Aim To investigate the effect of neferine on proliferation and invasion of human hepatocellular car-cinoma. Methods HepG2 and Bel-7402 cells were cultured in vitro with different concentrations of nefer-ine, then cell proliferation was observed by CCK-8 as-say; cell invasion was observed by transwell invasion assay; the protein expression of RhoA, RhoC and ROCK was detected by Western blot. Results CCK-8 results showed that neferine could significantly inhibit cell proliferation in a dose-dependent manner compared with the control group. Transwell invasion assay showed that cell invasion was significantly decreased with neferine 3 μmol · L-1 . Western blot results showed that RhoA, RhoC and ROCK protein expres-sion was decreased when neferine was co-incubated with hepatocellular carcinoma cells. Conclusion Nef-erine can inhibit proliferation and invasion of HepG2 and Bel-7402 cells, which is mediated mainly by the inhibition of RhoA, RhoC and ROCK protein expres-sion.

Objective To investigate the effects of targeted treatment of the carboplatin-Fe@C nanocage-loaded chitosan nanoparticles (C-Fe@CN-CN) combining external magnetic field on rats with transplanted liver cancer.Methods Twenty-four model rats with transplanted liver cancer were established and divided into four groups randomly (n =6).Abdominal exposure was carried out through a midline incision,and a cannula was inserted into the hepatic artery and fixed.Group A:saline water was injected as control,group B:saline water with 10 mg/kg free carboplatin was given,group C:saline water with C-Fe@CN-CN (equivalent dose of free carboplatin 10 mg/kg) was injected in absence of magnetic field,group D:saline water with C-Fe@CN-CN (equivalent dose of free carboplatin 10 mg/kg) was injected in presence of magnetic field for 30 min.All the animals were sacrificed and abdominal exposure was done again after 7 days.After tumors were reselcted,tumor weight and volume was measured,the inhibiting rate of tumor weight was calculated.Tumor and liver tissues were examined for histological changes.Results The growth of tumor was significantly inhibited after therapy with different forms of carboplatin.There was significant difference in the tumor weight of A,B,C,D groups [(0.85±0.12) g,(0.61±0.10) g,(0.48±0.09) g,(0.33±0.06) g,P ＜ 0.05,respectively].The inhibiting rates of tumor weight of B,C,D groups were 28.9 ％,43.4 ％,61.7 ％ respectively.The inhibiting rate of D group was highest which was 1.1 times higher than that of B group.There was also significant difference in the tumor volume of A,B,C,D groups [(1.06±0.24) cm3,(0.72±0.10) cm3,(0.50±0.07) cm3,(0.28±0.05) cm3,P ＜ 0.05,respectively].The tumor volume of group A was largest which was 2.8 times larger than that of group D.In group D,tumor tissues from six rats presented severe necrosis,and nanoparticles were concentrated in the necrotic tissue.In group C,five rats presented middle necrosis,one rats presented severe necrosis.There was no concentration of nanoparticles in the necrotic tissue.In group B,four rats presented middle necrosis,two rats presented mild necrosis.In group A,six rats presented mild necrosis.Conclusion C-Fe@CN-C can significantly increase the therapeutic effects of carboplatin by hepatic artery injection combining with an external magnetic field on the tumor.

Objective To investigate the immune regulatory effects of allogeneic bone marrowderived mesenchyrnal stem cells (MSCs) co-transplanted with islets. Methods The 18 diabetic mice were randomly divided into 3 groups: Diabetic group, without any transplantation; Islet transplantation group, in a sterile operation, with 10 μl purified islets (about 200 islets)transplantation to the left renal subcapsule of recipients; Islet + MSCs transplantation group, in addition to transplantation as the former group, 1 × 106 MSCs were given to the recipients via tail vein on 3, 2 and 0 days before islet transplantation. Blood glucose in recipient mice was monitored for 30consecutive days after transplantation. Pathological characteristics of left kidney were analyzed on the day 14 and 28 after transplantation. Th1/Th2, Tc1/Tc2, naive and memory T cells from peripheral blood and bone marrow-derived dendritic cells (DCs) were analyzed by multi-color flow cytometry.Results As compared with the islet transplantation group, blood glucose was significantly reduced,inflammatory cell infiltration decreased in the place of transplantation, graft survival prolonged, the number of Th1 and Tc1 cells was obviously reduced, the number of Th2 and Tc2 cells increased, the ratio of Th1/Th2 and Tc1/Tc2 cells was significantly decreased, the naive and memory T cells were significantly inhibited, the maturity of DCs and the secretion of interleukin-12 decreased in the islet transplantation group. Conclusion Through the immunomodulation on T-cell and DCs function,MSCs can alleviate graft verse host disease and prolong allograft survival.

Objective Compare the reproductive activity of pancreatic stem cells isolated from diabetic rat with that of normal rat to assess the effect of microenvirement on stem cells'proliferation.Methods Diabetic rat model were prepared by peritoneal injecting 60mg/kg streptozotocin.Pancreata from 10 diabetic rats and 10 normal rats were trimmed and digested into single cells which were then inoculated onto the culture plate respectively.After passaging 3 generation the cells were identified by nestin immunostaining.During culturing the cell growth curve was observed and the cell cycle of the third and seventh generation cultured cells was evaluated by flow cytometry.Results After passaging pancreatic stem cells was purified and identified by nestin immunostaining.The reproductive activity of diabetic rat was much better[ratio of s stage:(20.7?2.7)% vs(14.1?2.5)%,P0.05].Conclusions The stem cells are stimulated to proliferate after islet injure.The reproductive activity of cells can be provoked by the microenvironment.

Objective To explore the feasibility of la pa roscopic total extraperitoneal hernia repair without the use of the balloon diss ector. Methods Laparoscopic total extraperitoneal hernia repai r using no balloon dissector was completed in 10 cases of groin hernia from Nove mber 2003 to April 2004, including 7 cases of indirect inguinal hernia, 1 case o f occult indirect hernia combined with direct inguinal hernia, and 3 cases of di rect inguinal hernia (1 of which was recurrent hernia). Results All the 10 operations were successful. The mean operation time was 69.5 min, t he mean blood loss was 11 ml, the mean postoperative hospital stay was 2.1 days , and the mean duration to normal activities, 5.2 days.Complications included 1 case of disassociated inferior epigastric artery, 1 case of operation in wrong anatomic layer, and 1 case of laceration of the peritoneum, respectively. Follow -up observations for 1~6 months found no recurrence and neuropathic pain. Conclusions Laparoscopic total extraperitoneal hernia repair without the use of the balloon dissector is feasible, safe and effective.

OBJECTIVETo investigate the effects of all-trans-retinoic acid(ATRA) and polyphenon-100 (PP) on genetic instability of human tumor cells via their role in alteration of microsatellite sequence(MS) and the expression of mismatch repair gene hMLH(1) and hMSH(2) in RER(+) (replication error) cells.

METHODSRER(+) colon cancer cell line was used as a host for lipofection with pCMV-CAR in which a foreign (CA)(14) repeat was inserted in the coding sequence of LacZ reporter gene, resulting in misreading LacZ frame. Any mutation which made the base number of (CA)(14) tract to be 3-fold resumed normal reading frame of LacZ, and thus led to expression of beta-galactosidase. Variable expression of LacZ in the transfectant cells resulting from RATA or PP treatment was measured by OD reading at lambda 620 after X-gal staining. Expression of mismatch repair genes of hMLH(1) and hMSH(2) was examined at mRNA level by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSATRA at 1 mu mol/L, 0.1 u mol/L and PP at 3 mu g/ml had no significant inhibitory effect on cell proliferation. After being treated with ATRA or PP for 1 week, the blue cells of RKO transfectant clones were significantly reduced, and this meant the mutation of exogenous (CA)(14) in RKO cells were inhibited. But no expression of hMLH(1) and hMSH(2) was observed.

CONCLUSIONThe above data showed both ATRA and PP had inhibitory effects on MS instability of cancer and thus demonstrated directly their beneficial role in stabilization of genomic DNA. However, the present authors have not observed any expression of hMLH(1) and hMSH(2) in RKO cells treated with ATRA or PP.

Adaptor Proteins, Signal Transducing ; Carrier Proteins ; Catechin ; pharmacology ; Cell Division ; drug effects ; Colonic Neoplasms ; genetics ; pathology ; DNA-Binding Proteins ; Gene Expression ; drug effects ; Humans ; Lac Operon ; genetics ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; Mutation ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Plasmids ; genetics ; Proto-Oncogene Proteins ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Transfection ; Tretinoin ; pharmacology ; Tumor Cells, Cultured ; drug effects

Objective To discuss the feasibility of laparoscopic closure of the internal ring and high ligation of the hernial sac in children with indirect inguinal hernia.Methods Laparoscopic closure of the internal ring and high ligation of the hernial sac was conducted in 21 children with indirect inguinal from January 2003 to December 2004 in this hospital.A 5-mm 30? laparoscope was introduced through the umbilicus.The internal ring and the posterior crus(PC) on the side of hernia were identified.A 3-mm dissection forceps was placed on the midline between the umbilicus and pubic symphysis.At the skin projection of the internal ring on the side of hernia,a round-tip needle with 3-0 Prolene suture grasped by a regular needle holder was passed directly into the abdominal cavity to suture the posterior crus and then penetrate out of the abdomen via the roof of the internal ring through the musculoaponeurotic arch of the transversus abdominis.Then the Prolene suture was cut off,and passed by using the Endoclose device into the extraperitoneal cavity at the skin projection of the internal ring.The dissection was continued from without outward around the sac to encircle the neck and an extracorporeal high circumferential ligation of the sac was finished.Results The operation was successfully completed in all the 21 cases.The operating time was 16~50 min(mean,32.9 min),the postoperative duration in hospital was 1~2 d,and the time to normal activity,1~4 d(mean,1.4 d),respectively.No postoperative bleeding,hematoma of scrotum,or incisional infection occurred.Follow-up checkups in the 21 cases for 1~24 months(mean,15 months) showed no recurrence.Conclusions Laparoscopic closure of the internal ring and high ligation of the hernial sac is a novel,feasible,reliable and effective herniorrhaphy for the treatment of pediatric indirect inguinal hernia.

Objective To evaluate the dissection and hemostasis effects of harmonic scalpel in laparoscopic surgery. Methods The effect in patients underwent laparoscopic surgery with harmonic scalpel or with high frequency electricity scalpel was compared. Results All the operations were done by laparoscopy.No complications or massive blood loss occurred during and after operation.The average operation time ,blood loss of operation with harmonic scalpel were less than that with high frequency electricity scalpel. Conclusions Harmonic scalpel has precise cutting , controlled coagulation, and less tissue injury,it can increase the safety and microsurgery techinque of laparoscopic surgery.It is a very important equipmeat and useful for laparoscopic surgery .

Objective To compare the postoperative changes in immune function of patients undergoing robot-assisted laparoscopic cholecystectomy(RLC) and conventional laparoscopic cholecystectomy(CLC).Methods The clinical data of 52 cases of gallbladder stones and gallbladder polyps treated with RLC(23 cases),and with LC(29 cases) was compared.Blood samples from both groups were taken to measure immune markers(T cells,IgA,IgM,IgG,C3,C4) before operation and in the first postoperative morning.Results The postoperative levels of CD4/CD8 and IgM decreased from 1.56?0.67,1.42 ?0.58 to 1.30?0.53,1.26?0.46 respectively after RLC(all P0.05).Conclusions RLC and CLC showed no significant postoperative effect on cellular immune function,but both had some effect on humoral immune function.

Objective To study a new method for internal drainage and primary suture after laparoscopic choledocholithotomy (LCH). Methods Twenty-five patients with chronic cholecystitis combined with cholecystolithiasis and choledocholithiasis were performed laparoscopic cholecystectomy and choledocholithotomy and extraction of stones with choledoscopy. Results One case of Mirizzi' syndrome converted into laporotomy and 24 cases were successfully performed. Excluding neither internal nor external drainage was carried out with primary closure of common bile duct (CBD) in 1 case and T-tube were placed in 4 cases, the made-self 3mm～5mm diameter silica tubes with one-way hook were placed into the common bile duct for internally drainage in 19 cases. The time of operation was from 90 to 420 minutes with a mean of 145 minutes. 1～5 calculus were removed from CBD. A mean of hospitalized days stay was 7.5 day. Conclusions The operation of internal-tube drainage of LCH frees either the inevitably risk of hyper pressure of biliary duct because of the spasm of Oddi's sphincter or disadvantage of T-tube for drainage after LCH, which is new method both safety and practicality, both simpleness and easiness, and accords with the physiological needs of the human body.