Objective To develop the human colon carcinoma(HT-29) liver metastasis model in nude mice,and to detect the expression of human telomerase reverse transcriptases(hTERT) mRNA in liver of nude mice. Methods Liver metastases were established in 30 Balb/c nude mice by intrasplenic injection of colonic cancer cells(HT-29),and the spleens were resected.After being sacrificed,the tumor growth was observed,and the pathological examinations were performed.The expression of hTERT mRNA in the livers of nude mice was detected by RT-PCR technique. ResultsHuman colon carcinoma(HT-29) liver metastasis model was developed in all the 30 nude mice,with the liver metastasis rate of 100%.The pathological results revealed the occurrence of liver metastases,and the expression of hTERT mRNA was positive in the liver tissues. Conclusion Intrasplenic injection of HT-29 cell is a reliable way for producing colonic cancer liver metastasis model,and the positive expression of hTERT mRNA in liver tissues indicates the significance of hTERT in the early diagnosis and treatment of colon carcinoma liver metastasis.

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 explore the causes and treatment of repeated vulvovaginitis in girlhood in order to improve its prevention and treatment.Methods Fifty-one girls with repeated vulvovaginitis(age≤10 years)admitted to The Second Affiliated Hospital of Sun Yat-sen University from Jan.1990 to Nov.2004 were reviewed retrospectively.Results We found 28 girls(55%)suffering from non-specific vulvovaginitis and 14 ones(27%)suffering from posterior recto-vaginal fistula with in 51 patients.Five girls(10%)were smitten with vulval ulcer and 3 ones(6%)had been were found with vaginal foreign bodies.One girl(2%) was smitten with adhesion of labia minora.The vaginal discharges taken from 21 girls were cultured. Seventeen cases found bacteria.The positive rate of bacteria culture in the 21 cases reached 81%,in which, E.coli accounted for 5 cases(24%),staphylococcus and streptococcus accounted for 3 cases(14%) respectively.Patients suffered from non-specific vulvovaginitis and vulval ulcer accepted external lotion, antibiotic ointment or combining with antibiotics.Patients suffered from posterior recto-vaginal fistula accepted fistulectomy.Three girls who found vaginal foreign bodies took out of foreign bodies by hysteroscopo.Fifty-one girls all were cured after appropriate therapy.Conclusions Vulvovaginitis is the most common gynecologic diagnosis in girlhood.The principal cause of repeated invasion is non-specific vulvovaginitis and the secondly one is posterior recto-vaginal fistula.It need overhaul during the diagnosis.It is very availability to use hysteroscopy and do bacteria culture+antibiotic sensitivity test for repeated pediatric vulvovaginitis.

OBJECTIVETo observe the effect of cyclic equiaxial tensile strain in the early differentiation of bone marrow mesenchymal stem cells(BMSCs) into cartilage in mouse under conditions of two-dimensional culture, and to investigate the mechanism of cyclic equiaxial tensile strain in early chondrogenic differentiation.

METHODSSixteen KM mouse aged 4 weeks were selected, male and female unlimited, with an average weight of 19.5 g (17 to 21 g). After extracting and isolating the BMSCs from KM mouse, then subculture the BMSCs to the 3rd generation. Seed the cells in the biological plate(BioFlex). According to experimental design, the cells were divided into 6 groups, blank group: ordinary culture medium was cultured for 8 days without isometric cyclic tensile strain stimulation. Control group: chondrogenic differentiation medium was used to culture for 8 days without isometric cyclic tensile strain stimulation. Experimental group: the experimental group was divided into 4 groups, all of which were cultured with chondrogenic differentiation medium for 8 days. During which isometric cyclic tensile strain stimulation was given for 1, 3, 5 and 7 days respectively. At the 8th day, all the cells were collected, the expression of the Sox9, Col-II and ROCK 1 signaling pathway-related molecules was analyzed by RT-PCR. Cells in each group were extracted, and the efficiency of cell proliferation in each group was detected by CCK-8. Glycosaminoglycan content in medium changed last was detected using ELISA. Pericellular matrix was observed by Safranin O staining and Alcian Blue staining. Normal measurement data using mean±standard deviation compared between the blank group and control group using paired t-test, compared between the experimental group and relative group using single factor analysis of variance.

RESULTS(1)After 8 days of culture, compared with the control group, the relative expression of Sox 9 and Col-II mRNA in the experimental group increased gradually with the increase of loading time(<0.05), while the relative expression of ROCK1 mRNA decreased(<0.05). Compared with the blank group, the relative expression of ROCK1 mRNA in experimental group and control group increased (<0.05). (2)With the increase of loading time, the experimental group showed a trend of decreasing at first and then increasing, but compared with the blank group and the control group, the control group decreased significantly. (3)Glycosaminoglycan content in the medium changed last was detected by ELISA. The glycosaminoglycans in the experimental group increased gradually, and the content changes on 7 days loading group were statistically significant compared with other groups(<0.05). (4)Safranin O and Alcilan staining showed that there was a tendency of cartilage differentiation in the experimental group, and the shape gradually increased with time, which was more obvious than that in the control group; The PCM, Col-II and GAG in the experimental group increased gradually with the increase of mechanical stimulation days, which were more obvious than those in the control group.

CONCLUSIONSUnder conditions of two-dimensional culture, in the early differentiation of mesenchymal stem cells into cartilage, cyclic equiaxial tensile strain can promote the proliferation of BMSCs and the differentiation into chondrocytes. Moreover, cyclic equiaxial tensile strain may promote chondrogenic differentiation through inhibiting the Rho/ROCK 1 signaling pathway.

Objective To explore the effects of modified tubo-uterine implantations performed on women with proximal tubal occlusive infertility after femal sterilization with mucilago phenol.Methods Two hundred and eight infertile women who were admitted to the Second Affiliated Hospital of Sun Yat-sen University between 1986 and 2004 were included.They all accepted modified tubo-uterine implantation after occlusion of fallopian tubes with mucilago phenol.Results It was found that the occlusions were all located in the interstitial portion or isthmic portion of the fallopian tubes.Different degrees of pelvic adhesions were found in 65 cases.Fifty-seven cases were slightly adhesive,seven cases were of moderate degree and one case was severe.One hundred and ninety-nine cases were followed up after operations(95.7%).One hundred and ninety-three women accepted hydrotubation in the following month just after the operation and 185 women were found to be unobstructed(95.8%).One hundred and forty-three women became pregnant, the pregnant rate being 71.9%(143/199).One hundred and twenty-five women had term deliveries (87.4%),three women were in early pregnancy and two in midtrimester pregnancy.Eleven women had spontaneous abortion(7.7%).Two women had tubal pregnancy(1.0%).None of the 199 cases had any signs of endometriosis.Conelusions Modified tubo-uterine implantations are quite effective for proximal tubal occlusive infertility.It may be a favorable method for such kind of tubal occlusions.

Objective To evaluate hemodynamic changes in liver treated by transjugular intrahepatic portosystemic stent-shunt(TIPSS)with hepatic computed tomography(CT)perfusion,Doppler ultrasound and portal vein pressure measurement,as well as the correlation among these methods.Methods Hepatic CT perfusion was performed in 9 cirrhotic patients one week before TIPSS and 72 hours after TIPSS. Intraoperative portal vein pressure was measured before and after portosystemic shunt establish.The follow- up hepatic CT perfusion were carried out in 3 cases at 3 months and 6 months postoperatively.The hemodynamic surveillance by Doppler ultrasound were performed in 48 hours and 3 months after TIPSS for 9 cases,and in 6 months after TIPSS for 6 cases.Two cases underwent venography and portal vein pressure measurement in 6 months after TIPSS treatment.Results The mean of portal vein perfusion(PVP),total hepatic blood flow(THBF),hepatic perfusion index(HPI)and portal vein free pressure(PVFP)before TIPSSwere(0.92?0.18)ml?min~(-1)?ml~(-1),(1.28?0.17)ml?min~(-1)?ml~(-1),(28?8)%,and (23.92?0.86)mmHg,respectively.In 72 hours after TIPSS,the mean of PVP,THBF,HPI and PVFP were(0.21?0.15)ml?min~(-1)?ml~(-1),(0.74?0.18)ml?min~(-1)?ml~(-1),(74 +13)%,and (12.62?1.54)mm Hg,respectively.After treatment,the mean of PVP was(0.49?0.05)ml?min~(-1)? ml~(-1)at 3 months and(0.57?0.03)ml?min~(-1)?ml~(-1)at 6 months,respectively.There was negative correlation between PVP and PVFP before TIPSS(r=0.678,P0.05).Moreover,a signifieant correlation was found between the degree of portal vein pressure decrease and portal vein perfusion decrease(r=0.867,P

In order to clarify the chemical constituents of Si-Wu Decoction rapidly and holistically, we analyzed the ethanol extract of Si-Wu Decoction by UPLC/Q-TOF-MSE and UNIFI which based on traditional Chinese medicine database, the probable structures of 113 compounds were identified. The results show that this method can rapidly and effectively characterize the chemical compounds of Si-Wu Decoction and provide a new solution for identification of components from complex TCM extract.
Chromatography, High Pressure Liquid ; Databases, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Medicine, Chinese Traditional

A high resolution Micro-CT system for small animal imaging is introduced in this paper. Micro-focus X-ray tube with focal diameter of 100 microm and flat panel detector with imaging area of 13cm x 13cm are adopted in this system. The data acquired in rotation scanning is reconstructed with cone beam algorithm. The resolving power of the detector is measured to be 31 lp/cm at 10% of the MTF. The resolution of the Micro-CT system could achieve 185 m when the magnification factor is 1.94. Thighbone of a rabbit is used as sample imaging with the system. The trabecular bone could be imaged clearly. And the ability of small animal imaging of the system has been demonstrated.
Animals ; Equipment Design ; Rabbits ; X-Ray Microtomography ; instrumentation

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

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult