnosis were analyzed.Measurement data were expressed as x ± s,one-way ANOVA was performed for multi-groups comparisons,LSD method or Dunnett T3 test for pairwise comparisons and chi-square test or Fisher's exact test for two groups' comparisons.Results There were no significant differences in the distance between esophagus gastric junction (EGJ) and foreteeth of the three groups (P＞0.05).Distances between EGJ and diaphragmatic HH pressure impression in type Ⅲ group,type Ⅱ group and type Ⅰ group were (3.57±0.78) cm,(1.89±0.81) cm and (1.14±0.67) cm,respectively and the differences were significant (LSDt=9.26,11.44,3.57; all P＜0.05).The percentage of continuously-open cardia and showing pressure impression of the gastric pouch of type Ⅲ group was higher than that of type Ⅱ group and the percentage of continuously-open cardia and showing pressure impression of gastric poach of type Ⅱ group was higher than that of type Ⅰ group (80.0％,40.5％ and 4.8％,x2 =11.64,29.76 and 8.59; 91.4％,27.0％ and 4.8％,x2 =30.69,40.73 and 4.32' all P＜0.05).The occurrence rates of the His angle blunting and sac of type Ⅲ group were higher than those of typeⅡ group and type Ⅰ group (74.3％ and 77.1％,24.3％ and 24.3％,4.8％ and 4.8％,x2 =17.97 and 25.41,both P＜0.05),there was no significant difference between type Ⅱ group and type Ⅰ group (P＞0.05).The rates of esophagitis of type Ⅲ group and type Ⅱ group were higher than that of type Ⅰ group (71.4％,59.5％ and 14.3％,x2 =17.14 and 11.15,both P＜0.05),there was no significant difference between type Ⅲ group and type Ⅱ group (P＞0.05).There was one case of Barrett's esophagus in each group.The sensitivity and specificity of endoscopy in HH diagnosis were 91.4％ and 81.0％,respectively.Conclusions The distance between EGJ and hiatal pressure impression,continuously-open cardia,the His angle blunting,hiatal pressure impression of the gastric pouch and hernia sac were important indicators of HH endoscopic features.The sensitivity of endosocopy is higher than the spcificity of endosocopy in HH diagnosis.

Objective:To recognize the characteristics of necrotizing fasciitis patients complicated with sepsis and summarize the experience the treatment.Methods:A retrospective study was conducted. The clinical data of 57 patients with necrotizing fasciitis complicated with sepsis admitted to Guangdong Provincial People's Hospital from July 2009 to December 2019 was analyzed by collecting such factors as gender, age, complications, infection sites, pathogens, surgery information, treatment options and outcome. The patients were divided into debridement group ( n = 14) and control group ( n = 43) according to whether the debridement was completed within 48 hours of admission, and the mortality during hospitalization between the two groups was compared. A telephone follow-up had been done to record the long-term outcome of these patients. Results:Among 57 patients with necrotizing fasciitis complicated with sepsis, there were 43 males and 14 females with the average age of (57.9±12.1) years old. Most of the underlying diseases were diabetes mellitus (70.17%), other diseases included hypertension (8.77%), tumor chemotherapy (7.02%), liver disease (hepatitis, cirrhosis, 7.02%), coronary artery heart disease (3.51%), systemic lupus erythematosus (3.51%), etc. Most of the infection site was lower limbs (71.93%). There were 78 pathogens cultured in 57 patients, in which 52 were non-drug resistant bacteria (66.67%), and 26 were drug resistant bacteria (33.33%). There were 40 Gram positive (G +) bacteria (51.28%), 29 Gram negative (G -) bacteria (37.18%), 8 fungi (10.26%) and 1 mixed bacteria (1.28%). Finally, of 57 patients, 46 patients were cured, and 11 patients died with hospital mortality of 19.30%. Among 57 patients, the hospital mortality in the debridement group was significantly lower than that in the control group [0% (0/14) vs. 25.58% (11/43), P < 0.05]. Among the 46 cured patients, 11 had accepted amputations, accounting for 23.91%. In December 2020, 43 patients who were cured (3 patients were lost to follow-up) were followed up by telephone. Twenty-three patients were completely self-care, 9 patients were partly self-care, 8 patients were completely unable to take care of themselves, and 3 patients died. Conclusions:Necrotizing fasciitis with sepsis mostly occurs in people with weakened immunity, and has a high mortality and disability rate. Early identification and active surgical debridement may be the key to improve the treatment effect.

Objective To investigate the potential protective effects of valproic acid (VPA) on gut barrier function after major burn injury in rats and its mechanism.Methods Forty male Sprague-Dawley (SD) rats were divided into sham + normal saline (NS),sham + VPA,scald + NS,and scald + VPA groups,with 10 rats in each group.Rat with 55％ total body surface area (TBSA) third-degree severe-bums model was reproduced by immersing into 80 ℃ water,and the rats in sham groups were given sham-bums by immersing into 37 ℃ water.The rats after severebums were immediately treated with 0.25 mL of 300 mg/kg VPA or NS by subcutaneous injection.Rats were sacrificed at 2 hours and 6 hours after injury,and abdominal aortic blood and ileal tissue were harvested.The levels of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay (ELISA).The intestinal permeability was evaluated by fluorescein isothiocyanate-dextran (FITC-dextran) determination.The histomorphological changes in gut barrier were evaluated by Chiu grading system.Levels of acetylated lysine at the ninth position of histone 3 protein (Ac-H3K9),hypoxia-inducible factor 1α (HIF-1α),zona occludens 1 (ZO-1) and myosin light chain kinase (MLCK) were determined by immunofluorescence staining and Western Blot.Results Compared with sham + NS group,rats in scald + NS group showed intestinal mucosal damage 2 hours after bum injury,as well as increased mucosal permeability,protein expression levels of HIF-1 α,VEGF,MLCK,and lowered levels of AC-H3K9 and ZO-1.These changes were much more prominent at 6 hours after injury.VPA treatment significantly attenuated the bum-induced intestinal damage.Compared with scald + NS group,the protective effects in scald + VPA group was not evident at 2 hours after injury;however,intestinal damage was much less severe at 6 hours after injury (Chiu score:2.03 ± 0.27 vs.3.12 ± 0.15),intestinal permeability was significantly decreased [FITC-dextran (μg/L):709 ± 76 vs.1138 ± 75],histone acetylation was enhanced [Ac-H3K9 (gray value):1.55 ± 0.12 vs.0.48±0.12],ZO-1 degradation was significantly inhibited (gray value:0.69 ± 0.12 vs.0.43 ± 0.16),the protein expression levels of VEGF and MLCK were significantly down-regulated [VEGF (ng/mg):51.7±3.7 vs.71.2±4.3,MLCK (gray value):1.98±0.20 vs.2.80±0.24],while the HIF-1 α protein expression levels were significantly reduced at both 2 hours and 6 hours after injury (gray value:2.50±0.39 vs.3.88±0.42 at 2 hours,1.83±0.42 vs.4.42±0.41 at 6 hours,all P ＜ 0.05).Conclusions Severe bum injury can induce histone deacetylation,ZO-1 degradation and intestinal barrier dysfunction.VPA can improve the levels of histone acetylation and ZO-1,and protect intestinal epithelial barrier function.These may probably be mediated through inhibiting HIF-1α and its downstream gene VEGF and MLCK.

Objective: To explore the effects of allogeneic bone marrow mesenchymal stem cells (BMSCs) on polarization of peritoneal macrophages isolated from rats with sepsis induced by endotoxin/lipopolysaccharide (LPS). Methods: (1) BMSCs were isolated, cultured and purified from 5 SD rats with whole bone marrow adherent method. The third passage of cells were collected for morphologic observation, detection of expressions of stem cell surface markers CD29, CD44, CD45, and CD90 with flow cytometer, and identification of osteogenic and adipogenic differentiation. (2) Another 45 SD rats were divided into sham injury group (SI, n=5), LPS control group (LC, n=20), and BMSCs-treated group (BT, n=20) according to the random number table. Rats in groups LC and BT were injected with LPS (5 mg/kg) via tail vein to induce sepsis; rats in group SI were injected with the same amount of normal saline to simulate the damage. At post injury hour (PIH) 1, rats in group BT were given 1 mL BMSCs (2×10⁶/mL) via tail vein injection; rats in another two groups were injected with equal volume of phosphate buffer saline. Five rats in group SI at PIH 24 and in groups LC and BT at PIH 6, 12, 24, and 48 were sacrificed to harvest lung tissue for pathological observation with HE staining. In addition, rats in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48 were simultaneously performed with intraperitoneal injection of low-glucose DMEM. Then peritoneal fluid was harvested to culture peritoneal macrophages. Flow cytometer was used to assess the positive expression of cell makers of macrophages including CD68 (making gate), CD11c, and CD206 in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48. Data were processed with one-way analysis of variance and LSD test. Results: (1) The third passage of cells showed uniform fiber-like shape similar to fibroblasts. These cells showed positive expressions of CD29, CD44, CD90 and weak positive expression of CD45. They were able to differentiate into osteoblasts and adipocytes. These cells were identified as BMSCs. (2) At PIH 24, the structure of pulmonary alveoli of rats in group SI was clear and complete with no congestion or inflammatory cell infiltration. At PIH 6, the structure of pulmonary alveoli of rats in groups LC and BT was clear with a small amount of inflammatory cell infiltration, slight congestion and pulmonary interstitial thickening. At PIH 12, the inflammatory responses in lung tissue of rats in group LC were more severe than those in group BT with a large amount of inflammatory cell infiltration, serious congestion, and obvious pulmonary interstitial thickening. The pathological results of rats in group BT at PIH 12 was consistent with the results at PIH 6. At PIH 24, the pathological results of rats in groups LC and BT were similar to the results at PIH 12. At PIH 48, the structure of pulmonary alveoli tissue of rats in group LC was still severely disrupted, with a large number of inflammatory cell infiltration and congestion in lung tissue, but pulmonary interstitial thickening was slightly alleviated than before. The condition of rats in group BT nearly recovered to that in group SI. (3) At PIH 24, the positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(83±10)%] was close to that in group BT [(87±7)%, P>0.05], and they were both significantly higher than the rate in group SI [(55±12)%, with P values below 0.01]. The positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(59±11)%] at PIH 48 was close to that in group SI at PIH 24 (P>0.05), and they were both significantly higher than the rate in group BT [(20±11)%] at PIH 48 (with P values below 0.01). At PIH 24, the positive expression percentages of CD206 in peritoneal macrophages of rats were similar among the three groups (with P values above 0.05). The positive expression percentage of CD206 in peritoneal macrophages of rats in group SI at PIH 24 was close to that in group BT at PIH 48 (P>0.05), and they were both significantly lower than the percentage in group LC at PIH 48 (with P values below 0.01). Conclusions BMSCs can reduce the pathological inflammatory responses in the lung of rats with sepsis and inhibit peritoneal macrophages from polarizing into M1 phenotype, whereas they can not promote macrophages to polarize into M2 phenotype.

OBJECTIVETo investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on secretion of inflammatory cytokines in macrophages stimulated by lipopolysacharide (LPS).

METHODSRat BMSCs and macrophages were isolated, cultured, and identified. The BMSCs and macrophages, cultured alone or in co-culture, were treated with LPS or PBS or without treatment and tested for interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) concentrations in the supernatants at 1, 3, 5, 7, 12, and 24 h after the treatment using enzyme-linked immunosorbent assay.

RESULTSExposure to LPS caused significantly increased IL-10 and TNF-α concentrations in the supernatant of cultured macrophages but not in BMSC culture. Macrophages co-cultured with BMSCs showed significantly lowered IL-10 and TNF-α secretions in response to LPS exposure as compared with the macrophages cultured alone.

CONCLUSIONBMSCs can reduce LPS-induced secretion of inflammatory cytokines by the macrophages to ameliorate inflammatory reactions.

Animals ; Cells, Cultured ; Coculture Techniques ; Enzyme-Linked Immunosorbent Assay ; Inflammation ; Interleukin-10 ; secretion ; Lipopolysaccharides ; Macrophages ; secretion ; Mesenchymal Stromal Cells ; cytology ; Rats ; Tumor Necrosis Factor-alpha ; secretion

To investigate the effect of venous super drainage applying in free flaps. Methods From June, 2017 to December, 2018, 7 cases who had severe soft tissue injuries were treated with free flap. Cause of injury: 1 electric injuries, 2 tumor-related wounds, 1 deep burns, 2 mechanical trauma, and 1 necrotizing fasciitis. All patients were underwent free flap transplantation. There were 5 cases of anterolateral thigh artery perforator flap, 1 case of superficial circumflex iliac artery perforator flap, and 1 case of first dorsal metatarsal artery perforator flap.The application of venous super-drainage technology was carried out according to needs and specific conditions. Two sets of venous passages were prepared in both recipient and donor site to form a double set of venous reflux super-drainage mode. Blood supply, swelling, exudation, secondary exploration and survival rate of the flap were observed after operation, and regularly followed-up. Results All 7 flaps survived. Venous super drainage technique was ap-plied in 7 cases. No arteriovenous crisis occurred after the operation. The flaps had good blood circulation, slight swelling, less exudation, rapid edema regression and no secondary surgical exploration. Followed-up for 2-18 (average 10.5) months, there was no infection recurred. Flaps survived well, and the donor sites healed well without sensory loss.The flexion and extension function of joint was normal. Conclusion The technique of venous super-drainage prepares 2 sets of venous systems for the free flap in the treatment of vascular pedicle in the free flap repair operation, which is conducive to reducing the venous crisis after flap surgery, reducing flap edema, reducing exudation, reducing secondary surgical exploration and improving the survival rate of the flap.