Objective To observe the bacteriostasis of herba houttuyniae liquid in bladder irrigation for treatment of rabbit bladder infection model, and to offer a new way for prevention and treatment of urinary tract infection.Methods Twenty male New Zealand long-eared white rabbits were divided into gentamicin control group and three different concentrations herba houttuyniae groups according to random number table method, each group 5 rabbits. The rabbit bladder infection model was replicated by injecting coli liquid 1 mL (105 cfu/mL) into the urinary bladder through a urethral catheter, and clipping the catheter for 1 hour. After model establishment for 3 days, in the three herba houttuyniae groups, 0.25, 0.50, 1.00 g/mL herba houttuyniae liquid 50 mL were injected into the bladder respectively, and in the gentamicin control group, 160 kU gentamicin was injected into the bladder. Before molding and after molding for 3, 5, 7, 9 days, ear blood in rabbit was taken, and white blood cell (WBC) count and neutrophil percentage (N%) were tested; at the same time, the middle segment morning urine was taken to detect bacteriuria and urine WBC positive rates; after the experiment, the animals were sacrificed, the urinary bladder tissues were taken, and the pathological changes in urinary bladder tissue were observed by a light microscope.Results Before molding, the bacteriuria in animals of each group was negative and the results of urine WBC, blood WBC and N% were within normal range. After molding for 3 days, bacteriuria and urine WBC positive rates reached 100%; with the prolongation of treatment after molding, the above two rates in each group were gradually decreased, and herba houttuyniae 0.50 g/mL and 1.00 g/mL groups appeared statistical significance on the 5th day (bacteriuria positive rate was 60% and 40%, urine WBC positive rate was 60% and 40% respectively, while the gentamicin control group and herba houttuyniae 0.25 g/mL group was still 100% positive, allP < 0.01); on the 9th day, the bacteriuria positive rate in both higher dosage herba houttuyniae groups was 0, in 0.25 g/mL group, it still had the positive rate 20%, showing herba houttuyniae 0.50 g/mL and 1.00 g/mL had stronger inhibitory effect than herba houttuyniae 0.25 g/mL. WBC and N% were increased in each group after molding, and on the 3rd day, WBC and N were significantly higher in each group than those before molding [WBC (×109/L): gentamicin control group 13.00±5.23 vs. 8.95±2.69, herba houttuyniae 0.25 g/mL group 14.52±10.28 vs. 8.73±3.16, herba houttuyniae 0.50 g/mL group 12.97±11.33 vs. 7.83±4.51, herba houttuyniae 1.00 g/mL group 13.03±9.17 vs. 6.45±3.85. N%: gentamicin control group 80.99±14.93 vs. 63.41±7.96, herba houttuyniae 0.25 g/mL group 89.62±9.41 vs. 70.01±9.13, herba houttuyniae 0.50 g/mL group 87.69±11.34 vs. 67.75±8.91, herba houttuyniae 1.00 g/mL group 85.67±12.81 vs. 65.12±4.63]. On the 7th day, WBC and N% fell, and on the 9th day, the WBC and N% were restored to normal range in herba houttuyniae 0.50 g/mL and 1.00 g/mL groups [WBC (×109/L): herba houttuyniae 0.50 g/mL group 8.56±5.38, herba houttuyniae 1.00 g/mL group 7.93±4.82. N%: herba houttuyniae 0.50 g/mL group 65.97±8.35, herba houttuyniae 1.00 g/mL group 65.57±9.12]. Under light microscopy, it was shown that the morphological appearances of tissue cells in various layers of bladder mucosa were normal in herba houttuyniae group, demonstrating that herba houttuyniae fluid in bladder did not cause allergic inflammatory reaction, etc.Conclusion The bacteriostatic action of herba houttuyniae liquid for bladder irrigation in treatment of rabbits with urinary tract infection is better than that of gentamicin.

Objective To construct and identify lentiviral vectors carrying the connexin43 (CX43) gene. Methods Plasmids containing the CX43 gene and lentiviral vectors were digested using EcoRI/XbaI restriction enzymes, and the target gene fragments were cloned into the lentival vectors to result in CX43 recombinant lentiviral vectors (pHBLV-CMVIE-IRES-ZsGreen-CX43). CX43 recombinant lentiviral vectors were identified by restriction enzyme digestion and electrophoresis, and sequencing was carried on only for correct vectors after identification. The successfully-constructed CX43 recombinant lentiviral vectors and packaging plasmids were mixed and contransfected into 293FT cell for packaging and producting virus. Then the virus was collecting, concentrated and titrated in 293FT cells. Finally, the expression of CX43 gene was assessed by real-time polymerase chain reaction(PCR). Results The restriction enzyme digestion and electrophoresis and sequencing results proved that CX43 recombinant lentiviral vectors were constructed correctly. Lentiviral concentrated virus suspension titer was 1× 108/ml. The up-regulation expression of CX43 was detected correctly by real-time PCR in 293FT cells. Conclusions Stable lentiviral vectors expressing CX43 gene is constructed successfully.

BACKGROUND:Our previous studies have shown that a soft substrate has a significant effect on morphology and cytoskeleton of rat bone marrow mesenchymal stem cel.
OBJECTIVE:To explore the effect of polyacrylamide gels as soft substrates with different elastic moduli on the chondrogenic differentiation of human synovial-derived mesenchymal stem cels.
METHODS:The synovium was harvested from patients with osteoarthritis under sterile conditions, and primary human synovial-derived mesenchymal stem cels were separated using limiting dilution assay. The flow cytometry and multi-directional differentiation experiments were used to identify the cel surface markers and function of the human synovial-derived mesenchymal stem cels, respectively. The polyacrylamide gels with the elastic modulus of 0.4, 6, 30 kPa, which were made using various amounts of acrylamide and bis-acrylamide, were used to culture human synovial-derived mesenchymal stem cels under induction with transforming growth factor-β1 for 7 and 14 days. RT-PCR was used to test the expression of chondrogenic genes, type II colagen gene and cartilage acidic protein 1. The 6-wel cel culture plates served as controls.
RESULTS AND CONCLUSION: The human synovial-derived mesenchymal stem cels showed different cel morphology in the different elastic modulus of polyacrylamide gels. The expression of type II colagen gene and cartilage acidic protein 1 were affected by the different elastic modulus of polyacrylamide gels and culture time, and there was an interaction between these two factors. At 7 days of induction, the expression of cartilage acidic protein 1 gene on 6 kPa polyacrylamide gels was the highest (F=44.350,P=0.000); meanwhile, the expression of type II colagen gene on 0.4 kPa polyacrylamide gels was the highest (F=6.384,P=0.005). These findings indicate that polyacrylamide gels with lower elastic modulus are superior to routine culture plates to promote the chondrogenic differentiation of human synovial-derived mesenchymal stem cels.

Objective To study the occurrence,diagnosis and treatment of uveal metastasis in patients with breast cancer.Methods In 2400 cases of breast cancer patients,2 cases of uveal metastasis admitted from Jan.2007 to Aug.2011 in our hospital were retrospectively analyzed to study the occurrence,diagnosis and treatment.Results Both the 2 cases had the symptoms of blurred vision and hypopsia.Fundus examination and funds fluorescein angiography showed that the 2 cases had unilateral choroidal metastasis with the discovery rate of 0.08％.Case 1 was found to have uveal metastasis before breast cancer was confirmed and case 2 was found to have uveal metastasis during the 4th chemotherapy cycle.No other distant metastasis was found in these 2 patients.The eye symptoms improved obviously after chemotherapy.Conclusions In order to get a good therapeutic effect for breast cancer patients with uveal metastasis,we should pay attention to patients' symptoms and signs.Eyes examination should be taken when patients have eye symptoms and signs.

Objective: To explore and evaluate the biomechanic relationships between different depth of pedicle screw penetration with the sagittal plane reconstruction in thoracolumbar fracture. Methods:Six fresh cadaveric specimens of lumbar spine from L_1 to L_3 were used to make the model of thoracolumbar fracture. The system of universal spine system( USS )pedicle screw was adopted with the 6 mm diameter of screw. Each of two Schanz screws was implanted into the pedicles of L_1 and L_3 A canulated screw was fixed into the former of vertebral body in L_1 and L_3, and the distance of the two canulated screws was taken as the normal height. The axial loads were given while the pedicle screws were implanted at the depth of D1, D2 and D3, and the distance of the two canulated screw was measured as well as the distance was reduced to the normal height by axial load. The index measured included of the depth of pedicle screw penetration, the height of fractured vertebral body and afterload. Results: Along with the increasing of afterload, the height of injured vertebral body was increased accordingly, but the extent was different at three depth of pedicle screw penetration (D1, D2, and D3). While the injured vertebral body was reduced totally (reduced distance 0.00 mm), there was (2 630±13) g of afterload needed in Dl depth, and (2 339±61) g and(2 221± 164) g of afterload in D2 and D3 depth respectively. There was significant difference in distance between D1, D2 and D3 (P< 0.01), however, no significant difference between D2 and D3 (P> 0.05). Conclusion: There is a relationship in the depth of pedicle screw penetration, the capacity of reduction and sagittal plane reconstruction. The depth of pedicle screw had a significant effect on the capacity of reduction for the injured vertebral body, which would be the best option in biomechanics when the pedicle screw was implanted more than 1/2 pedicle or all of it.

Objective To investigate the role of bone metabolic indices in the diagnosis and monitoring of therapeutic effects in osteoporosis, the serum tartrate-resistant acid phosphatase form 5b(TRACP5b) level and other parameters such as Dpd in urine of osteoporosis animal model (ovariotomy rats) in different timing point were measured and analyzed. Methods Eighty female SD rats were divided into five groups and had ovariotomy operation (one group was pseudo-operated). Drug treatment began at the third day after the operation. Six rats in each group were sacrificed after 4 weeks and 14 weeks respectively, the bone metabolic indices were measured and bone histomorphometric data were collected. Results Four weeks after operation, the Dpd/Cr and TRACP5b in ovariotomy group were significantly higher than those in the pseudo-operated group, and TRACP5b was still higher after 14 weeks. In the drug treatment groups, serum E2 level was not changed 4 weeks later, however, Dpd/Cr and TRACP5b had changed greatly. After 14 weeks of drug treatment, there was only a significant decrease of TRACP5b among the bone resorption parameters, but other indices had no change.Bone histomorphometric data showed that the bone volume was improved in each drug treatment group. Conclusions Serum TRACP5b level can reflect the activitiy of osteoclasts and the situation of osteoporosis either in the early stage (high turnover period) or in later stage (low turnover period) of osteoporosis. The drugs used in this experiment can inhibit the ostoclast activity and reduce the level of bone resorption parameters, especially the TRACP5b level. Bone histomorphometric results suggestes that the function and manners of different drugs vary in improving of bone volume.

Objective To study ADFR with statin programmed therapy of osteoporosis in ovariectomized rats.Methods Fifty female rats were randomly allocated into 2 groups:sham operation (S, n =10) and OVX ( n =40) group.After operation for one month,OVX were randomly allocated into 4 groups (each n =10):OVX,statins (T),bisphosphonates (B) and statins+bisphosphonates+calcium+vitamin D (ADFR).After feeding statins or bisphosphonates or ADFR for 100 days,all rats were sacrificed.The effects of T or B or ADFR on bone histomorphology or osteocalcin in sera or deoxypyridoxine in urea were studied.Results The data showed that osteocalcin and deoxypyridine in OVX group were significantly improved compared with S group ( P 0 05) ,in B group was decreased,and in ADFR group was increased compared with OVX group.The histomorphometric date showed that TOS,MOSW,STS/DTS,TBOS,TBSC and iMAR in OVX were significantly increased,and TBV,MLT and ? in OVX were decreased,compared with S group.TBV in B,T and ADFR groups was larger than that in OVX group.TOS,MOSW,TBOS and TBCS in B group were smaller than those in OVX group,? in B group was longer than that in OVX group,TBCS and ? in T group were increased compared with OVX group.Conclusion Statins promote bone turnover,increase osteoblast activity and osteoid production,and reduce the bone construction.Bisphosphonates inhibit bone absorption,while ADFR acelerate bone formation and reduce bone loss,suggesting that polytherapy is preferable to monotherapy.

Objective:To demonstrate the relationship between the Th 1/Th2/Th17/Treg paradigm and the bone loss induced by estrogen deficiency and looking for potential target for clinical treatment.Methods:30 BALB/c mice were divided randomly into the normal control group , the sham operation group , and the ovariectomy group.The serum estradiol ( E2 ) was assessed by ELISA.Bone mineral density (BMD) of thigh bone was measured with dual energy X ray absorptiometry.Meanwhile,the T-cell subsets (Th1:CD4+TNFα+, Th2: CD4+IL-4+, Th17: CD4+IL-17 A+, Treg: CD4+CD25+Foxp3+) in spleen lymphocytes were analyzed by flow cytometry.Results:Compared with the normal group and the sham operation group , both E2 and BMD in the ovariectomy group decreased significantly ( P<0.05 ).The percentage of Th 1 and Th17 subset increased while the percentage of Th 2 and Treg decreased significantly in ovariectomy mice compared with sham operation mice.Correlation analysis showed that BMD was positively related to E 2 level and the percentage of Th 2 and Treg subset;however ,BMD was negatively related to the percentage of Th 1 and Th17 subset ( P<0.05 ).Conclusion: Conclusion: T-cell paradigm was involved in the bone loss induced by estrogen deficiency.Modifying T-cell paradigm may become a potential target for reducing bone loss induced by estrogen deficiency .

To investigate the relationship of bcl-2, p53, proliferating cell nuclear antigen (PCNA) to cell proliferation, apoptosis and pathological parameters, the patterns of cell growth and turnover in renal cell carcinoma (RCC), formalin-fixed and paraffin-embedded tissue blocks from 34 patients with RCC were examined. Cell proliferation activity was detected by PCNA immunostaining and the proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy- nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells. Expressions of bcl-2 and p53 were assessed immunohistochemically. Our results showed that the PI ranged from 6.0% to 24.0% (median 12.3%) and the AI from 2.0% to 8.0% (median 5.4%) in RCC. The expression of the bcl-2 protein was demonstrated in 15 cases (44.1%); the expression of the p53 protein, however, was seen in only 3 case. bcl-2 positivity was not associated with PI or AI or any pathological parameters. There were close associations between PI and tumor grade and stage, and a significant relationship between AI and the tumor grade of RCC, Our study suggests that bcl-2 positivity was not associated with PI or AI or any pathological parameters. There are close associations between PI and AI and tumor grade and stage of RCC. Active cell proliferation may be accompanied by frequent apoptosis in RCC.
Adult ; Aged ; Apoptosis ; physiology ; Carcinoma, Renal Cell ; metabolism ; pathology ; Cell Division ; Female ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics