OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

Objective To probe into the clinical curative effect of laparoscopic high ligation of spermatic vein in the treatment of critical varicocele.Methods The clinical data of the 26 eases of laparoscopic high ligation of apermatic vein in the treatment of critical varicocde were reviewed and analyzed in the last two years in this hospi- tal.Results All the 26 cases had been conducted smoothly with the operation time 25～50min,an average of(28?3)min.After 3～24 months being followed-up,all the symptoms and signs disappeared with no relapse and testicle a- trophy.Eight wives of the patients who had been operated on got pregnant.Conclusion With soon recovery and small wound,it was safe to adopt laparoscopic high ligation of spermatic vein in the treatment of critical varicocele, especially for critical two-sided varicocde.

OBJECTIVETo construct one lentiviral vector containing mouse SRY-related high mobility group-box gene 9 (SOX9) and transfect the murine bone mesenchymal stem cells (mBMSCs) in vitro and observe the expression of target gene.

METHODSRNA from the vectors containing mouse SOX9 gene were extracted and SOX9 genes were amplified by reverse transcription-Polymerase Chain Reaction (RT-PCR). The SOX9 genes were connected into lentiviral vectors pGC-FU. Then pGC-FU-SOX9 transduced into 293T cells to produce recombinant lentivirus called as Lenti-SOX9-EGFP. mBMSCs were transfected. The expression of target gene was detected by immunofluorescence, RT-PCR and Western Blot.

RESULTSLenti-SOX9-EGFP was recombined successfully and transduced efficiently into mBMSCs. The expression of SOX9 gene was confirmed by RT-PCR and Western Blot.

CONCLUSIONLentiviral vector of mouse SOX9 gene can transfect successfully into mBMSCs. Meanwhile, SOX9 gene may be expressed in mBMSCs. This will provide the target cells for the following study about SOX9 gene repairing cartilage injury.

Animals ; Female ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Mesenchymal Stromal Cells ; metabolism ; Mice ; Osteoarthritis ; therapy ; Reverse Transcriptase Polymerase Chain Reaction ; SOX9 Transcription Factor ; genetics ; Transduction, Genetic ; Transfection

Objective:To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for re?pairing articular cartilage injury in vivo. Methods:Rabbit bone marrow mesenchymal stem cells(BMSCs)were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treat?ment. After animals anesthesia,a full thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile,the transfected cells were implanted to repair the rabbit model with full thickness cartilage defects. Cartilage defects tissue was observed with light microscope,electron microscope,HE and im?munohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation. Re?sults:At 3 days after the transfection,Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection,the expression of collagen typeⅡbegan and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox 9 gene. Histologi?cal observation showed that at 6 weeks after the operation,the defects in the experimental group was filled with hyaline like cartilage tissue,12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively,the defects were filled with fibrous tissues in control groups. Meanwhile,immunohistochemical stain?ing of sections with typeⅡcollagen antibodies showed the proteins in the regenerated tissue stained positive for typeⅡcolla?gen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances. Conclusion:Overexpression of Sox9 gene on rab? bit bone marrow mesenchymal stem cells(BMSCs)promote the repair of cartilage defect.

OBJECTIVETo summarize and compare the short-term and long-term clinical efficacy of percutaneous transhepatic biliary drainage (PTBD) and percutaneous transhepatic biliary stent (PTBS) in the treatment of malignant obstructive jaundice.

METHODS210 cases of malignant obstructive jaundice underwent interventional therapy, of which 161 cases of drainage catheters placement and 49 cases of metallic stent implantation. Follow-up information was obtained through telephone review or check-up records.

RESULTSThe technical success rate of technique was 100%. At 3 - 5 days after treatment, the serum total bilirubin in 15 metallic stent-treated patients was decreased by (178.04 +/- 42.32) micromol/L, and direct bilirubin by (83.97 +/- 23.63) micromol/L. Compared with those of 28 cases treated with drainage catheters: (95.67 +/- 34.28) micromol/L and (49.84 +/- 28.21) micromol/L, there were statistically significant differences between the two groups (P = 0.017 and P = 0.035). At 6 - 9 days after treatment, the serum total bilirubin in 28 cases of metallic stent group was decreased by (188.22 +/- 79.90) micromol/L, and that in 126 cases of drainage catheter group decreased by (141.39 +/- 65.32) micromol/L. The difference was statistically significant (P = 0.014). But the decline value of direct bilirubin had no significant difference. The median patency period and the median survival time of the drainage catheter group were 60 and 148 days, respectively, those of metallic stent group were 197 days and 245 days. There were statistically significant differences between the two groups (P < 0.05).

CONCLUSIONThe results of this study indicate that the short-term and long-term efficacies of metallic stent implantation are better than those of catheter drainage technique.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; complications ; Bilirubin ; blood ; Drainage ; instrumentation ; methods ; Female ; Gallbladder Neoplasms ; complications ; Humans ; Jaundice, Obstructive ; blood ; etiology ; therapy ; Liver Neoplasms ; complications ; Male ; Middle Aged ; Pancreatic Neoplasms ; complications ; Stents ; Survival Rate

Contributions of Xin'an medical school and physicians to acupuncture theory were introduced in the article. Academic theories or characteristics of several physicians of Xin'an school such as YANG Xuan-cao, WU Kun, WANG Ji, WU Yi-ding, ZHENG Mei-jian and XU Chun-fu, et al were sorted out. Contributions of inheriting and illustrations on acupuncture theory were analyzed so as to expound its significance and value on modern acupucture clinic.
Acupuncture ; education ; history ; manpower ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Physicians ; history ; Schools, Medical ; history ; manpower

Objective To study the impacts of the Three Gorges dam and change of water level on the survival of the local rodents, and to provide scientific basis to control the outbreak of rodent-borne diseases.Methods Four villages located around the Three Gorges dam were selected in the study. The mouse populations by using Elton night trapping method was monitored. Metallic spring traps were set for two consecutive nights. The mouse density and identified the mouse species was calculated. The mouse species indoor and outdoor, as well as the mouse density indoor and outdoor were compared. The impacts of water level in the dam and cleaning work on local mouse density were also analyzed. Results A total of 678 mice were caught in this study, 517 were caught indoor and 161 outdoor. Indoor dominant species was flavipectus; accounting for 36.49%(189/517), while outdoor was apodemus, reaching 56.88% (91/161). For mouse species, there was a significant difference between indoor and outdoor(x2 = 678.00, P ＜ 0.01 ). The average mouse density was 8.44%(678/8036) in trap nights. Indoor mouse density reached 14.44%(517/3581 ), which was significantly higher than that of outdoor(3.61%, 161/4455 ).For mouse density, there was a significant difference between indoor and outdoor(x2 = 301.04, P ＜ 0.01 ). When the water level was up to 156 m, mouse density reached 10%(513/5132), which was higher than that of before (5.68%, 165/2904). There was a significant difference in mouse density before and after reserving water (x2 = 44.68, P ＜ 0.01 ). With the change of water level, upstream mouse density formed a high platform from May 2007 to May 2008, followed by 12.25%(80/653), 13.16%(90/684), 12.95%(90/695), and decreased to 8.38%(28/334) after cleaning of the dam. Conclusions The Three Gorges dam and change of water level actually alter the survival environment of the local mouse, and affect local mouse density and mouse species. These may lead to local outbreak or epidemic of rodent-borne diseases.

OBJECTIVETo study the regularity of splanchnic hemodynamic changes after orthotopic liver transplantation (OLT) for patients with portal hypertension. At the same time, effect of such changes on splenomegaly, hypersplenism, collateral circulation and the postoperative liver function was discussed.

METHODSBetween June 2002 and October 2005, 173 liver transplantations were performed. In 38 patients with portal hypertension undergoing OLT, the following parameters were measured before surgery and subsequently at 1, 3, 5, 7 days, 1, 6 months and 1, 2, 3 years after operation by using Color Doppler sonography: portal blood flow mean velocity (PBV), portal blood flow volume (PBF), hepatic artery resistance indexes (HA-RI) and spleen size. The same parameters were measured in 8 patients with acute liver failure and 20 healthy controls. Meanwhile to observe liver function and varicose vein of esophagus.

RESULTSIn cirrhotics, PBV and PBF increased immediately after transplantation [from (13.7 +/- 4.2) cm/s to (58.4 +/- 25.2) cm/s and from (958 +/- 445) ml/min to (3024 +/- 1207) ml/min respectively, P < 0.05]. HA-RI also augmented [from (0.65 +/- 0.11) to (0.74 +/- 0.12), P < 0.05]. PBV returned to normal values after 6 months, PBF returned to normal value after 2 years. Spleen size decreased significantly, but splenomegaly persisted after 3 years. In addition the esophagogastric varix ameliorated significantly.

CONCLUSIONSAbnormal splanchnic hemodynamic changes for patients with portal hypertension still will long-term exist after OLT, but does not effect recovery of hypersplenism, esophagogastric varix and liver function.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Hemodynamics ; Hepatic Artery ; physiopathology ; Humans ; Hypertension, Portal ; pathology ; physiopathology ; surgery ; Intraoperative Period ; Liver ; physiopathology ; Liver Transplantation ; Male ; Middle Aged ; Portal Vein ; physiopathology ; Splanchnic Circulation ; physiology ; Spleen ; pathology

Objective To summarize the clinical experience in liver retransplantation. Methods From June 2002 to December 2005,a total of 185 cases of liver transplantation were performed in our hospital,including 8 cases of retransplantation.Those clinical data were analyzed retrospectively. Results The rate of liver retransplantation was 4.32%.The average MELD scores before primary transplantation and retransplantation were 15.6 and 23.9,respectively.The average interval between primary transplantation and retransplantation was 316 days(78~725 days).Causes of retransplantation included 3 cases of biliary complications,2 cases of chronic rejection,1 case of hepatic artery thrombosis,1 case of acute rejection and 1 case of recurrence of hepatitis B.The former 3 cases died of severe infection combined with multiple organ failure 101,16 and 28 days after retransplantation.The latter 5 cases recovered smoothly,and have survived 27,12,8,4 and 3 months up to now. Conclusion Liver retransplantation is an effective way to save the patient with hepatic allograft failure.Good knowledge of the indications of retransplantation,careful selection of the operation time,excellent surgical skills and meticulous postoperative management will contribute to the success of liver retransplantation.

OBJECTIVETo construct one lentiviral vector containing mouse SRY-related silencing group--box gene 9 (SOX9) and to transfect murine bone mesenehymal stem cells (mBMSCs) in vitro and observe the expression of target gene.

METHODSRNA inteference target sequence was designed in connectin with mice SOX9 gene sequence. The double strands DNAoligo containing interference sequence were synthesized and cloned into lentivirus vector. The siRNA lentiviral vector with SOX9 gene silencing was constructed and identified, which was transfected into rat bone mesenehymal stem cells. The expression of target gene was detected by immunofluorescence, RT-PCR and Western blot.

RESULTSLenti-SOX9-siRNA-EGFP was recombined successfully and transduced efficiently into mBMSCs. The expression of SOX9 gene silencing was confirmed by RT-PCR and Western blot.

CONCLUSIONMouse SOX9 gene silencing by RNA interference and Lentiviral vector can transfected successfully into mBMSCs. Meanwhile,SOX9 gene may be silenced in SOX9 transduced mBMSCs. This will provide target cells for the following study about SOX9 gene respairing cartilage injury.

Animals ; Female ; Gene Expression ; Gene Silencing ; Genetic Therapy ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Mesenchymal Stromal Cells ; metabolism ; Mice ; SOX9 Transcription Factor ; genetics ; Transduction, Genetic