Objective To explore the application of laparoscopy in non-traumatic acute abdominal emergency. Methods The authors retrospectively analyzed 201 cases of emergency laparoscopy between March 1999 and May 2002. Results All the 201 patients were unequivocally diagnosed during operation. Laparoscopic operations were successfully completed in 190 cases (139 cases of acute appendicitis, 21 cases of acute cholecystitis, 19 cases of upper gastrointestinal tract perforation, 10 cases of intestinal obstruction and 1 case of common bile duct stones accompanied with acute cholecystitis), with a success rate of 94.5% (190/201). The remaining 11 patients underwent a conversion to open surgery. Conclusions Emergency laparoscopic exploration can not only clarify a diagnosis for acute abdominal emergency with unknown causes but also simultaneously offer a therapeutic method in at least 90% of cases.

BACKGROUND:Articular cartilage is a highly diferentiated tissue, which is very limited in its ability to repair after injury. Stem cel therapy for cartilage repair has completely solved this problem. OBJECTIVE: To investigate the mechanism that diferent growth factors induce the diferentiation of bone marrow mesenchymal stem cels into chondrocytes. METHODS:Passage 5 rat bone marrow mesenchymal stem cels were induced by diferent growth factors and their combinations, including transforming growth factor beta 1 (TGF-β1) group, TGF-β1+insulin growth factor-1 (IGF-1) group, and bone morphogenetic protein 2 (BMP-2)+IGF-1 group, TGF-β1+BMP-2 group, TGF-β1+IGF-1+ BMP-2 group, and blank control group. At 21 days of induction, cels were stained with alcian blue and alizarin red; RT-PCR was employed to detect colagen I mRNA expression. RESULTS AND CONCLUSION:Alcian blue staining showed metachromasia in the cytoplasm and mesenchyma, and proteoglycan was expressed green; alizarin red staining showed no orange calcium nodules. The bone marrow mesenchymal stem cels were preliminarily deduced to diferentiate into chondrocytes, but could not express the cel phenotypes of bone cels. In the blank control group, the expression of colagen I mRNA was negative. Compared with the TGF-β1 group, the mRNA expression of colagen I was lower in the BMP-2+IGF-1 group, but higher in the TGF-β1+BMP-2 group and TGF-β1+IGF-1+BMP-2 group (P < 0.05). Moreover, the expression of colagen I mRNA was highest in the TGF-β1+IGF-1+BMP-2 group (P < 0.05). These findings indicate that TGF-β1 alone is able to induce the chondrogenic diferentiation of bone marrow mesenchymal stem cels, and the combination of TGF-β1, IGF-1 and BMP-2 can play the biggest role to induce the chondrogenic diferentiation of bone marrow mesenchymal stem cels.

Objective To explore the application of plastic pancreatic stents and/or nasal biliary drainage for choledocholithiasis patients having DSBC during Endoscopic Retrograde Cholangiopancreatography (ERCP).Methods Retrospective analysis on clinical data of 57 ERCP cases aiming at choledocholithiasis patients having DSBC from January 2010 to December 2015 has been carried out. According to the guide wire cannulation in an operation, patients are divided into three groups, i.e. plastic pancreatic stents group, nasal biliary drainage group, and plastic pancreatic stents + nasal biliary drainage group, so as to observe the success rate of operation and the occurrence rate of postoperative complications, such as pancreatitis or hyperamylasemia, as well as to compare the differences between the three groups.Results Out of the 57 patients receiving ERCP, 13 patients are in the plastic pancreatic stents group, with two successful operations (15.4%), one case of hyperamylasemia (7.7%), two cases of postoperative pancreatitis (15.4%), one case of fever (7.7%) and one case of hemorrhage (7.7%); 20 patients are in the nasal biliary drainage group, with 20 successful operations (100.0%), no occurrence of hyperamylasemia or postoperative pancreatitis or other complications including hemorrhage and fever; 24 patients are in the plastic pancreatic stents + nasal biliary drainage group, with 19 ERCP operations succeeded at the first attempt (79.2%) and 5 ERCP operations succeeded at the second try (20.8%), as well as 4 cases of hyperamylasemia (16.7%), 2 cases of hemorrhage (8.3%) , no occurrence of fever cases and postoperative pancreatitis. In comparison between the three groups, the occurrence of postoperative pancreatitis and successful rate of operation show a significant difference.Conclusion By adopting plastic pancreatic stents and/or nasal biliary drainage for patients having a dififcult selective biliary cannulation during ERCP, the success rate of operation can be improved, and the occurrence of pancreatitis can also be reduced.

Objective To investigate the effect of miR-141 up-regulation through miR-141 mimic transfection on proliferation,apoptosis and cell cycle of hepatocellular carcinoma (HCC) cells.Methods Real-time polymerase chain reaction (qPCR) was employed to detect differential expression of miR-141 in normal hepatic epithelial cells (L02) and hepatocellular carcinoma cells (HepG2).Furthermore,miR-141 mimics were used to transfect into HepG2 to up-regulate the expression of niR-141 (experimental group).Blank group (CON) and negative control group (miR-NC) were used as control group.qPCR was used to detect expression of miR-141.CCK8 assay was used to detect the proliferation of HepG2 cells.Flow cytometry was used to detect the apoptosis and cell cycle of HepG2 cells.Results The results of qPCR showed that miR-141 was significantly down-regulated in HepG2 cells (0.64 ± 0.13) compared to L02 cells (1.00 ± 0.18),and the difference was significant (P ＜ 0.05).Relative expression of miR-141 was significantly increased after transfection (3.33 ± 0.66),compared to CON group (1.00 ± 0.17,P ＜ 0.05) and miR-NC group (1.08 ±0.14,P ＜0.05).CCK8 assay showed that the absorbance values of HepG2 of miR141 group at 48,72,96 h (0.67 ± 0.07,1.17 ± 0.05,1.36 ± 0.03) were all decreased significantly compared with those in CON group (0.81 ±0.02;1.42±0.03;1.73 ±0.05,all P＜0.05) and miR-NC group (0.78 ±0.01;1.38 ±0.02;1.69 ±0.01,all P ＜0.05).The results showed that miR-141 group [(11.81 ± 0.23)％] had significantly increased apoptosis rate compared to CON group [(4.18 ± 0.18) ％] and miR-NC group [(4.04 ± 0.08) ％],and the difference was statistically significant (P ＜ 0.05).miR-141 group had decreased percentage of S phase cells [(19.89 ± 2.78) ％] compared to CON group [(31.87 ± 1.00) ％,P ＜ 0.05] and miR-NC group [(30.49 ± 1.73) ％,P ＜ 0.05],while miR-141 group had significantly increased percentage of G1 phase cells [(74.74 ±2.03)％] compared to CON group (60.85 ± 1.69) ％ and miR-NC group (60.93 ± 1.95) ％,and the differences were all statistically significant (all P ＜ 0.05).Conclusions miR-141 was in low expression in HepG2 cells.Upregulation of miR-141 could specifically inhibit proliferation and promote apoptosis of HepG2 and change the distribution of cell cycle.

Objective To evaluate laparoscopic hepatectomy for the treatment of primary liver cancer. Methods Nine patients with primary liver cancers at segment Ⅱ, Ⅲ, Ⅴ, Ⅵ and at the edge of the liver underwent laparoscopic partial hepatectomy with hand-assist devices, harmonic scalpel, and Endo-GIA. Results All operations were successful including resection of tumors involving both Ⅱ and Ⅲ segments, and irregular segmentectomy, and 2 cases with additional laparoscopic splenectomy. Surgery lasted for 80～145 min. Intraoperative bleeding was 150～700 ml, with no postoperative complications. Patients were followed-up for 5～25 months with intrahepatic tumor recurrence on 3rd, 4th and 13rd month in one each respectively. Conclusion Hand-assisted laparoscopic partial hepatectomy is a safe and feasible approach for primary liver cancer in clinically selected patients.

OBJECTIVE To understand environmental factors which cause nosocomial infection in ICU. METHODS Air sampling adopted by plain board exposure and hand and object appearance by cotton wool method according to Disinfection Technique Regulation published by Ministry of Health. RESULTS From 289 samples, 182 were qualified and pass ratio was 62.98%. Staphylococcus aureus and mixed opportunistic pathogens were the main bacteria on the air and objects. CONCLUSIONS Through monitoring and analyzing environmental factors of ICU nosocomial infection, to control the prevalence and outbreak of ICU nosocomial infection as well as reduce the chance of infection.

Objective To explore the clinical effects of unilateral placement of plastic stents by ERCP for nonresectable Bismuth type IV hilar cholangiocarcinoma.Methods A prospective study was conducted in 42 patients with nonresectable Bismuth type IV hilar cholangiocarcinoma,who had unilateral insertion of plastic stents in the recent 4 years,All of the patients had successful insertion of single plastic stent by ERCP manipulation.Early results (less than 30 days) of procedure-related complications,mortality,long-term results (greater than 30 days) and survival were observed.Results All of the 42 patients had successful drainage.Mean total bilirubin value decreased from (332.3?163.4)?mol/L to (30.6?18.5)?mol/L,and complete resolution of jaundice was achieved in 61.9% (26/42)of the patients.Early comlications included papilla bleeding due to EST in 4 of 42 patients (9.5%) and acute cholangitis in 10 of 42 patients (23.8%).No procedure-related death occurred.All of the patients needed to have their stents replaced periodically,and the median duration up to the first stent replacement was 65 d,and without significant differences between the two groups in stent insertion to the left and right hepatic duct.Median duration of stent patency was 5.15 months,and median patient survival was 6.5 months,but also no significant differences were found between the two groups.Conclusions Unilateral plastic stent insertion is safe and feasible,can achieve adequate drainage for relatively long time,and can improve life quality and survival for patients with nonresectable hilar cholangiocarcinoma.

Objective To review the basic clinical characteristics and the pathogens and their antimicrobial susceptibilities to neonatal sepsis in very low birth weight infants (VLBWI) and extremely low birth weight infants ( ELBWI) for selection of appropriate antibiotics. Methods A retrospective chart review of 56 cases with neonatal sepsis(early onset neonatal sepsis 3 cases, late onset 53 cases) in VLBWI and ELBWI admitted to the neonatal intensive care unit of Yuying Children's Hospital of Wenzhou Medical College from January 1, 1999 to December 31, 2008 was conducted. The basic clinical characteristics and the results of blood culture and antimicrobial susceptibilities were analyzed. Results Among the 56 cases, the clinical presentations were non-specific. A total of 43 strains of bacteria were isolated, and the most important pathogens responsible for neonatal sepsis in VLBWI and ELBWI were opportunistic pathogenic bacteria. In early onset neonatal sepsis, there was only one culture-proven sepsis that was Chryseobacterium meningosepticum. In the late onset neonatal sepsis cases, the main pathogens of Gram-negative organisms were Klebsiella pneumoniae (33. 3%, 14/42), and the most common Gram-positive organisms were coagulase-negative Staphylococci (26. 2%, 11/42), followed by Enterococcus species (11. 9%,5/42). Furthermore, there were 2 fungal sepsis(4. 8%, 2/42), which were infected by Candida albicans. All of the coagulase-negative Staphylococci were methicillin-resistant coagulase-negative Staphylococci, and they were resistant to common antibiotics and sensitive to vancotnycin and rifampicin. And all of the Klebsiella pneumoniae produced extended-spectrum (Hactamases, which were sensitive only to a few antibiotics such as carbopenems, aminoglycosides and quinolones. Among those 56 cases, 43 patients were cured, 13 died, including six patients who refused any treatments, the mortality rate of neonatal sepsis in VLBWI and ELBWI was 23. 2%. Conclusions The clinical presentations of neonatal sepsis in VLBWI and ELBWI were non-specific, and the most important pathogens were opportunistic pathogenic bacteria, which were multi-drug resistant. Routine blood culture should be taken from infants who are suspected of neonatal sepsis and empirical use of appropriate antibiotics should be initiated as soon as the blood specimen for culture has been drawn. To reduce the occurrence of multi-drug resistant bacteria, we should restrict the use of antibiotics especially the third generation of cephalosporins in neonates as much as possible.

Objective To test whether multiplex ligation-dependent probe amplification(MLPA)could be used for the prenatal detection of the most common aneuploidies of chromosomes 13,18,21,X,and Y.Methods 34 cases including 22 blood samples(12 with trisomy 21,1 with monosomy X,one male witll extra Y and 8 healthy persons),4 cord blood samples with Down syndrome and 8 amniotic fluid samples ( 1 with trisomy 21 and 7 normal fetuses)were recruited into this study.All samples were confirmed by karvotype analysis. DNA was extracted from blood and amniotic lysate was incubated with proteinase K.MLPA was used to determine the relative copy numbers.Results The resuhs were available within 48 h and were concordant with karyotype analysis in all but one case of amniotic fluid that was suggested to be triploid sample 69,XXY by MLPA or contaminated by maternal blood.This sample actually was found containing a number of red blood cells after centfifugation in test. In total,the concordance rate with clinical characteristics was 97.1%.The Ratio values of 13,18,21,X in normal samples were approaching 1.0 except chromosome Y having slightly higher variation in relative copy number.The difference of ratio means between the normal and trisomy 21 samples was statistically significant by one-way ANOVA(F=298.906.P=0.000).Conclusion Computer assisted MLPA with high sensitivity is a rapid,simple,automatic and reliable method for detection of common chromosomal aneuploidies.

Objective To elucidate the interaction between osteoblast of bone marrow microenvironment and leukemia cells,and to investigate the role of osteoblast in the leukemia cells survival and apoptosis and the influence of leukemia cells on the osteoblast.Methods Leukemia cells from AML1-ETO9a-Rac1 mouse leukemia model and osteoblast cells were used.The ratio of GFP+ leukemia cells that co-cultured with or without osteoblast was detected by FACS.In addition,the apoptosis level of leukemia cells was detected by flow cytometry by PI and Annexin Ⅴ labeling.Activation level of PARP was determined by Western-blot.Real-time PCR (RT-PCR) was utilized to detect the mRNA level of TPO,N-cadherin,OPN and Ang1 in osteoblast which was separated from leukemic mice.Results The ratio of GFP+ cells in AE9a-Rac1 leukemia cells co-cultured with osteoblast cell was significantly higher than that of AE9a-Rac1 leukemia cells cultured alone.The apoptotic level of AE9a-Rac 1 leukemia cells cultured alone was significant higher than that of AE9a-Rac 1 leukemia cells in co-culture system.Western blot showed that activated level of PARP in AE9a-Rac1 leukemia cells co-cultured with osteoblast was lower than that cultured alone.RT-PCR result showed that TPO and N-cadherin mRNA levels in primary osteoblast separated from leukemic mice were higher than that from normal mice.Ang1 and OPN mRNA levels of osteoblast from leukemia mice were lower.Conclusion Osteoblast cell can support the survival and inhibit the apoptosis of leukemia cells.Leukemia cells can influence the functions of osteoblast by microenvironment associated cytokines production.