Objective To study the activation effect of HBV RNase H protein on the transcription of cellular apoptosis susceptibility gene (CAS). Methods The promoter of DNA sequence of CAS gene was identified in GenBank by bioinformatics and amplified from HepG2 genome by PCR using sense (5′-GGTACCCGATTACATGTTGTACATGAAGG-3′) and antisence (5′-CTCGAGGCTGAGTTCCATTGCTATAG-3′) primers. As these primers contained Kpn I and Xho I recognition sites on their respective 5′-ends, the amplified DNA fragments were tested by sequencing and then subcloned into Kpn I/Xho I sites of pCAT3-Basic reporter vector by routine molecular biological methods. The reconstructed plasmid named pCAT3-CASp was identified by enzyme digestion of Kpn I/Xho I, in which the expression of chloramphenical acetyltransferase (CAT) was under the control of the promoter of CAS. The HepG2 cells were transfected by pCAT3-CASp, and then co-transfected by pCAT3-CASp and pcDNA3.1(-)-RH plasmids. At the same time, the empty pCAT3-basic and pCAT3-TXNRD1p were transfected (self-contructed by the authors) as controls. After 24h culturing, cells were collected and the expression of CAT activity was detected by ELISA according to the manufacturer′s protocol. Results The optical density of expression of CAT of pCAT3-CASp was 0.043 by ELISA, in contrast, the optical density of expression of pCAT3-Basic was 0.024. The expression of CAT in co-transfection of pCAT3-CASp and pcDNA3.1(-)-RH(0.065) was 1.5 times as higher as pCAT3-CASp plasmid (0.043), and 2.7 times as higher as pCAT3-Basic. Conclusions The CAS gene promoter identified in present study has transcription activity and HBV RNase H protein may activate the expression of CAS gene.

STUDY DESIGN: Changes of morphology and phenotype of cultured cells in media added lactate were observed. OBJECTIVES: To evaluate the effect of lactate on morphology and phenotype of cultured intervertebral disc cell. SUMMARY OF LITERATURE REVIEW: It was reported that lactate and pH were important factor in the degeneration of intervertebral disc. However the effect of lactate on morphology and phenotype of cultured intervertebral disc cell have not been studied. MATERIALS AND METHODS: Cells were dissociated enzymatically from rabbit nucleus pulposus. After attaining monolayer growth, the cells were incubated in media added 2mM or 5mM lactate. Total cell counts and morphological changes of the cells were periodically observed. Changes in cell phenotype were investigated by use of anti-collagen antibody stain. RESULTS: The cell groups added no lactate and 2mM lactate showed no difference in cell counts, morphology and phenotype. The cell group added 5mM lactate showed a reduction in final cell Counts and highel'ratio of fibroblastic cell in total population. Anti-collagen I Ab stained the Intra-and extra-cellular area of fibroblastic cells and intracellular area of chondrocytic cells. CONCLUSIONS: The current study suggests that high concentration of lactate inhibit intervertebral disc cell proliferation and accelerate morphological and phenotypical change to fibroblastic cell.
Cell Count ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; Hydrogen-Ion Concentration ; Intervertebral Disc* ; Lactic Acid* ; Phenotype*

Objective To investigate the effects of C-terminal fragment of parathyroid hormonerelated protein (PTHrP107-139) on bone mineral density (BMD), bone histomorphometry and biomechanical properties in ovariectomized (OVX) rats and its effect on bone metabolism is also explored. Methods Forty 4-month old female Wistar rats in which 30 were ovariectomized and then divided into 3 groups: the placebo, the PTHrPC and the CT groups, the other 10 rats were Sham-operated as the control group (Sham). Five weeks later, the rats of PTHrPC and CT groups were subcutaneously injected with PTHrP107-139 (40 μg/kg) and Salmon Calcitonin (15 U/kg) respectively once every other day. The rats of the placebo and sham groups were injected with 0.2 ml saline once every other day. After treatment of 12 weeks, all rats were sacrificed and all samples were collected and analyzed. Results ① Compared with the placebo, the BMD and bone strength of PTHrPC and CT groups were significantly increased (P<0.05). ② Histomorphometry revealed that the tetracycline labeled bone surfaces, osteoid surfaces, mineral apposition rate and bone resorption rate were remarkably decreased in PTHrPC, and CT groups comparing with those of the placebo group. Conclusion Cter-minal PTHrP107-139 is effective in increasing the BMD, bone strength and quality when administered intermittently to ovariectomy-induced osteoporotic rats. Its increasing in bone quality may relate to reducing bone turnover and inhibiting resorption.

Objective To investigate the therapeutic effects of human parathyroid hormone related protein (PTHrP1-34) on osteoporosis of ovariectomized osteoporotic rats. Methods Sixty 4-month-old female Wislar rats were involved in this study and 40 of them were ovariectomized and another 20 received sham operation. After 6 weeks of ovariectomy the osteoporosis model was confirmed by examing 10 ovariectomized and sham-operated rats. The 30 osteoporotic rats were randomly divided into 3 treatment groups, i.e. PTHrP, estradiol and placebo. Human 40 ?g/kg PTHrP1-34 was subcutaneously injected once daily to PTHrP group and the estradiol group was injected with 40 ?g/kg estradiol benzoate once every 3 days.The placebo and shamoperated rats were given 0.2 ml saline every 3 days. The bone mineral density (BMD), bone histomorphology, the bone weight of dry and ash and serum Ca,P,alkaline phosphatase (ALP) were measured after 3 months' therapy. Results After 6 weeks of ovariectomy, the lumbar BMD of ovariectomized rats were significantly declined compared with those of the sham-operated rats. After 12 weeks treatment the femoral and lumbar BMD and the rate of bone weight of dry and ash in the PTHrP group were increased obviously compared with those of placebo groups.There was no significant difference between PTHrP group and estradiol group, in PTHrP group the percent age of trabecular area,trabecular width,osteoblast surface and mineral apposition rate were obviously higher than those in placebo group.Conclusion Treatment with 40 ?g/kg dose of hPTHrP1-34 administered once daily is effective in treating ovariectomy-induced osteoporosis.

Background The effectiveness of medical tissue adhesive for the reparation of operative incision has been recognized,but its influence to ocular surface microenvironment is not quite clear.Objective This study was to appraise the safety and efficacy of α-cyanoacrylate,a medical issue adhesive,for the reparation of the conjunctival and skin laceration.Methods Twenty healthy New Zealand rabbits were randomly divided into suturing group and medical glue group.Routine ophthalmic examination,the level of tear secretion and breakup time of tear film (BUT) were tested before operation.The upper bulbar conjunctiva of each right eye was cut apart about 1.0 cm after surface anesthesia and the skin of back was incised about 2.5 cm length after local anesthesia.The conjunctival and skin lacerations were adhered with compound medical adhesive in the medical glue group,and continuous suture was done to repair the conjunctival laceration and interrupted suture to the skin laceration with 5-0 chorda serica chirurgicalis in the suture group.The routine ophthalmic examination,level of tear secretion,BUT and the states of wound healing were examined 1 day,2 days and 7 days postoperatively.Pathologic examination of the corresponding tissues was also carried out 1 week after the animals were sacrificed.The data were analyzed by SPSS 13.0 statistical software.This experiment followed the Administration of the Care and Use of Experimental Animals of Shanghai City.Results The conjunctival and skin lacerations healed well both in the suture group and the medical glue group.The levels of tear secretion 1 day and 2 days postoperatively in the medical glue group were (12.70±2.21)mm and (12.70±2.00)mm respectively,showing a significant lowness in comparison with (14.90±2.38)mm and (14.90±2.33)mm of the suture group (q =-4.02,P =0.03;q =-4.02,P =0.03).Compared with the baseline,the difference of the levels of tear secretion in the medical glue group was not statistically significant in various time points after operation (P=1.00,1.00,0.51).The BUT values at 2 days and 7 days postoperatively in the medical glue group were (4.50 ± 1.18) seconds and (4.10±0.88) seconds respectively,being significantly longer than (3.30 ± 1.06) seconds and (3.00±1.25) seconds in the suture group (q=4.37,P=0.02;q=4.19,P=0.03).Compared with the baseline,there was not statistically significant difference at each time point postoperatively in the medical glue group (P =0.28,0.59,0.21).However,BUT at each time point after operation in the suture group was significantly shorter than that of the baseline (P=0.01,0.01,0.00).Pathological examination showed that all the conjunctival and skin lacerations healed well.Trivial collagen hyperplasy was seen in the eyes of the suture group and infiltration of a few of inflammatory cells was found in the medical glue group.In the pathological result of the skin test,there was conspicuous scar and severe collagen hyperplasy in the suture group,but in the medical glue group,the skin structure was almost normal.Conclusions α-Cyanoacrylate is safe and effective for the repair of the conjunctival incision with little affection to ocular surface microenvironment.

OBJECTIVE: To analyze the general and forensic pathological characteristics of death due to fat embolism syndrome (FES) and to provide reference data for forensic identification. METHODS: Twenty autopsy cases due to FES were selected from the forensic center of a medical college from 1999 to 2012. The general and forensic pathological characteristics such as the ways and types of injuries, clinical manifestation and the pathological changes were summarized. RESULTS: Fat embolism mainly occurred after long bone fracture or a large area of soft tissue injury with the majority of cases being fat embolism of lung and occasional cases being combined embolisms of lung and brain as well. The onset of symptoms appeared shortly after the injury or surgery. Lipid droplets could be observed within small pulmonary vessels and verified by special staining. CONCLUSION There are particular characteristics in death due to FES in concern with types of injuries, onset of symptoms and pathological findings. In order to find out the direct evidence of FES, special staining (oil red O staining) can be used in the forensic identification.
Autopsy ; Cause of Death ; Death ; Embolism, Fat/mortality* ; Forensic Pathology ; Fractures, Bone/complications* ; Humans ; Male ; Middle Aged ; Pulmonary Embolism/etiology* ; Soft Tissue Injuries/complications*

No abstract available.
Humans ; Infant, Newborn* ; Intracranial Hemorrhages* ; Vitamin K* ; Vitamins*

BACKGROUND: The implementation of intraoperative normovolemic hemodilution is a strategy used in an attempt to diminish the need for or obviate allogeneic transfusion and to avert the potential complications. The goal of this study was to evaluate the safety and efficacy of moderate intraoperative normovolemic hemodilution. METHODS: Fifteen patients scheduled for posterolateral spinal fusion underwent intraoperative normovolemic hemodilution with 10% pentastarch to a target hematocrit level of 25% (hemodilution group). All units of blood procured by hemodilution and additional allogeneic blood was transfused in the perioperative period to maintain the hematocrit level of>25%. We investigated the effect of hemodilution on whole blood coagulation as measured by the thromboelastography and, evaluated its efficacy of decreasing the need for allogeneic blood transfusion as compared to the nonhemodilution group, retrospectively. RESULTS: Hemodilution with pentastarch caused a decrease in response and coagulation time (p<0.01) but did not influence on the alpha angle and maximum amplitude on thromboelstogram. Amount of transfusion of allogeneic blood was 4.1 1.7 units for the hemodilution group and 5.0 1.5 units for the nonhemodilution group. Net red blood cell volume ""saved"" from hemodilution was about 120 ml. CONCLUSION: Moderate intraoperative normovolemic hemodilution with pentastarch does not affect the coagulability of whole blood. But its efficacy of decreasing the need for allogeneic blood transfusion is minimal.
Blood Coagulation ; Blood Transfusion ; Erythrocytes ; Hematocrit ; Hemodilution* ; Humans ; Hydroxyethyl Starch Derivatives ; Perioperative Period ; Retrospective Studies ; Spinal Fusion* ; Thrombelastography

We reviewed 9 eyes which underwent combined phacoemulsification and trabeculectomy at separate site. Phacoemulsification was performed using corneal incision and trabeculectomy was performed at one superior quadrant using a limbus-based conjunctival flap with mitomycin C(0.2mg/ml, 3-5 minutes). Six eyes received 3-piece folable silicone lenses and 3 eyes received 1-picec PMMA(diameter 5.5mm) lenses. Mean preoperatie intraocular pressure (IOP) was 23.4+/-8.7mmHg. Mean follow-up period was 12.7(range 6-21)months. The mean postoperative IOP at 1, 6, 12 months were 14.6+/-4.8mmHg, 13.8+/-4.4mmHg, 16.6+/-6.8mmHg, respectively. Seven eyes achieved a final fisual acuity of 0.7 or better. One patient developed vision-impairing complications; flat anterior chambe and anterior dislocation of silicone lens. Other complications include posteror synechiae(4 eyes), capsular opacities (3 eyes), lens decentration with capsular contracture(1 eye). These rusults suggest that combined corneal incision phacoemulsification and trabeculectomy at separate site offers an effective technique to control postoperative intraocular pressure in patients with coexsisting cataract and glaucoma, though this procedure is not devoid of complications.
Cataract ; Dislocations ; Follow-Up Studies ; Glaucoma ; Humans ; Intraocular Pressure ; Mitomycin ; Phacoemulsification* ; Silicones ; Trabeculectomy*

We report a case of primary cutaneous cryptococcosis on the left knee of a 67 year-old woman. She had a large ulcerated and indurated plaque with yellowish purulent exudates on her left knee. A histopathological examination from the lesion showed numerous encapsulated, round spores. Cultures from the lesion showed the presence of Cryptococcus neoforrnans. This may have resulted from an immunosuppressive state due to long-term use of oral corticosteroids.
Adrenal Cortex Hormones ; Cryptococcosis* ; Cryptococcus ; Exudates and Transudates ; Female ; Humans ; Knee ; Spores ; Ulcer