Objective To clone and compare RNase HIIa and RNase HIIb of Chlamydia pneumoniae AR39(CpRNaseHIIa and CpRNaseHIIb). Methods Genes of CpRNase HIIs were amplified with the designed primers.Then,the recombinant plasmids were constructed,and CpRNase HIIs were expressed and purified with pET expression system.The 5′-32P-labeled RNA/DNA substrate and DNA with oligoribonucleotides substrates were prepared to identify characterization of CpRNase HIIs. Results Ribonuclease H activity of both CpRNase HIIa and CpRNase HIIb could cleave oligoribonucleotides strand,generating break with 3′-OH and 5′-phosphate ends.The results showed that there was different biochemical characterization between them. Conclusion CpRNase HIIa and CpRNase HIIb have different functions in C.pneumoniae.

Objective To investigate the epidemiological characteristics of the group A rotavirus (RV) among the children with diarrhea in Shenzhen ,in order to provide reference for clinical diagnosis and treatment .Methods Stool samples were collected for RV detection from children with diarrhea .The results were analyzed .Results A total of 3 509 cases of children with RV infection were checked out from 14 511 cases of children with diarrhea ,with the infection rate accounting for 24 .18% .RV infection occurred all year around ,and the infection peak was November ,December and January .RV infected children mainly distributed in the age group of 6 months to 2 years .Conclusion RV infection in children is a universal problem in Shenzhen .Clinic should pay attention to it .

High-fat diet-induced insulin resistance rat model was assessed by euglycemic-hyperinsulinemic clamp technique.Compared with the control group,the vaspin mRNA expression in adipose tissue was significantly decreased in insulin resistant rats induced by high-fat diet( P＜0.05 ),which was increased by metformin( P＜0.05 ).These data indicate that metformin may ameliorate insulin resistance in rats via upregulating vaspin mRNA expression.

Objective To investigate the effects of alcohol on hippocampal apoptosis-inducing factor (AIF) and caspase-3 expression in cercbral ischemia/reperfusion in rats.Methods Sixty-eight healthy adult male Wistar rats were randomly divided into a sham operation group (n =4) and a saline control group as well as low-dose (1.0 g/kg),medium-dose (1.5 g/kg) and high-dose (2.0 g/kg) ethanol groups.The saline control group and each dose alcohol group were redivided into ischemia/reperfusion 0,1,2 and 3 h subgroups according to the intervention time points (n =4 in each group).A model of middle cerebral artery ischemia/reperfusion in rats was induced by the suture method.The corresponding doses of ethanol or an equal volume of saline were injected intraperitoneally at ischemia for 2 h and reperfusion for 0,1,2 and 3 h in the alcohol groups and the saline control group.At ischemia for 2 hand reperfusion for 24 h,the neurological deficit in rats was evaluated by using behavioral score.Immunohistochemistry assay was used to detect the expressions of AIF and caspase3 in the hippocampus of ischemic sides at ischemia for 2 hand reperfusion for 24 h.Results The behavior evaluation showed that the neurological deficit score was 0 in the sham operation group.The neurological deficit scores in the different dose ethanol groups were significantly lower than those in the saline control group at the same intervention time points (all P=0.000),and there were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000).The high-dose ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P＞0.05).Immunohistochemistry revealed that the numbers of positive cells of AIF and caspase-3 in the sham operation group were 17.21 ±2.86 and 20.60 ±4.39,respectively,and they were significantly less than those in the saline control group and the each dose ethanol group at ischemia/reperfusion 0 h (all P =0.000); the number of positive cells of hippocampal AIF and caspase-3 in the different dose ethanol groups were all significantly less than those in the saline control group at the same intervention time points (all P =0.000).There were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000),and the high-does ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P =0.000).Conclusions Alcohol has protective effect on the cerebral tissue in ischemia/reperfusion injury in rats.Its mechanism may be associated with the inhibition of AIF and caspase-3 expression.

OBJECTIVE To seek for an effective,rapid,economic,safety and practical cleaning method,so as to ensure the high quality of sterilization in hospital supply center.METHODS Six hundred moderately contaminated recyclable instruments were divided into three groups randomly,conventional group,experimental group A and experimental group B(two hundred sorts each).The instruments in experimental group A were put into warmed water(40 ℃) for ten minutes first,and that in experimental group B were put into multi-enzyme solution for five minutes first,and then all the instruments in three groups were put into automatic sprinkling cleaning machine to accomplish the cleaning process automatically.After cleaning process,cleaning degree of the instruments and remained blood residues were compared in three groups.RESULTS The cleaning quality in the groups A and B was significantly higher than that of the conventional group,both P

AIMThe effects of angiotensin II on the changes of Ca2+ signal in cultured rat neonatal myocytes were investigated in order to reveal the localization and distribution of elementary Ca2+ signaling units.

METHODSThe cultured neonate rat myocytes were treated with angiotensin II, and calcium signal was detected using confocal laser scanning microscopy and fluo-4/AM calcium probe.

RESULTSThe propagation of Ca2+ waves was observed in rest and angiotensin II stimulated cardiac myocytes. Calcium fluorescent intensity oscillated slightly in myocytes and the average intensity was much higher in the nucleus than in the cytosol, all of which could be magnified significantly by AngII (10(-6) mol/L). Ca2+ oscillation induced by Ang II was completely blocked by NO donor sodium nitroprusside. AngII evoked Ca2+ sparks close to the cell surface membrane, and couldn't be abolished by sodium nitroprusside.

CONCLUSIONThere are spatiotemporal dynamics of Ca2+ signaling patterns such as Ca2+ wave, Ca2+ spikes, Ca2+ oscillation and the whole cell Ca2+ transients induced by angiotensin II, which might play very important roles in cellular cardiac function.

Angiotensin II ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium Signaling ; Cells, Cultured ; Microscopy, Confocal ; Myocytes, Cardiac ; drug effects ; metabolism ; Rats

OBJECTIVETo study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis.

METHODSFrom January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis.

RESULTSThe patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening.

CONCLUSIONPerfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.

Adult ; Angiography, Digital Subtraction ; Arthroplasty, Replacement, Hip ; Female ; Femur Head Necrosis ; therapy ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged

Fed with high-fat diet and assessed by hyperinsulinemia-euglycemia clamp technique, rat models with insulin resistance were successfully induced. Compared with normal chow group ( NC ), serum concentrations of free-fatty acids(FFAs) and baseline insulin in high-fat diet group(HF) was higher( P＜0.05 ), the average glucose infusion rate from 60 to 120 min( GIR60-120 ) was lower( P＜0.01 ), and the expression of p-IRE-lα in the liver was higher( P＜0.05 ). Furthermore, the expression of p-IRE-1α in the liver was positvely correlated with the serum concentration of FFAs. All these data indicate that high-fat diet may induce endoplasmic reticulum stress in the liver by elevating serum concentration of FFAs, and may participate in the genesis of insulin resistance via p-IRE-1α.

Objective To investigate the hepatoprotective effect of silybin-phosphatidylcholine complex(SPC) in sepsis rats.Me-(thods) Fifty Wistar immature rats were randomly divided into 3 groups:normal control(10 cases),septic group(30 cases) and interfe-(rence) group(10 cases).In septic group and interference group,rats were treated by cecal ligation and puncture(CLP).Meanwhile,the interference group was given SPC before 2 hours of CLP and after 2 hours of CLP.Blood of rats was taken from all groups to determine the levels of serum tumor necrosis factor-?(TNF-?),interleukin-1(IL-1),alanine aminotransferase(ALT) and aspartate aminotransferase(AST) at varying intervals.Results The levels of serum TNF-?,IL-1,ALT and AST after CLP were significantly elevated in septic group compared with the normal control group(P

ObjectiveTo explore the effect of estrogen on osteoblast apoptosis induced by serum hungry in vitro.MethodsOsteoblasts of second or third generation from newly born SD rats calvaria were divided randomly into the control group, serum hungry group and serum hungry with estrogen group. Cells of each group were incubated for 24 h, 48 h, 72 h, 5 d, 7 d and 14 d, then labeled using TUNEL staining and examined for morphological characteristics of apoptotic cell under light microscopy after incubated for 72 h. The rates of apoptotic cells of each group were examined with flow cytometry.ResultsThe cells of the control group showed normal appears, the serum hungry group had many cells with purple and blue particles in nuclei, but serum hungry with estrogen group had less such cells. The rate of apoptotic cell significantly increased in serum hungry group and decreased in serum hungry with estrogen group compared with the control group examined with flow cytometry (P<0.05).ConclusionEstrogen can repress osteoblasts apoptosis of rats induced by serum hungry.