Objective To prepare and identify anti‐CD47 monoclonal antibodies .Methods The gene fragment of CD47 was am‐plified by polymerase chain reaction and cloned into prokaryotic expressing vector pET‐32a(+ ) .Purified reconstructed protein was used to immunize BALB/c mice .The immunized spleen cells were isolated and fused with Sp2/0 cells .After screened ,hybridomas secreting anti‐CD47 monoclonal antibody were acquired .Biological activities of antibodies were investigated by Western blot and flow cytometry .Results The recombinant CD47 extracellular domain protein was successfully expressed in BL21 ,and certificated by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot .Data of flow cytometry detection demonstrated that the antiserum had high affinity to CD47 protein .Conclusion Recombinant CD47 and its monoclonal antibody ,with high affinity , were successfully prepared ,which could provide reliable tools for the future study of CD47 .

Objective To investigate the effects of hydrogen gas on the myocardial injury in rats with endotoxemia.Methods Forty-eight healthy male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 4 groups (n =12 each ):control group (group C ),hydrogen gas control group (group HC ),endotoxemia group (group E) and hydrogen gas + endotoxemia group (group LH).Endotoxemia was induced by intraperitoneal lipopolysaccharide (LPS) 20 mg/kg in groups E and LH,while the equal volume of normal saline was given in groups C and HC.After LPS administration,the rats were exposed to the air containing 2％ hydrogen gas for 6 h in groups HC and LH,and the rats were exposed to the air for 6 h in groups C and E.Blood samples were taken from the abdominal aorta after 6 h inhalation of hydrogen gas to determine the serum level of cTnI.The hearts were then removed to determine the content of TNF-α and IL-6 and expression of phosphorylated nuclear factor κB ( NF-κB) inhibitory protein (p-IκB-α) and p38 mitogen-activated protein kinase (p-p38MAPK) in the myocardial tissues.Results Compared with group C,no significant change was found in the levels of cTnI,TNF-α and IL-6 and expression of p-IκB-α and p-p38MAPK in group HC ( P ＞ 0.05),and the levels of cTnI,TNF-α and IL-6 were significantly increased and the expression of p-IκB-α and p-p38MAPK was up-regulated in groups E and LH ( P ＜ 0.05).Compared with group E,the levels of cTnI,TNF-α and IL-6 were significantly decreased and the expression of p-IκB-α and p-p38MAPK was down-regulated in group LH ( P ＜ 0.05).Conclusion Hydrogen gas can reduce the myocardial injury in rats with endotoxemia,and inhibition of the p38MAPK and NF-κB pro-inflammatory pathway and reduction of the inflammatory response of myocardial tissues are involved in the mechanism.

Objective To evaluate the value of the tests of mean corpuscular volume (MCV ) ,mean corpuscular hemoglobin (MCH) in the screening of thalassemia .Methods 603 cases were performed MCV and MCH tests and gene detection of thalasse-mia .The results of gene detection were used as reference standard .The sensitivity ,specificity ,accuracy ,positive predictive value and negative predictive value of single or combined results of MCV ,MCH tests were calculated .Results The sensitivity and specificity of single MCV test were 91 .2% and 76 .1% ,single MCH test were 91 .2% and 70 .7% .The sensitivity and specificity of parallel combined tests of MCV and MCH were 94 .8% and 61 .4% ,serial combined tests of MCV and MCH were 88 .4% and 75 .6% .Con-clusion The sensitivity of single MCV or MCH test in the screening of thalassemia is high .Parallel combined tests of MCV and MCH can improve the sensitivity ,and will be widely used in the in the screening of thalassemia .

OBJECTIVETo investigate the effect of hydrogen inhalation on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the underlying molecular mechanisms.

METHODSThirty-two male SD rats were randomly divided into 4 groups (n=8), namely the normal saline group (SA), saline with 2% hydrogen gas inhalation group (SH group), ALI group, and ALI with hydrogen inhalation group (LH group). In the two ALI groups, ALI was induced by intraperitoneal injection of 15 mg/kg LPS. Treatments with inhalation of 2% hydrogen gas for 6 h was administered after the injection of LPS or saline. The concentrations of tumor necrosis factor-α (TNF-α) in the lung tissue and serum were examined with ELISA. The expression of p38 MAPK in the lung tissue was detected by Western blotting..

RESULTSHydrogen inhalation decreased the expression of p-p38 MAPK in the lung tissue, and significantly reduced TNF-α content in the lung tissue and serum of rats with ALI.

CONCLUSIONHydrogen inhalation can decrease the expression of TNF-α in the lung tissue and serum, and this effect may be related with reduced p38 MAPK expression and inhibition of p38 MAPK activation.

Acute Lung Injury ; blood ; chemically induced ; metabolism ; Administration, Inhalation ; Animals ; Hydrogen ; administration & dosage ; Lipopolysaccharides ; Lung ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood ; metabolism ; p38 Mitogen-Activated Protein Kinases ; blood ; metabolism

Objective To analyse pathogenesis, types, diagnosis and operation methods of adult intussusception. Methods Clinical data of 150 patients with adult intussusception were reviewed retrospectively . Results Symptom included paroxysmal bellyache (90 0%), abdominal mass (64 7%), nausea and vomit (58 0%), hematochezia (20 7%), constipation (10%) and symptom triad of bellyache , mass and hematochezia (15 3%). Tumor, inflammation, Meckel′s diverticulum and mobile cecum were main cause for intussusception. All 150 patients received operation, 147 patients were cured and 3 patients died. Conclusions Most patients of adult intussusception had pathological basis. The result of surgical therapy is good.

Objective To summarize the experience in prevention and treatment of iatrogenic bile duct injury Methods Clinical data of 112 cases with iatrogenic bile duct injury in ten hospitals of Songhua river drainage area from January 1978 to January 2003 were analyzed retrospectively Results The main cause of iatrogenic bile duct injury was wrong identifying the anatomy of the Calot′s triangle before cholecystectomy accounting for 55 4% (62/112) Diagnosis depended on clinical features, celiac puncture and imaging examination Ultrasonography was among the most sensitive diagnostic means (diagnostic rate=97 5%) Six types of injury were identified according to their locations and type Ⅲ damage was most common in clinical practice (92/112) The curative rate in this group was 95 5% (107/112) Eighty seven cases (77 7%) underwent Roux en Y choledochojejunostomy, with cure rate of 94 3%(82/87) Conclusion Iatrogenic bile duct injury prevention lies in identifing the topography of extrahepatic bile ducts Roux en Y choledochojejunostomy is usually the therapy of choice

Objective:To evaluate the teaching effect of moral education materials implied in scientific research cases in the teaching of "Experimental Traditional Chinese Medicine (TCM)".Methods:The moral education materials implied in scientific research cases of "Experimental TCM" were extracted and skillfully integrated into the teaching of professional knowledge. A questionnaire survey was conducted with questions as "whether it is good to demonstrate the potential humanistic spirit by case teaching, whether this helps improve your interest in science and whether this teaching form affects your study, life and work attitude in the future" "Ten specific items from scientists' moving deeds that touch students and their recognitions" to assess the teaching effect.Results:95.8 percent of students affirmed this teaching form and thought it helped improve their interest in scientific exploration. 87.5 percent of students considered the humanistic spirit would affect their study, life and work in the future. 77.1-89.6 percent of students held a positive attitude to the ten items derived from the scientists' moving stories. Among these items, the percentages of the two items, "the spirit of being able to endure loneliness, work hard to make contributions selflessly without seeking for rewards or reputations" and "having respect forpeople and their scientific research achievements with courage to challenge the authorities of scientific research" are the lowest and highest, respectively.Conclusion:It is good to apply the moral education materials implied in the scientific research cases in the teaching of "Experimental TCM", which basically achieves the teaching goal, but there is still room for improvement.

Experimental Traditional Chinese Medicine is an emerging discipline that plays an important role in cultivating innovative talents of traditional Chinese medicine (TCM). In recent years, with the rapid development of TCM and the new requirements of positioning, and also combined with the college students' cognitive characteristics, we have revised and republished the "Experimental TCM" (Third Edition) textbook, which focuses on introducing knowledge by adopting relevant scientific research cases. This test book was used in the teaching of undergraduates of batch 2013 in the eight-year program in Shanghai University of Traditional Chinese Medicine. After-class questionnaires showed that this teaching mode, guided by scientific research case, is not only helpful for students to develop their quality and ability of adopting modern experimental methods initially in the study and development of TCM, but also able to spread the great achievements of TCM researches. The teaching mode is also conducive to enhancing students' sense of responsibility for the modernization of TCM. Therefore, it is suggested that the course of Experimental TCM should be promoted in the colleges and universities of TCM.

Objective:To explore the role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in non-alcoholic steatohepatitis (NASH).Methods:The liver tissue samples of 24 patients admitted the Second Affiliated Hospital of Fujian Medical University were selected, including 12 NASH samples from liver biopsy and 12 normal liver tissues from the margin of hepatic hemangioma. The expression of NLRP3, apoptosis-associated speck-like protein (ASC), caspase-1, interleukin (IL)-1β and the content of triglyceride (TG) were detected. Wild-type and NLRP3 -/- C57BL/6 mice were fed with normal diet or methionine-choline deficient diet (MCD) for 8 weeks. The wild-type mice were divided into MCC950 NASH, 0.9% sodium chloride (NaCl) NASH, MCC950 and 0.9% NaCl group, 8 mice in each group, and were fed with MCD diet and treated with MCC950, fed with MCD diet and treated with 0.9% NaCl, fed with normal diet and treated with MCC950, and fed with normal diet and treated with 0.9% NaCl respectively for eight weeks. After eight weeks, the pathologic changes of liver tissues were observed with hematoxylin-eosin staining and immunohistochemistry. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), free fatty acid (FFA), IL-1β and TG in serum were determined by enzyme linked immunosorbent assay. The expression levels of NLRP3, ASC, caspase-1 and IL-1β in liver tissues were examined by Western blotting and real-time quantitative polymerase chain reaction. Primary Kupffer cells were isolated and cultured from the livers of wild-type and NLRP3 -/- mice and divided into control group and palmitic acid group. The expression levels of related proteins in the supernatant of cells culture were detected by Western blotting. Independent sample t test and one-way ANOVA were used for statistical analysis. Results:The expression levels of NLRP3, ASC, caspase-1, IL-1β and the content of TG of the liver tissues of the NASH patients were all higher than those of healthy control group (all P<0.05). The formation of steatohepatitis in hepatocyte of MCD-fed mice was more obvious than that of nomal diet-fed mice, with more hepatocyte ballooning and inflammatory cell infiltration. The expression of NLRP3, ASC, caspase-1, IL-1β, caspase-1 activity and the content of TG in liver tissue of NASH mice were all higher than those of normal diet-fed group (all P<0.05); and serum levels of ALT, AST, IL-1β, and the content of FAA were all higher than those of normal diet-fed group (all P<0.05). The serum levels of ALT, AST, IL-1β and IL-18 of NLRP3 -/- NASH mice were all lower than those of wild-type NASH mice (all P<0.05). The serum level of ALT, the expression of ASC, caspase-1 and IL-1β in liver tissues, and the degrees of liver fibrosis of wild-type MCC950 NASH group were all lower than those of 0.9% NaCl NASH group (all P<0.05). The expression of NLRP3, ASC, caspase-1, caspase-1 activity, and secretion of IL-1β and IL-18 in Kupffer cells from wild-type mouse treated with palmitic acid were all higher than those of the negative control group (all P<0.05). However, the changes of the above indicators in Kupffer cells from NLRP3 -/- mouse were not affected by palmitic acid treatment. Conclusion:NLRP3 blockade can significantly alleviate the liver injury and fibrosis in NASH mice and prevent the development of NASH.

Objective:To observe preventive effect of intestinal stent against anastomotic leakage after rectal cancer operation.Methods:A retrospective cohort study was carried out. Clinical data of 107 patients with low rectal cancer undergoing laparoscopic radical resection from January 2015 to August 2019 were retrospectively analyzed. Intestinal stent was placed intraoperatively in 48 cases and was not placed in 59 cases. Postoperative Wexner score for anal function and incidence of anastomotic leakage were compared between patients with and without intstinal stent.Results:There was no significant differences in age, distance between tumor and the anal verge, operative time and postoperative Wexner score for anal function between the two groups (all P>0.05). After a month of follow-up, the incidence of anastomotic leakage was 16.9% (10/59) in the non-stent group, while no anastomotic leakage was found in the stent group ( P=0.002). Conclusion:Placement of intestinal stent can effectively prevent anastomotic leakage after low rectal cancer surgery.