Objective To investigate the improved approach and bile duct approach of laparoscopic biliary tract exploration for the treatment of the elderly patients with common bile duct stones. Methods Ninety-two elderly patients with common bile duct stones were enrolled and randomly divided into the improved group and the conventional group ,with 46 cases in each group. The time of hemorrhage ,the time of common bile duct suture,the recovery time of gastrointestinal function,the time of hospitalization,body temperature,albumin and total bilirubin levels,complications and recurrence of stones were recorded and compared between the two groups. Results The time of hemorrhage,the time of common bile duct suture,the recovery time of gastrointestinal func-tion and the time of hospitalization in the improved group were lower than those in the conventional group(P<0.05, respectively). The body temperature of patients in the improved group was lower than that in the conventional group on 3 days after surgery(P<0.05). The overall incidence of postoperative complications in the improved group was lower than that in the conventional group (P < 0.05). No recurrence of stones was observed in both two groups. Conclusion The improved approach for laparoscopic common bile duct exploration surgery results in less bleed-ing,short common bile duct suture time,rapid postoperative recovery,less body temperature fluctuations and less complications,deserving popularization.

Objective To study the clinical effect of transpedicular vertebral osteotomy spine shortening in treating spinal kyphosis com-plicated with spinal cord nerve dysfunction. Methods A total of 80 patients with spinal kyphosis complicated with spinal cord nerve dys-function in our hospital from May 2013 to June 2014 were enrolled and randomly divided into observation group(n=40) and control group (n=40). The observation group received transpedicular vertebral osteotomy,and the control group received lamina and facet osteotomy. The situation of surgery,vertebral healing and spinal cord function condition,treatment effect between two groups were compared. Results The operation time and postoperative ambulation time of observation group were shorter than those of control group [(76. 52 ± 9. 1) vs (113. 46 ± 13. 44) min,(3. 28 ± 0. 43) vs (5. 67 ± 0. 68) d]. The postoperative bleeding volume,postoperative drainage volume of observation group were less than those of control group [(36. 14 ± 4. 28) vs (55. 23 ± 7. 15) mL,(17. 92 ± 2. 12) vs (29. 64 ± 4. 28) mL]. The Cobb angle and residual urine volume,initial and strong urinary bladder capacity,maximum urinary output of observation group were significantly less than those of control group [(6. 12 ± 0. 68) vs(9. 78 ± 1. 21) mL,(241. 45 ± 28. 56) vs(335. 54 ± 36. 86) mL,(456. 56 ± 51. 78) vs (586. 35 ± 63. 12) mL,(63. 78 ± 7. 24) vs (96. 32 ± 10. 22) mL]. The intervertebral height of observation group was higher than that of control group [(12. 62 ± 2. 81) vs (8. 41 ± 1. 32) mm]. The excellent rate of observation group was significantly higher than that of control group(97. 50%vs 82. 50%). Conclusion Transpedicular vertebral osteotomy spine shortening is helpful to reduce operation wound, pro-mote postoperative recovery,correct kyphotic deformity and improve neurological functionin,improve therapeutic effect.

Objective To establish human choriocarcinoma JeG-3 cell line resistant to floxuridine (FUDR)and describe the characteristics of this FUDR-resistant subline.The thymidylate synthase (TS) expression level in FUDR-resistant subline was also discussed.Methods The FUDR-resistant sub-line JeG-3/FUDRA was established by intermitted exposure to grads increased FUDR.Reversed microscope was used to observe the morphological changes in FUDR-resistant sub-line.Population doubling time was calculated and compared based on the growth curve of these two cell lines,cell cycles and chromosomal ploidy were assayed with flow cytometry methods.The chemo-luminescence assay was used to detect the hormone secretion by two kinds of cell lines.The resistant index (RI) was measured by cell counting kit-8 (CCK-8)assay.Quantitative RT-PCR was used to detect the mRNA expression level of TS and we also detected the TS mRNA expression level in different doses exposed subline.Results The RI of JeG-3/FUDRA was 31.62.Compared with the JeG-3 cell,the FUDR-resistant cell line had gross changes in morphological,cell growth,cell cycles and chromosomal numbers.The ability of human chorionic gonadotrop(hCG) and progesterone secretion was lower in JeG-3/FUDRA subline.The trend of TS mRNA expression was:while exposed to low concentration of FUDR,the TS mRNA expression level was downregulated,then followed the increasing dose of the drug,the expression level of TS mRNA ascended gradually.When the terminal concentration was reached,the expression level of TS mRNA in JeG-3/FUDRA subline was higher than that of JeG-3 cell line (P＜0.05).Conclusions We established the FUDR-resistant subline of JeG-3 successfully.The TS mRNA expression level is stage-related to the different concentration and different phase in FUDR exposure.Our data suggested that TS mRNA expression level may not be used as a biomarker to predict the chemosensitivity in FUDR-based chemotherapy.

Objective To explore the effect of paraspinal approach and posterior median approach for one -stage decompression recon-struction in the treatment of thoracolumbar spine fracture and dislocation .Methods From January 2012 to January 2014, 60 patients with thoracolumbar spine fracture and dislocation who were admitted and treated in our hospital were selected as the research objects .All patients received one-stage decompression reconstruction for treatment .According to the methods of approach , the patients were divided into the ob-servation group and the control group .The 30 cases in observation group were treated by paraspinal approach while the other 30 cases in con-trol group were treated by posterior median approach .Visual analogue score ( VAS) was applied .The status of surgery , recovery of centrum height, changes of Cobb angle as well as the occurrence rate of postoperative complications in the two groups were recorded .Results In the observation group, the operative time, time in bed and length of stay were (89.16 ±11.63) min, (39.75 ±8.69) h and (3.96 ±1.04) d respectively, which were shorter than those in the control group .The intraoperative blood loss was (89.64 ±13.62) mL which was lower than that in the control group and the difference was significant (P<0.05).One week after operation, the anterior and posterior height of centrum in the observation group increased significantly while Cobb angle significantly reduced .Compared with those before the treatment , the difference was significant (P<0.05).The maximum coronary diameter and maximum sagittal diameter of paraspinal muscles in the ob -servation group after the treatment were (48.96 ±5.34)mm and (18.16 ±6.74)mm respectively, which were significantly higher than those in the control group and the difference was significant (P<0.05).The incidence of lumbar and back pain in the observation group was 3.33%which was lower than 23.33%in the control group and the difference was statistically significant (P<0.05).Conclusion To carry out decompression reconstruction through paraspinal approach can reduce the the pain degree of patients and the incidence of lumbar and back pain after operation .

Objective To investigate the state of Th1/Th2 cytokine imbalance in patients with lupus nephritis (LN) and its role in the pathogenesis. Methods Plasma level of interleukin-18 and interleukin-13 in 18 patients with active LN and 16 normal controls were measured by enzyme-linked immunoadsordent assay (ELISA). IL-18 and IL-13 expression in the renal tissues from 18 patients and 6 normal renal tissues were also detected by immunohistochemical assay. The ratio of plasma and renal IL-18/IL-13 was then calculated. Results Plasma levels and renal expression of IL-13 and IL-18 in patients with LN were increased significantly compared to those of normal controls (P0.05). The ratio of renal IL-18/IL-13 was not significantly different among all types of LN and normal controls. The ratio of plasma IL-18/IL-13 was positively correlated with LN renal tissue activity index (AI), but no correlationship could be found in renal IL-18/IL-13 ratio. Conclusion It seems that the immune disturbance in systemic lupus erythematosus (SLE) can not be simply divided into Th1 predominant and Th2 predominant. It seems far complicated than this Th1/Th2 paradigm. It may be affected by the state of disease activity, the lesion location and the type of pathology.

OBJECTIVE To evaluate the roles of p15,p27 gene protein and expression in nasopharyngeal carcinoma(NPC).METHODS The EnVision immunohistochemical method was used to detect the expression of p15,p27 gene protein in nasopharyngeal carcinoma tissues of 43 NPC cases and non-tumor nasopharyngeal tissues of 21 cases.RESULTS ① The positive expression rates of p15,p27 gene protein were 65%,68% in NPC respectively.There were significant differences between NPC and non-tumor group(P0.05).③The positive expression of p15 gene protein was correlated to the positive expression of p27 gene protein(P

Objective To explore the function of p53 on regulating the expression of miR-148b in lung cancer cell line PC-9 and its corresponding molecular mechanism and the impact on cell proliferation. Methods Transient transfection of p53 eukaryotic expressing plasmids into lung cancer cell line PC-9 was performed to establish a cell model over-expressing p53. RT-PCR was used to explicit the impact of p53 on the expression of miR-148b. A reporter vector containing miR-148b promoter was used to investigate the function of p53 on regulating the transcription of miR-148b. Low-expressing miR-148b by transfecting its specific inhibitors , a CCK-8 assay was performed to explore the influence of miR-148b on the lung cancer cell proliferation inhibited by p53. Results Over-expression of p53 promoted miR-148b expression in lung cancer cell line PC-9. P53 could increase the luciferase activity driven by miR-148b promoters. Knockdown of miR-148b attenuated the impact of p53 on inhibiting the proliferation of PC-9 cells. Conclusion P53 inhibits the proliferation of lung cancer cell line PC-9 partially depending on miR-148b.

Objective To examine the complement component 1 Q subcomponent-binding protein (C1QBP) gene expression in human resistance choriocarcinoma cell lines and its parental cell line JeG-3,and to investigate whether silence C 1QBP by small interference RNA could reverse the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.Methods Expression of C1QBP mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR and western blot,respectively.The difference of C 1QBP expression was compared between human resistance choriocarcinoma cell lines and its parental cell line JeG-3.Sub-cellular location was proved by confocal immunofluorescence microscopy.A lentiviral vector containing short hairpin RNA (shRNA) targeting C 1QBP was constructed and cotransfected with the packaging plasmid mixture into 293T cells by lipofectamine 2000.The human resistance choriocarcinoma cell lines were infected with the packaged lentivirus.Real-time fluorogenic quantitative PCR and western blot were used to validate whether the C 1QBP gene expression was silenced.The cell counting kit 8(CCK8)was used to determine the drug sensitivity.Results (1)The C1QBP mRNA expression levels among four human resistance choriocarcinoma cell lines[JeG-3/floxuridiuum (FUDR),JeG-3/methotrexate (MTX),JeG-3/etoposide (VP),JeG-3/dactinomycin (KSM)] were 2.520±0.680,1.770±0.230,1.940±0.090 and 1.740±0.350 folds compared to that in JeG-3 cells.The C1QBP protein was higher expression level in human resistance choriocarcinoma cell lines than that in JeG-3.The immunofluorescence methods and confocal analysis showed that C1QBP localized predominantly in the mitochondrial matrix.(2)The C1QBP mRNA expression in JeG-3/FUDR cells after infected with lentiviral vector were decreased by 93.1％ (P＜0.01).The protein expression of C 1QBP in JeG-3/FUDR cells after infected with lentiviral vector were almost completely suppressed.The resistance indexes of four human resistance choriocarcinoma cell lines(JeG-3/FUDR,JeG-3/MTX,JeG-3/VP,JeG-3/KSM) were respectively 86.3％,93.9％,92.8％ and 89.9％,which were decreased remarkably by knockdown the C 1QBP expression (P＜0.05).Conclusions C1QBP is overexpressed in human resistance choriocarcinoma cell lines compared with parental cell line JeG-3.Inhibition of C 1QBP by lentivirus-mediated small interference RNA could effectively reverses the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.

BACKGROUND: Meseuchymal stem cells (MSCs) have extremely strong self-duplication ability and multidirectional ifferentiation potential. When bone marrow stromal cells (BMSC) are isolated and cultured in vitro, implanted in vivo, the distribution and colonization are still unclear, which is concerned with whether BMSC can be usedas target cells in clinic.OBJECTIVE: To explore the capacity of colonizing to the liver after allografting of green fluorescent protein (GFP) labeled MSCs of rats by different approaches.DESIGN: Factorial design.SETTING: Department of General Surgery, Second People's Hospital of Guangdong Province, Postdoctoral Workstation of Sun Yat-Sen University;Department of General Surgery, Zhujiang Hospital, Southern Medical University; Department of Organ Transplantation, Third Hospital Affiliated to Sun Yat-Sen University.MATERIALS: The experiment was performed at the Staff Room of Pharmacology, Basic Department, First Military Medical University of Chinese PLA from January 2003 to December 2004. A total of 36 clean adult SD rats were selected and randomly assigned into 5 groups: CCL4 plus portal vein transplantation group (n=6), portal vein transplantation control group (n=6), CCL4 plus caudal vein transplantation group (n=6), caudal vein transplantation control group (n=6) and mixed group (n=12).METHODS: ① MSCs were obtained from rat marrow and labeled with GFP. After amplifying in vitro, MSCs suspension was implanted with thin needle, with the volume of 0.5 mL/100 g. ②CCL4 plus portal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL4 2.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from portal vein. Portal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from portal vein. CCL4 plus caudal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL42.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from caudal vein. Caudal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from caudal vein. Mixed group: On the basis of the former 4 groups, 2 rats were implanted with non-labeled MSCs; Another 2 rats fed with CCL4 for 3 days and normal feed were established, without MSCs transplantation. ③At days 3 and 7 after transplantation expression of transplanted MSCs in liver of rats of each group were examined with fluorescent quantitative PCR.MAIN OUTCOME MEASURES: ①Results of MSCs isolation, purification, in vitro amplification and phenotype identification, ②result of GFP-labeled MSCs, ③observation of growth of rats following allografting of MSCs, and ④result of quantitative identification of GFP positive DNA amount in hepatic tissues of each group.RESULTS: Totally 36 experimental SD rats were involved in the result analysis. ①Percoll gradient separating medium was applied to isolate bone marrow of rats. The obtained cells were transferred and amplified,and then mostly showed coincident shuttle shape. Cells did not express CD34 and CD45, but CD29, CD44 and CD90 of MSCs, which were noncommitted stem cells in non-differentiating status that were different from hemopoietic stem cells in bone marrow. ②The green fluorescent cells appeared 24 hours after MSCs transfection. From hour 48 to 72 the number of positive cells significantly increased, with strong intensity.The transfection efficiency was 20%-30% under high-power field, and most of the cells were with green fluorescence. But green fluorescent cells did not appear in the MSCs cells as control. ③After allografting of labeled or non-labeled MSCs of rats with different approaches, at day 1 the rats were listless with bad food appetite, less mobilization; At day 2mostly of them had normal diet and mood, but there was no significant difference in rats of each group. ④The rats in each group with the exception of mixed group had green fluorescent protein positive cells in liver at days 3 and 7. The number of green fluorescent protein positive DNA was higher in liver tissues in the CCL4 plus portal vein transplantation group and CCL4 plus caudal vein transplantation group than in the portal vein transplantation control group and caudal vein transplantation control group (P＜0.05).CONCLUSION: Duration and amount of stem cells colonizing in liver may be associated with liver injury, while not related to the implantation approach. In normal animals with uninjured liver the stem cells can colonize in liver, and the amount is associated with transplantation approach and post-transplantation duration.

Objective To study the efficacy of laparoscopic tumor resection combined with iodine-125 and radiofrequency ablation in the treatment of rectal carcinoma with synchronous hepatic metastasis. Methods There were 30 patients diagnosed as rectal carcinoma with synchronous hepatic metastasis detected by CT scan. Hepatic metastases were confirmed by needle biopsy under laparoscopy. Laparoscopic radical resection of rectal carcinoma and metastatic hepatic tumors was performed. Those metastatic tumors that could not be resected were managed by RAF. Iodine-125 was planted in the tumors' site. Results Seven new hepatic metastases were found by the laparoscopic ulstrasound during the operation. 8 hepatic metastatic lesions were removed, 25 tumors located in the right liver were managed by RAF. All patients were followed-up from 12 to 25 months(average 22. 3 months), Local recurrence was found in 6 patients, the 1-year survival rate was 73% (22/30). Conclusions Laparoscopic excision, Iodine-125 and radiofrequency ablation in the treatment of rectal carcinoma with synchronous hepatic metastasis is safe、effective、minimally invasive.