Objective To assess the mid-term result of laparoscopy in resection of choledoehal cyst,hepaticojejunostomy and discuss the technical improvement.Methods Retrospectively analyzed the data of 31 patients who had undergone laparoscopic choledochal cyst excision from November 2010 to November 2012.Under laparoscopic guidance,intraoperative cholangiogram was performed,Rorx-Y jejunojejunostomy was performed extracorporeally through umbilical incision,then the dilated bile duct were completely excised and a hepaticojejunostomy was carried out intracorporeally.Results Twenty-nine patients successfully underwent laparoscopic choledochal cyst resection,while 2 patients were converted to open surgery because of severe adhesion.The mean operation time was 260 minutes (range from 200 to 460 minutes).A follow-up of 3 to 27 months were achieved in all patients.Three patients developed early complications:one bile leakage,one incision port hemorrhage and one intestinal torsion.The laboratory tests were normal and no dilation of the bile duct was detected in all patients.Conclusions Laparoscopic total cyst excision with Roux-Y hepaticojejunostomy was effective and safe procedure with satisfied results.

OBJECTIVE:To study inhibitory effects of propofol on PC12 cells injury induced by glutamic acid via mitogen-acti-vated protein kinase/extra-cellular regulated kinase (MAPK/ERK) signal pathway. METHODS:PC12 cells were randomized into normal control group,model group(10 mmol/L glutamic acid),propofol low-concentrations,medium-concentrations and high-con-centrations groups(12.5,25,50 μmol/L+10 mmol/L glutamic acid). The optical density of cells,cell apoptosis,the phosphoryla-tion of ERK1/2 and the expression of c-fos,Bax,Bcl-2 were detected after treated with relevant medicine for 48 h. RESULTS:Compared with normal control group,optical density of cells,the phosphorylation of ERK1/2 and Bcl-2 decreased in model group (P<0.01);apoptotic rate,the expression of c-fos and Bax increased (P<0.01). Compared with model group,optical density of cells,the expression of Bcl-2 and the phosphorylation of ERK1/2 increased in propofol group (P<0.01);apoptosis rate,the ex-pression of c-fos and Bax decreased (P<0.05 or P<0.01). There were statistical significant between the different concentrations (P<0.01). CONCLUSIONS:Propofol can inhibit the apoptosis of PC12 cells induced by glutamic acid,which is associated with the up-regulation of ERK1/2 phosphorylation.

OBJECTIVE:To study the effect and its mechanism of carvedilol on leptin-induced activation and proliferation of LX2 human hepatic stellate cells(HSC-LX2). METHODS:HSC-LX2 with logarithmic growth periods were divided into blank con-trol group,leptin-stimulated group and carvedilol low-concentration,medium-concentration,high-concentration groups(5,10,20μmol/L). Except for the blank control group,other groups were added 0.1 g/L leptin and corresponding concentration of carvedilol. After 24 h,MTT method was used to detect the optical density(OD)value of cells and calculate the proliferation rate. Flow cytom-etry was used to detect the cell cycle and apoptosis. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the α-smooth muscle actin (α-SMA),matrix metalloproteinase inhibition factor 1 (TIMP-1),leptin,leptin receptor mRNA expressions. Western blot method was used to detect phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal trans-duction and transcriptional activator 3 (p-STAT3) protein expressions. RESULTS:Compared with blank control group,OD value of cell was increased in leptin-stimulated group;apoptotic rate was decreased;cells of G0/G1 were decreased;α-SMA,TIMP-1, leptin,leptin receptor mRNA expressions and p-JAK2,p-STAT3 protein expressions were increased (P<0.05). Compared with leptin-stimulated group,OD values of cells were decreased in carvedilol concentration groups;apoptotic rate was increased,and the cells were mainly blocked in G0/G1 phase;α-SMA,TIMP-1,leptin,leptin receptor mRNA expressions and p-JAK2,p-STAT3 protein expressions were decreased(P<0.05)and was concentration-depended(P<0.05). CONCLUSIONS:Carvedilol can inhibit the activation and proliferation of leptin-induced HSC-LX2,promote its apoptosis. The mechanism may associate with down-regulat-ing leptin,leptin receptor gene expression and blocking JAK2/STAT3 signal pathway activation by leptin in cells.

BACKGROUND: Pterygium is associated with local chronic inflammatory responses and chronic stimulation from external factors such as. sunlight and wind dust. Presently. there are various theories concerning the onset mechanism of pterygium, but these theories are not generally accepted.OBJECTIVE: To investigate histopathological chractefistics of pterygium and analyze the mulfipotent stem cell effects on the onset of pterygium DESIGN: An open experiment.SETTING: Zhongshan Ophthalmic Center of Sun Yat-sen University.MATERIALS: Experiments were performed at the State Key Laboratory of Ophthalmology, Sun Yat-sen University from September 2006 to January 2007. 218 pterygial paraffin specimens following clinical and pathological diagnosis were obtained from Pathology Lab of Zhongshan Ophthalmic Center, Sun Yat-sea University.METHODS: Pterygial specimens harvested from clinical operations received morphology, immunohistochemistry and immunofluorescence under a confocal microscope.MAIN OUTCOME MEASURES: Morphology of ptorygium and expressions of CD34, vimentin (VIM), smooth muscle acfin (SMA), S- 100 in pterygium.RESULTS: Changes in morphology: Fibroplasia and neovascularization were the main changes in pterygium. Fibroplasia wasdiverse in different regions, and two main phenomena were observed. First, the tissues arranged tightly like the scleral fiber.Secondly, in some loose region, some of spindle-shape, polygonal, asteroid fibroblast-like cells, arranging loosely, could be seen only. No apparent collagen fibers were identified between them. Immunohistochemistry were positive for CD34 in some region where the fibroblast actively proliferated, whereas fibrocytes in mature fibrous tissue were negatively stained.Immunohistochemisu'y was positive for VIM in a large fraction of fibrocytes, vascular endothelial cells, vessel wall and perithelial cells. SMA staining was positive in basophilous small blocks, spindle or irregular cell cluster. Of the 218 cases, 56 cases had smooth muscle. S-100 staining demonstrated that neurofilament protein and adipocytes were positive. Of the 218 cases, 44 cases had adipose tissue, Immunofluorescence showed that proliferative active cells were positive and stained green under a confocal microscope.CONCLUSION: The fibrous tissues in pterygium originate from mesenchymal stem cells, and can differentiate into smooth muscle and adipose tissue.

Objective:To discussed the diagnostic value of magnetic resonance cholangiopancreatography (MRCP), and the use of laparoscopic surgery in management of patients with choledochal cyst with extrahepatic bile duct anomaly.Methods:Of 330 consecutive patients who underwent laparoscopic choledochectomy at Guangzhou Women and Children's Medical Center from January 2010 to September 2018, there were 23 patients with extrahepatic bile duct anomaly. The data of these patients were retrospectively analyzed. There were 4 males and 19 females, with an average age of 3.2 (range 0.3~9.0) years. According to whether the extrahepatic bile duct anomaly was diagnosed by preoperative MRCP, these patients were divided into the preoperative MRCP diagnosis group and the preoperative MRCP undiagnosed group. The impact of MRCP in diagnosing bile duct anomaly to prevent bile duct injury, on operation time, hospital stay and the types of extrahepatic bile duct anomaly on outcomes of laparoscopic treatment were analyzed.Results:All the 23 patients with choledochal cysts complicated by extrahepatic bile duct anomaly were confirmed at surgery. The incidence of extrahepatic bile duct anomaly was 6.97% (23/330). There were 47.8% of type II AHD (11/23); 36.8% of type III AHD (7/23); 4.3% of type IV AHD (1/23); 17.4% (4/23) of the type with communication with accessory bile duct (CABD). The preoperative MRCP diagnosis group consisted of 14 patients, while the preoperative MRCP non-diagnosis group consisted of 9 patients, including 2 patients without MRCP. The diagnostic rate of MRCP in preoperative diagnosis of extrahepatic bile duct anomaly was 66.7%(14/21). The preoperative MRCP undiagnosed group, when compared with the preoperatives MRCP undiagnosed group, had a significantly higher bile duct injury rate [preoperative MRCP diagnosis group 7.1%(1/14), preoperative MRCP non-diagnosis group 55.6%(5/9)], and a significantly longer operation time [preoperative MRCP diagnosis group(232.6±10.0) min, preoperative MRCP undiagnosed group (278.9±22.45)min], (all P<0.05). Laparoscopic surgery was completed in 22 of 23 patients. One patient was converted to open surgery. AHD reconstruction was needed in 11 patients with type II AHD and 1 patient with type IV AHD. Seven patients with type III AHD did not require any surgical intervention for the anomaly. The 4 patients with CABD underwent simple ligation. Postoperative chyloperitoneum developed in 1 patient, who successfully responded to conservative treatment. Postoperative recovery was uneventful in the remaining patients. At a median follow-up of 2 years (range 1 to 5 years), no further complications, including intrahepatic bile duct dilatation and hepatic atrophy were detected. Conclusions:MRCP was effective in preoperative diagnosis of choledochal cysts with extrahepatic bile duct anomaly. It helped to decrease intraoperation injuries to bile duct anomalies. MRCP was also useful in classifying patients with extrahepatic bile duct anomaly to better preoperatively planning of surgical treatment strategies. Laparoscopic surgery could be completed in the majority of these patients with good postoperative results.

Objective Tricuspid regurgitation is often associated in patients with congenital heart disease. Significant morbidity and mortality are related to tricuspid valve replacement. Tricuspid valve plasty is still a preferred choice. This report deals with our surgical experience in using edge-to-edge valve plasty technique to correct severe tricuspid regurgitation in patients with congenital heart disease. Methods From April 2001 to Mar. 2010, severe tricuspid regurgitation was corrected with a flexible band anuloplasty and edge-to-edge valve plasty technique in 14 patients with congenital heart disease. The age ranged from 7 years to 62 years [average (31.2 ± 16.1 ) years]. Congenital cardiac anomalies include: atrioventricular canal in 5 cases, secundum atrial septal defect in 6 cases, secundum atrial septal defect with mitral valve regurgitation in 2 cases and cor triatriatum in 1 case. Results No hospital death or postoperative morbidity occurred. No or trivial tricuspid regurgitation was present in 11 cases and mild tricuspid regurgitation in 3 cases at discharge. The follow-up ranged from 3 month to 97 months [average (51.6 ± 26.8 ) months]. No tricuspid stenosis was found. No or trivial tricuspid regurgitation was present in 5 cases. Mild tricuspid regurgitation was present in 8 cases, and moderate tricuspid regurgitation in 1 case at the latest followup. Conclusion Edge-to-edge valve plasty is an easy, effective and important procedure to correct severe tricuspid regurgitation in patients with congenital heart disease.

Objective To evaluate X-ray stereotactic localized biopsy with resident metal wire in the diagnosis of breast minutelesions. Methods 28 cases of breast minute lesions were analyzed retrospectively,all cases were non-palpable clinically,but were positive by mammography.Needle biopsies were executed under X-ray stereotactic localization,and a thin metal wire was sited in lesion to instruct resection. Results All of 28 cases with minute lesion were successfully localized and completely resected,localization satisfaction rate was 93% percent.Pathological diagnosis showed benign lesion in 19 cases,and malignant lesion in 9 cases.Among the 9 malignant cases,there were 5 cases of invasive duct carcinoma,1 case of duct carcinoma with early invasion,2 cases of duct carcinoma,and 1 case of medullary carcinoma. Conclusion X-ray stereotactic localized biopsy with resident metal wire can provide accurate localization and precise diagnosis,especially in the diagnosis of occult breast cancer.It is an efficient method in the diagnosis of early breast carcinoma.

Abstrac Objective:To investigate the clinical efficacy of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in the treatment of squamous-cell lung cancer patients with EGFR mutations. Methods:We screened out seven patients who were diag-nosed with EGFR mutations and received EGFR-TKIs, such as gifitinib or erlotinib, from 2,317 squamous-cell lung cancer patients treat-ed at the Beijing Chest Hospital from January 2010 to July 2016. Results:After using EGFR-TKIs, the objective response rate was 42.9%, the disease control rate was 100%, and the median progression-free survival was 6.1 months. Conclusion:EGFR-TKIs exert a certain clinical curative effect on patients with EGFR mutations in squamous-cell lung cancer. However, given that only seven cases are pre-sented in this research group, more cases are needed for further research to verify the above conclusion.

Objective To investigate the role of Rho/ROCK signaling pathway in the protective effects of hydrogen gas (H2) on acute lung injury (ALI) in a mouse model of sepsis.Methods Eighty male C57BL/6 mice were randomly divided into four groups (n =20 per group):sham surgery group,H2 control group (sham + H2 inhalation),sepsis model group and H2 treatment group (sepsis + H2 inhalation).The mouse model of sepsis was created by cecal ligation puncture (CLP),and the mice in sham surgery group didn't undergo cecal ligation and puncture.The mice in the H2 inhalation groups received inhalation of 2％ H2 for 1 hour at 1 hour and 6 hours after CLP or sham surgery,respectively.Ten mice in each group were selected and subjected to Evans blue (EB) test to evaluate the pulmonary endothelial permeability at 24 hours after CLP operation.The rest of 10 mice in each group were sacrificed at 24 hours after CLP operation,the bronchoalveolar lavage fluid (BALF) was collected for the measurement of protein concentration,tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1 β) content,and polymorphonuclear neutrophils (PMN)counts.The lung tissues were obtained to determine the content of malonaldehyde (MDA) and the activity of superoxide dismutase (SOD),the wet/dry lung weight ratio (W/D) was calculated,the lung pathological changes in hematoxylin and eosin (HE) stained sections were evaluated under a light microscope,the activity of Rho/ROCK signaling pathway and expression of zonula occluden 1 (ZO-1) were detected by Western Blot,and the distribution and expression of ZO-1 were also examined by immunofluorescence staining.Results There was no statistical difference in the above indexes between the sham surgery group and the H2 control group.Compared with the sham surgery group,the sepsis group demonstrated significant increases in the concentrations of protein,TNF-α,IL-1 β and PMN counts in BALF,the lung EB and MDA content,W/D ratio,the ratio of Rho-GTP/total Rho,the expressions of ROCK1 and ROCK2,the ratio of phosphorylated-myosine phosphatae targeting subunit 1 (p-MYPT1)/MYPT1,and significant decreases in the lung SOD activity and ZO-1 expression.Compared with the sepsis group,the H2 treatment group showed statistically significant decreases in the concentrations of protein,TNF-α,IL-1 β,PMN counts in BALF [protein (g/L):3.12 ± 0.33 vs.6.37±0.56,TNF-o(ng/L):128.45± 17.33 vs.563.83±61.72,IL-1β (ng/L):75.76± 14.35 vs.245.52±30.56,PMN counts (× 105/L):7.46± 1.34 vs.18.55± 5.73],and permeability of lung [EB concentration (μg/g):73.33±6.98vs.144.83± 12.38],the lung MDA content (mmol/g:3.66±0.53 vs.6.04± 1.13),the lung W/D ratio (5.02± 0.34 vs.7.26 ±0.56),the ratio of Rho-GTP/total Rho,the expressions of ROCK1 and ROCK2,the ratio of p-MYPT1/MYPT1 [Rho-GTP/total Rho:(43.12 ± 4.69)％ vs.(68.82± 5.44)％,ROCK1 (gray value):2.42 ± 0.42 vs.6.03 ± 0.64,ROCK2(gray value):2.56± 0.52 vs.4.85 ± 0.53,p-MYPT1/MYPT1:(57.83 ± 8.67)％ vs.(112.50± 13.43)％],and statistically significant increases in the lung SOD activity (kU/g:18.58± 1.68 vs.13.31±2.20) as well as the expression of ZO-1 (gray value:0.61 ± 0.07 vs.0.32 ± 0.06,fluorescence intensity:0.77 ± 0.06 vs.0.54 ± 0.05;all P ＜ 0.05).Moreover,lung HE staining showed that there were obvious lung injuries in the sepsis group which were alleviated in the H2 treatment group.Conclusion H2 could improve endothelial permeability and suppress inflammation and oxidative stress to alleviate ALI in septic mice through inhibition of Rho/ROCK signaling pathway.

AIM: To study the effects of insulin on the proliferation and function of osteoblasts and the relationship between insulin post-receptor change in osteoblasts and osteoblastic cell growth.METHODS: The effects of different levels of insulin on osteoblasts were assessed by MTT colorimetry.Osteocalcin in medium was measured by RIM.IGF-1 mRNA expression levels were determined by RT-PCR.The concentrations of free IGF-1 protein in serum-free medium were measured by ELISA.In addition,the protein level and phosphorylated protein of P~(44/42)MAPK were determined by Western blotting analysis.RESULTS: Insulin enhanced the proliferation of osteoblasts,depending on its dose and exposure time.Insulin at concentration of 10~(-7) mol/L showed the strongest effect,and the action attained the plateau phase beyond 96 h.The best concentration that stimulated synthesis of osteocalcin by insulin was 10~(-7) mol/L.When the insulin concentration beyond 10~(-7) mol/L,the osteocalcin concentration was decreased.Exposure time had no effect on insulin-stimulated synthesis of osteocalcin of osteoblastic cells.When the concentration of insulin reaches 10~(-6) mol/L,the IGF-1 mRNA expression stimulated by insulin was also decreased.The concentrations of free IGF-1 protein in insulin-stimulated groups were all higher than that in control group(P0.05).Insulin acute stimulation rapidly induced the activity of tyrosine phosphorylation of P~(44/42)MAPK.The degree of tyrosine phosphorylation of P~(44/42)MAPK was increased step by step along with the increasing doses of insulin from 0 to 10~(-7) mol/L(P