Animals ; Humans ; Hypertension, Pulmonary ; complications ; Hypothermia ; complications ; Hypoxia ; etiology ; therapy ; Respiratory Distress Syndrome, Adult ; complications ; Shock, Septic ; complications

Objective To explore the effects of hypericin on inhibition of hepatitis 15 virus (HBV) replication.Methods The concentration gradient hypericin was added to HepG2.2.15 cell culture system and lamivudine was used as control.Enzyme-linked immunosorbent assay (ELISA) and Southern blot were used to examine HBsAg,HBeAg and HBV DNA level in the culture supernatant,respectively.Half inhibitory concentration (IC50) and half effective concentration (EC50) of hypericin were calculated.The effects of hypericin on HBV DNA polymerase were detected by 32p marked deoxy-ribonucleoside triphosphate as the substrate. Independent sample t-test and single factor analysis of variance were used to compare the data between two groups and among multiple groups.Results The inhibition rates of hypericin on HBsAg, HBeAg were enhanced with hypericin concentration increasing and those were higher than lamivudine control group when the concentration was higher than 0.5 μmol/L (t=-0.127,P＜0.05).Southern blot confirmed that hypericin was stronger in inhibition of HBV DNA (EC50=0.2 μmol/L) than lamivudine (t=-0.058,P＜0.05).Hypericin was non-toxic on HepG2.2.15 cells in the range of test with EC50 of 0.2 μmol/L and IC50 of 200 μmol/L.Hypericin did not act on HBV DNA polymerase which was quite different from lamivudine.Conclusions Hypericin can effectively inhibit HBV replication as well as antigen synthesis and is non-toxic on HepG2.2.15 cell.The anti-HBV target of hypericin is different from nucleos(t)ide analogues.

Objective To investigate the distribution of four common chronic diseases including essential hypertension, type 2 diabetes, coronary heart disease (CHD) and stroke, and their related risk factors in fertile and menopausal women. Methods Census data were collected by questionnaire, physical examination and laboratory examination in females older than 40 yrs in rural areas of Fangshan District, Beijing. Student's t test, chi-square test and logistic regression model were used. Results The prevalence rates of essential hypertension, type 2 diabetes, CHD, stroke in menopausal women (64.2%, 13.2%, 5.9%, 48.1%) were higher than those in fertile women (41.4%, 6.9%, 1.7%, 15.1%)with statistically significant differences (P

Objective:To explore the associations of menopause with cardiovascular disease (CVD) and related metabolic disorders (including hypertension,diabetes or higher blood sugar,obesity,dyslipi-demia)in Chinese females aged 45 -59 years .Methods:Data were acquired from a national cross-sectional survey conducted in 201 3,China,which was also the second follow-up survey of China Health and Retirement Longitudinal Study (CHARLS).In the study,4 702 Chinese perimenopausal women aged 45 -59 years were enrolled,including 1 769 premenopausal women and 2 933 postmenopausal women.Information was collected from questionnaires of health status and functioning and physical exa-mination.General liner models were employed to calculate age-adjusted or age-and-body-mass-index-adjusted or multiple-factor-adjusted means and their 95% confidence intervals (95% CIs)of cardiovas-cular risk factors (CRFs).The comparisons of CVD and its risk factors according to menopausal status, and calculation of adjusted odds ratios (ORs)and their 95%CIs for the associations of menopause with CVD and its risk factors were performed by multivariate Logistic regression models separately.Results:After adjustment for age and other confounders (including body mass index,marriage,education,current smoking,drink alcohol more than once per month),statistically significant associations of menopause with cardiovascular disease,which referred to having a history of heart disease or stroke in this study, were observed in the participants (OR =1 .34,95% CI:1 .04 -1 .74);prevalence of hypertension (OR =1 .42,95%CI:1 .1 0 -1 .84),prevalence of CRFs clustering number≥2 (OR =1 .31 ,95%CI:1 .02 -1 .68)and average waist circumference level (87.1 1 cm,95%CI:86.81 -87.42 cm in post-menopausal group vs.86.41 cm,95%CI:85.99 -86.84 cm in premenopausal group)were presented higher in postmenopausal group,compared with the premenopausal one.However,diabetes or higher blood sugar (OR =0.96,95%CI:0.60 -1 .52),dyslipidemia (OR =0.84,95%CI:0.59 -1 .20) and obesity (OR =1 .06,95%CI:0.86 -1 .32)were not shown significantly statistically related to me-nopause,after excluding effect of age and other confounders.Conclusion:Postmenopausal women in China had worse CRFs profile than the premenopausal ones,which implied menopause might aggravate the CRFs epidemic and increase the risk of cardiovascular disease beyond effects of aging,which would increase the CVD burden during and after their middle ages.

[Objective]To investigate the effect of selectively segmental cervical decompression through anterior approach in the treatment of senile segmental cervical spondylotic myelopathy. [Methods]Twenty-seven cases of senile segmental cervical spondylotic myelopathy experienced selectively segmental cervical decompression through anterior approach from 2004 to 2008 were reviewed retrospectively. Pre-and post-operative JOA scores were compared to evaluate the improvement of neurological function.[Results]Patients were followed up for an average of 19.7 months (ranged,18~27 months). No loosening or displacement was found in all cases. Postoperatively,ratio of JOA improvement was 56.1%,including excellent result in 8 cases (29.5%),good result in 11 cases (40.7%),fair result in 6 cases (22.2%),poor result in 2 cases (7.4%).[Conclusion]Selectively segmental cervical decompression through anterior approach is effective in the treatment of senile segmental cervical spondylotic myelopathy,giving consideration to patient's general physical condition and recovery of neurological function. Correct segments selection of preoperative decompression is the key point of result of postoperative restoration.

[Objective]To investigate the therapy of human enbryonic olfactory ensheathing cells(hOECs) transplantation on adult rats' transected spinal cord.[Method]Separated,cultured hOECS from abortus were obtained and percent age of P75 positive cells was calculated.Twenty-four Wistar rats were choosen and divided into experimental group and control group randomly.All rats of two goups were transected at the T_(10) segment of spinal cord.Nine to ten days later 5 ul(2.5?10~5) hOECs(labled by Hoechst 33342) were injected in the contusion area of lesion site in transplantation group.In control group same volume of DMEM-F12 were transplanted.1,2,4,6,8 and 10 weeks later all rats were estimated by system of the BBB locomotion score respectively.Then part of them were killed.The tissue section of spinal cords were separated and inspected by fluorescence microscope or the methods of immunohistochemical staining.Data were processed with statistical tools in double-blinded condition.[Result]Transplanted hOECs can survive for at least ten weeks and even migrated from the injuried area in spinal cord.From 4 weeks to 10 weeks the BBB locomotion scores of experimental group were improved significantly contrasting to that of control group(P

To estimate the protective effect of removing circulatory TNF-a immunoadsorpnon or Amberlite XAD-7 absorption respectively on endotoxin shock. Methods New Zealand white rabbits injected with lethal dose of endotoxin underwent hemoper-fusion through immunoadsorbent or Amberh'te XAD-7 respectively. Plasma TNF-a levels were detected, and the sorbent efficiency and survival rate were observed. Results After 2 hours hemoperfusion through immunoadsorbent or Amberlite XAD-7 respectively, plasma TNF levels were significandy lower than those in die control group, especially, die sorbent efficiency of immunoadsorp-tion was higher dian dial of Amberlite XAD-7 adsorption. Conclusion Compared widi die nonspecific Amberlite XAD-7 adsorption, immunoadsorption might be a specific and more effective therapy of endotoxin shock.

[Objective]To study the mechanism of tubular epithelial hypertrophy induced by high glucose.[Methods]Human tubular epithelium line HK-2 cells were cultured in DMEM/F12 medium containing 1 g/L glucose(normal control group),4.5 g/L glacose(high slucose group),and 1 g/L glucose+3.5 g/L mannitol(iso-osmia control group),respectively.The cells of every group after cultured for 24 h,48 h.and 96 h were collected.The mRNA levels of CTGF and p27~(kip) were detected by real-time PCR.The protein levels of CTGF and p27~(kip) were detected by Western blot.The cellular proliferative activities were observed by MTT.The total cellular protein contents were detected by Bradford method.The cell cycle process was analyzed by flow cytometry.The cells of every group after being cultured for 30 win,1 h,24 h,and 48 h were collected.The levels of phospho-ERK1/2 were detected by Western blot.Every step was repeated for 3 times.[Results]High slucose could significantly up-regulate the mRNA and protein levels of CTGF and p27~(kip),activate ERK1/2 signal conductive pathway in HK-2 cells,make more cells arrest in G1 phase,decrease cellular proliferative activities,increase total cellular protein content reflecting cellular hypertrophy.[Conclusion]CTGF is an important mediator of tubular epithelial hypertrophy induced by high slucose.The mechanism is that CTGF up-regulates the p27~(kip) expression through activating ERK1/2 pathway.Up-regulated p27~(kip) lead proliferative cell to be arrested in G1 phase and cause cellular hypertrophy.

Objective To observe the clinical efficacy of staphylococcal protein A immunoadsorption plus nonmyeloablative chemotherapy with CD34+ autologous peripheral blood stem cell transplantation in the treatment of refractory systemic lupus erythematosus (SLE). Methods Three patients with active SLE were enrolled into this study. All patients were diagnosed with lupus nephritis by renal biopsy and poorly responded to routine therapy. Before transplantation, patients were given 6 sessions of immunoadsorption apheresis using columns of staphylococcal protein A-silica with an interval of 3 days; each session processed 3 L plasma and a total of 18 L plasma was processed over the 6 treatments. Three days following the immunoadsorption apheresis, the mobilization of stem cells was realized by intravenous cyclophosphamide at a dose of 2 g per square meter of body surface area and subcutaneous recombinant human granulocyte colony-stimulating factor (G-CSF) at a dose of 5 g per kilogram of body weight per day for 5 days. Then, peripheral blood raonoclonal cells were obtained by CS-3000 Cell Separator, and passed through the Clini Macs CD34+ cell selection device, with the final concentration of CD34+ cells being 2.6×106, 2.1×106 and 2.4×106 per kilogram of body weight respectively, and that of CD3+ cells being 3×105, 2.1×105, and 2.0×105 per kilogram of body weight, respectively, in these three patients. The conditioning regimen consisted of oral fludarabine of 50 mg/d for 5 days plus intravenous pig anti-human thymocyte immunoglobulin (ATG) at a daily dose of 90 mg/kg for 5 days. After 72-hour treatment with ATG, the frozen stem cells were infused back to the patients. Clinical manifestations and lupus-correlated immune parameters were compared in patients at baseline and after transplantation. Results Following immunoadsorption apheresis, an obvious decrease was observed in the level of serum anti-dsDNA, antinuclear antibody and IgG antibodies, while an increase in the level of serum complement 3. All patients achieved the reconstruction of hemopoiesis 2-3 days after the transplantation. Also, an apparent clinical remission was achieved with the SLEDAI score being less than 3. Six months after the transplantation, serum anti-dsDNA and antinuclear antibodies as well as urine protein were undetectable, the level of complement 3 reached the normal range, and renal function was restored. Conclusions Staphylococcal protein A immunoadsorption plus nonmyeloablative CD34+ autologous peripheral blood stem cell transplantation are effective and safe for refractory SLE, but the long-term effect remains to be connfirmed by further studies.

BACKGROUND: Previous studies demonstrated that recombinant human bone morphogenetic pretein-7 (hBMP-7) can effectively promote the synthesis of extracellular matrix, repair of damaged disc matrix, restore of degenerative disc height. It is hoped that BMP-7 can be used to control and reverse the intervertebral disc degeneration. However, because of the short half-life of recombinant protein and low biological activity, it is difficult to maintain BMP-7 high concentrations on degenerative disc. Gene therapy can prevent these defects effectively.OBJECTIVE: To study the effect of hBMP-7 gene transfeotion on biological activity of primary cultured rabbit nucleus pulposus cells in vitro, to determine the feasibility of hBMP-7 gene which will on gene therapy research of intervertebral disc degeneration,and to provide basis for further study.DESIGN, TIME AND SETTING: A randomized and controlled observation was performed at the Institute of Cardiothoracic Surgery of Changhai Hospital from December 2005 to September 2006.MATERIALS: Six healthy New Zealand white rabbits of either gender, averaging 4 weeks old and weighing 500 g, were used in this study, and Ad-hBMP7 was constructed by the Cardiothoracic Surgery Institute of Changhai Hospital.METHODS: After rabbits sacrifice under aseptic condition, the nucleus pulposus was got. After digested with Pronase, type Ⅱcollagenase and type Ⅱ DNAase (4 hours, 37 "(2), the cells were seeded in the Petri dishes and were kept in the incubator. After 7 days and then twice a week, the media were refreshed. The nucleus pulposus cells were infected by adenovirus integrated with hBMP7 gene. The cells which were transfected by adenovirus vector for Lac-Z gene and which were not transfected were adopted as control.MAIN OUTCOME MEASURES: The expression of hBMP-7 was determined by RTopCR and Western blot. The effect of hBMP-7 on cell proliferation was surveyed by MTT. Furthermore, the effect of hBMP-7 gene on synthesis of proteoglycan and type Ⅱcollagen was detected by modified dimethylmethylene blue method and ELISA, respectively.RESULTS: Gene sequencing and PCR showed that hBMP-7 gene was inserted correctly and no mutation happened. The pdmary cultured rabbit nucleus pulposus cells were identical with those reported in literature. After Ad-hBMP7 transduct into the nucleus pulposus cells, high level of hBMP-7 expression was observed and lasted over 3 weeks. Also hBMP-7 gene can promote cell proliferation and synthesis of proteoglycan and type II collagen with significant difference compared with control groups (P < 0.05).CONCLUSION: hBMP-7 gene mediated by adenovirus can be the target gene for the further study on gene therapy of intervertebral disc degeneration.