Construction and identification of mouse Krüppel-like factor 4 recombinant lentiviral expression vector and its effect on proliferation of RAW264.7 cells / 天津医药

Objective To construct and identify a lentiviral vector carrying mouse Krüppel-like factor 4 (KLF4) gene, and establish RAW264.7 cell line of peritoneal macrophages that over-expressed KLF4. Methods KLF4 gene was cloned using the measure of polymerase chain reaction (PCR). Then the recombinant transfer vector pLVX-KLF4 (pLVX-KLF4-mCMV-ZsGreen-PGK-Puro) was constructed. The pLVX-KLF4 was confirmed through PCR, restriction enzyme digestion and sequencing. The correct recombinant transfer vector together with its two helper virus vectors (psPAX2 and pMD2.G) were cotransfected into the 293T cells by Lipofectamine? 3000. The supernatant of 293T was harvested to infect RAW264.7 cells. Flow cytometry (FCM) was used to test the viral titer of the expression level of green fluorescent protein. The expression of KLF4 mRNA in RAW264.7 cells was measured by real-time PCR. Results The restriction enzyme digestion, PCR and sequencing confirmed that the transfer lentiviral vector pLVX-KLF4 was constructed successfully. KLF4 mRNA was over-expressed in Lenti-KLF4 transfected RAW264.7 cells than that of wild type RAW264.7 cells (P<0.05). In transfected RAW264.7 cells, KLF4 mRNA was over-expressed (P<0.05). The recombinant lentivirus of KLF4(Lenti-KLF4)titer was 2.05×108 TU/mL measured by FCM.The flow cytometry results showed that the S phase fraction was prolonged and G0/G1 was arrested in the over-expressed KLF4 of RAW264.7 cells. The EdU showed that the up-regulated expression of KLF4 gene stimulated the proliferation of RAW264.7 cells. Conclusion The recombinant lentiviral vector, which can effectively express KLF4 mRNA, has been successfully constructed. The up-regulated KLF4 gene may increase the proliferation of RAW264.7 cells.

Simultaneous laparoscopic excision for rectal carcinoma and synchronous hepatic metastasis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Rectal carcinoma patients are often accompanied by hepatic metastasis. The aim of this study was to evaluate the therapeutic efficacy of simultaneous laparoscopic excision for rectal carcinoma with synchronous hepatic metastasis.</p><p><b>METHODS</b>A total of 41 patients with rectal carcinoma and synchronous hepatic metastasis detected by CT scan were included in this study. Among them, 23 patients underwent laparoscopic surgery and 18 patients underwent traditional open surgery to simultaneously remove the rectal tumor and hepatic metastasis lesions. All patients received postoperative adjuvant chemotherapy. All the patients were followed up from 36 to 72 months (mean 45.3 months).</p><p><b>RESULTS</b>All the operations were performed successfully and no patient was turned to open surgery in laparoscopic group. The mean blood loss, the mean postoperative hospital stay, the mean blood transfusion and the mean intestinal functional recovery time showed a significant difference between the two groups (P < 0.05). The 1-, 3- and 5-year survival rates were 82.6%, 43.5% and 8.6% in the laparoscopic group, without significant difference compared with the open group (77.8%, 38.9% and 0) (P > 0.05).</p><p><b>CONCLUSIONS</b>Simultaneous laparoscopic excision for rectal carcinoma and synchronous hepatic metastasis is safe and effective with similar survival achieved by the traditional open abdominal surgery.</p>

Signal transduction pathway in liver fibrosis regulated by human hepatocyte growth factor / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the changes in the gene expression profiles in HepG2 cells transfected by human hepatocyte growth factor (hHGF) and analyze the signal transduction pathway in liver fibrosis regulated by hHGF.</p><p><b>METHOD</b>A 20,000 gene cDNA microarray (Affymetrix) was used to examine the gene expressions in the HepG2 cells transfected by hHGF. The differentially expressed genes were identified and some genes with possible contribution to hepatic fibrosis were subjected to real-time PCR analysis.</p><p><b>RESULT</b>The differentially expressed genes were mostly transcription regulatory molecules, cytokines, signal transduction, glucose metabolism, lipid metabolism. The results of real-time PCR showed up-regulated STAT1 and MAPK1 expression in the cells as were consistent with genechip analysis results.</p><p><b>CONCLUSION</b>hHGF gene transfection results in the gene expression profile changes in HepG2 cells. HGF may regulate liver fibrosis via the JAK/STAT and MAPK pathways.</p>

Immunological influence of iodine-125 implantation in patients with hepatocellular carcinoma resection / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of iodine-125 implantation on immune cell subsets and cytokine production in patients undergoing hepatocellular carcinoma (HCC) resection.</p><p><b>METHODS</b>Sixty-eight patients with resectable HCC were randomly divided into intrahepatic iodine-125 embedding group and control group. The percentages of T lymphocyte phenotypes (CD3(+), CD4(+), CD8(+)), NK cells, and the plasma concentrations of IL-4, IL-10, IL-12 and IFN-gamma of the patients were determined with flow cytometry and ELISA, respectively.</p><p><b>RESULTS</b>In the control group, the postoperative CD3(+) and CD4(+) immunocytes were (39.38-/+6.98)% and (24.34-/+3.18)%, significantly lower than the preoperative levels [(62.58-/+8.67)% and (30.63-/+4.19)%, respectively, P<0.05)]. The postoperative concentrations of IL-4 and IL-10 were (29.83-/+7.99) and (87.54-/+2.89) ng/L, significantly higher than the preoperative levels (10.35-/+8.76 and 64.25-/+2.54 ng/L, respectively, P<0.05). In the treatment group, the percentages of the immunocytes and cytokine concentrations underwent no significant changes after the operation, but postoperative IL-12 (89.46-/+11.43 ng/L) and IFN-gamma (47.78-/+5.45 ng/L) levels were significantly higher than the preoperative levels (36.13-/+9.16 and 7.14-/+2.17 ng/L, respectively, P<0.05). Significant differences were found between the two groups in the postoperative CD3(+) and CD4(+) immunocytes and IL-4, IL-10, IL-12 and IFN-gamma levels.</p><p><b>CONCLUSION</b>Iodine-125 implantation can strongly stimulates the anti-tumor immune response in HCC patients by increasing CD3(+) and CD4(+) immunocytes and promoting Th2/Th1 deviation.</p>

Applied anatomy of the inguinal region and laparoscopic inguinal hernia repair / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the regional anatomy of the inguinal region and abdominal ring and provide accurate anatomic basis for the clinical application of total peritoneum intraperitoneal onlay mesh (TPIPOM).</p><p><b>METHODS</b>The structures of the inguinal region and those surrounding the abdominal ring of 18 cadavers (11 males and 7 females, 36 sides) were dissected to measure the lengths from the nerves and vessels to the corresponding anatomical landmarks.</p><p><b>RESULTS</b>The average distances from the point where the iliohypogastric nerve (IHN) ran through the obliquus internus abdominis to anterosuperior iliac spine and from the point where the IHN ran through the aponeurosis of the external oblique muscle of the abdomen to the superior margin of the symphysis pubica were 4.10±0.89 cm and 5.02±1.46 cm, respectively. The average distances from the point where the ilioinguinal nerve (IGN) ran through the obliquus internus abdominis to the anterosuperior iliac spine and from the point where IGN ran through the aponeurosis of the external oblique muscle of the abdomen to the superior surface of the tuberculum pubicum were 3.09±0.81 cm and 3.84±0.89 cm, respectively. We established the regional anatomy model of the structures surrounding the abdominal ring.</p><p><b>CONCLUSION</b>The quantitative measurement of important structures of the inguinal region and establishment of the regional anatomy model of the surrounding structures of the abdominal ring can provide a valuable reference to reduce intraoperative and postoperative complications of TPIPOM.</p>

Effect of ROUX-en-Y anastomosis and Billroth-II anastomosis after subtotal gastrectomy on blood glucose in type 2 diabetic patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of ROUX-en-Y anastomosis and Billroth-II anastomosis after subtotal gastrectomy on blood glucose in patients with type 2 diabetes.</p><p><b>METHODS</b>From January 2006 to December 2009, 26 and 24 type 2 diabetic patients underwent subtotal gastrectomy for gastric cancer or gastric ulcer and received subsequent ROUX-en-Y anastomosis and Billroth-II anastomosis, respectively. Perioperative and postoperative blood glucose and glycosylated hemoglobin (HbA1c) were detected in these patients to identify the variations.</p><p><b>RESULTS</b>In ROUX-en-Y group, the fasting blood glucose (FBG) of the patients decreased significantly in the first month after the operation (t=4.46, P<0.05), and the 2-hour postprandial glucose (2hPG) and HbA1c also underwent significant reductions in the first postoperative week and month, respectively (t=3.5, P<0.05; t=2.21, P<0.05). In Billroth-II group, the FBG decreased significantly till 6 months after the operation (t=2.0, P<0.05), and HbA1c reduction occurred 3 months after the operation (t=2.61, P<0.05). There were significant differences in FBG in the first postoperative month, 2hPG in the 6th month, and HbA1c in the 3rd month between the two groups (P<0.05).</p><p><b>CONCLUSION</b>The ROUX-en-Y anastomosis and Billroth-II anastomosis after subtotal gastrectomy can both reduce blood glucose of type 2 diabetic patients, but the former approach has better effect.</p>

Laparoscopy combined with endoscopic and digital subtraction angiography for treatment of severe acute pancreatitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of laparoscopy combined with endoscopic and digital subtraction angiography in the treatment of severe acute pancreatitis (SAP).</p><p><b>METHODS</b>Nine-seven SAP patients were randomly divided into group A (n=32) with conventional treatment and group B (n=35) with combined treatment with laparoscopy, endoscopic and digital subtraction angiography in addition to the conventional treatment. The clinical indices and therapeutic effects in the 2 groups were compared.</p><p><b>RESULTS</b>After treatment, the patients in group B had significantly lower APACHE score than those in group A (P<0.05), with also better hepatic, renal and lung functions (P<0.05). The serum TNF-alpha and IL-1beta levels was significantly lower, but IL-10 significantly higher in group B (P<0.05). The CT SPN was significantly lowered in group B as compared with that in group A (P<0.05). The patients in group B had significantly lower rate of organ failure (P<0.01) and higher successful rate in organ failure management (P<0.05), with also lowered inhospital mortality than those in group A (P<0.05).</p><p><b>CONCLUSION</b>Laparoscopy combined with endoscopic pancreaticobiliary duct drainage and digital subtraction angiography in addition to conventional treatment significantly improves the outcome and decreases the mortality of SAP patients.</p>

Simultaneous laparoscopic excision for the treatment of rectal carcinoma and the synchronous hepatic metastasis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic efficacy of simultaneous laparoscopic excision for the treatment of rectal carcinoma and synchronous hepatic metastasis.</p><p><b>METHODS</b>Totally 38 patients with rectal carcinoma and synchronous hepatic metastasis detected by CT scan were included in this study. Among them, 23 patients in the group A were treated with laparoscopic surgery, and the other 18 patients in the group B with traditional abdominal operation to resect the rectal tumor and hepatic metastasis simultaneously. All patients received postoperative chemotherapy.</p><p><b>RESULTS</b>All the patients were treated successfully with no postoperative death in both groups. The mean operative time was 350 +/- 45 min in group A versus 342 +/- 38 min in group B (P > 0.05). The mean blood loss was 275 +/- 96 ml in group A versus 590 +/- 85 ml in group B (P < 0.01), and the average hospital stay was 12 +/- 1.5 days in group A versus 16 +/- 2.5 days in group B (P < 0.05). Only one patient in group A received blood transfusion of 200 ml during operation, while the average blood transfusion in group B was 500 +/- 100 ml (P < 0.01). The follow-up duration was from 36 to 72 months with an average duration of 45.3 months. The 1-, 3- and 5-year survival rates were 82.6%, 43.5% and 8.6% in the group A, versus 77.8%, 38.9% and 0% in group B, respectively (P > 0.05).</p><p><b>CONCLUSION</b>Simultaneous laparoscopic excision of rectal carcinoma and synchronous hepatic metastasis is safe, effective and minimally invasive with a similar survival achieved by traditional open abdominal operation.</p>

Laparoscopic placement of total peritoneum intraperitoneal onlay mesh in patients with inguinal hernia / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the efficacy of laparoscopic placement of total peritoneum intraperitoneal onlay mesh (TPIPOM) for treatment of inguinal hernia.</p><p><b>METHODS</b>TPIPOM was placed laparoscopically in 125 cases of inguinal hernia, and the clinical outcomes of the patients were observed and compared with 64 patients receiving transabdominal preperitoneal laparoscopic mesh repair (TAPP) and 53 with total extraperitoneal laparoscopic hernioplasty (TEP).</p><p><b>RESULTS</b>The laparoscopic operations were successfully performed in all the patients. In TPIPOM, TAPP and TEP groups, the operating time was 30.8-/+10.3 min, 68.4-/+22.4 min and 69.5-/+23.4 min (P<0.05), the mean hospital stay was 3.8-/+1.3 days, 4.3-/+1.5 days and 4.5-/+1.6 days (P<0.05), the average time to ambulation was 1.2-/+0.5 days, 1.8-/+0.7 days and 2.2-/+0.8 days (P<0.05), the duration of pain was 1.0-/+0.5 days, 1.6-/+0.9 days and 1.9-/+0.8 days (P<0.05), and the cost was 5000.8-/+800.5 yuan, 8000.5-/+950.6 yuan and 8900.2-/+750.3 yuan (P<0.05), respectively. No scrotum edema occurred in these patients. The patients were followed up for 59.9-/+6.5 months and recurrence was found.</p><p><b>CONCLUSION</b>TPIPOM is safe and effective for management of inguinal hernia with such advantages as minimal invasion, simple procedures, shorter operation time, reduced relapse and quick recovery.</p>

Intraoperative radiofrequency ablation and 125I therapy for preventing local recurrence in hepatocellular carcinoma after hepatectorny / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of intraoperative radiofrequency ablation and 125I therapy for preventing local recurrence in hepatocellular carcinoma (HCC) after hepatectomy.</p><p><b>METHODS</b>Seventy-eight HCC patients with a tumor close to the first or the second hepatic portal underwent hepatectomy with a resection margin less than 1 cm from 1999 to 2001. All patients were randomly divided into control group and combined treatment group according to their check-in date (odd or even). In the control group, 38 patients were treated with hepatectomy alone, whereas in the combined group, 40 patients were treated with hepatectomy plus radiofrequency ablation and 125I implantation on surgical margin. All patients were followed up regularly.</p><p><b>RESULTS</b>the 1-, 3- and 5-year recurrent rate was 7.5%, 30.0% and 45.0% in the combined treatment group versus 18.4%, 60.5% and 86.8% in the control group, respectively, with a significant difference in the 3-year (chi2 = 7.340, P < 0.01) and 5-year recurrent rates (chi2 = 15.740, P < 0.01). The 1-, 3- and 5-year survival rate was 92.5%, 67.5% and 30.0% in the combined group versus 81.5%, 39.4% and 18.4%, respectively.</p><p><b>CONCLUSION</b>Intraoperative radiofrequency ablation and 125I therapy on the resection margin is effective to prevent local recurrence in HCC patients after hepatectomy, and to prolong their postoperative tumor free survival.</p>

Laparoscopy and digital subtraction angiography in the treatment of severe acute pancreatitis / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effect of laparoscopy and digital subtraction angiography in the treatment of severe acute pancreatitis (SAP).</p><p><b>METHODS</b>Seventy-five SAP patients were randomly divided into tow groups: (1) Conventional treatment group (group A, n = 35); (2) Conventional treatment combined with laparoscopy and digital subtraction angiography treatment group (group B, n = 40). The clinical parameters and treatment results in the 2 groups were compared.</p><p><b>RESULTS</b>After treatment, APACHE II score in group B was significantly lower than that in group A (P < 0.05). The functions of liver, renal and lung were recovered (P < 0.05, respectively), levels of serum TNF-alpha and IL-1 beta were significantly lower, but IL-10 significantly elevated (P < 0.05), the CT SPN was much lower (P < 0.05); rate of organ failure significantly decreased (P < 0.01), the successful rate of organ failure treatment was increased (P < 0.05); the in hospital mortality decreased (all P < 0.05).</p><p><b>CONCLUSIONS</b>Laparoscopy and digital subtraction angiography in combination with conventional treatment for SAP significantly improves the outcome and decrease the mortality of SAP.</p>

Endoscopic thyroidectomy: a comparison of the trans-axilloareolar approach and the trans-thoracoareolar approach / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the advantages and shortcomings of endoscopic thyroid surgery by trans-axilloareolar approach and trans-thoracoareolar approach.</p><p><b>METHODS</b>Endoscopic thyroidectomy was performed in 238 cases from December 2003 to January 2006. The patients were randomly divided into two groups. One hundred and twenty-three patients received the trans-axilloareolar approach and 115 patients randomly received the trans-thoracoareolar approach procedure.</p><p><b>RESULTS</b>All the operations were succeed. The operating duration of trans-axilloareolar approach group and trans-thoracoareolar approach group were (69 +/- 29) min and (70 +/- 25) min (P > 0.05), the blood lost were (38 +/- ll) ml and (40 +/- 13) ml (P > 0.05), the average hospitalized days were (4 +/- 1.3) d and (4.5 +/- 1.2) d (P > 0.05), the rate of satisfaction with the cosmetic effects of the procedures were 97.5% and 85.2% (P <0. 05). There were no conversions to open surgery or any complications. The drainage was removed at 24 h to 36 h after the operation.</p><p><b>CONCLUSIONS</b>Endoscopic thyroidectomy through trans-axilloareolar approach and trans-thoracoareolar approach is feasible, safe and cosmetic. The cosmetic effects of the trans-axilloareolar approach is better than the trans-thoracoareolar approach. The procedure selection depends on both the patient request and the technology of the surgeons.</p>

Laparoscopic high ligation of hernia sac and median umbilical fold covering internal ring in treating pediatric indirect inguinal hernia / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect and clinic practice of laparoscopic high ligation of hernia sac and median umbilicus fold covering internal ring in pediatric indirect inguinal hernia.</p><p><b>METHODS</b>Five hundred and fifty-two children with indirect inguinal hernia were randomly divided into group A (control group) and group B (combination group) according to their check-in dates (odd or even). In group A, 275 children were treated with laparoscopic high ligation of hernia sac. In group B, 277 children were treated with laparoscopic high ligation of hernia sac and suturing the median umbilical fold to internal ring.</p><p><b>RESULTS</b>The operation was successfully performed in all 552 patients. The operating time in group A and B were (12.3 +/- 5.5) min and (20.6 +/- 4.2) min (P<0.05) respectively; and the hospital stay were (2.2 +/- 1.5) d and (2.4 +/- 1.3) d (P>0.05); the day of restoring normal active were (1.2 +/- 0.5) d and (1.3 +/- 0.6) d (P>0.05); the recurrence rate were 2.2% (6/275) and 0 (0/277) (P<0.05). No complications, such as abdominal cavity infection and scrotum hematoma occurred during the follow-up of 22 months to 55 months.</p><p><b>CONCLUSIONS</b>Laparoscopic high ligation of hernia sac and median umbilical fold covering internal ring is safe and effective in treating indirect inguinal hernia of the children.</p>