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 effects of Xinshang Xuduan Decoction(XXD, mainly composed of Rhizoma Drynariae, Radix Dipsaci, and Eupolyphaga seu Steleophaga) on the proliferation and differentiation of rat osteoblasts and bone morphogenetic protein 2(BMP-2)expression in the osteoblasts . Methods The animal serum containing XXD was prepared by serum pharmacological method and then was mixed together with α-MEM for the cell culture. Osteoblasts were isolated from the skull bone of SD neonatal rats by collagenase digestion and were identified by alkaline phosphatase(ALP) staining and alizarin red staining. The third and fourth generations of osteoblasts were treated with XXD at the volume fraction of 5%, 10%, 20% for 24, 48 , 72 hours respectively, and then the proliferation of the cells was evaluated by Cell Counting Kit-8(CCK-8). In the other test, osteoblasts were cultured with blank control serum, and 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae, respectively, and then the ALP activity was examined by using ALP assay kit and the expression of BMP-2 was investigated by enzyme-linked immunosorbent assay(ELISA). Results XXD had a dose- and time-dependent effect on the proliferation of rat osteoblasts in a suitable volume fraction range from 5% to 20%, and the effect of XXD at 10% was the best. Compared with the blank control serum group, ALP activity was increased in the cells treated with 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae(P<0.01). On culturing day 7, the expression of BMP-2 was increased in 10% XXD group and Rhizoma Drynariae group(P<0.05). Conclusion XXD can increase the ALP activity and BMP-2 expression in the osteoblasts in vitro, so does the single herb of Rhizoma Drynariae. And their therapeutic mechanism in promoting the healing of fractures may be related with the enhancement of osteogenesis of osteoblasts.

This study discussed the dual affiliated institutions of one author in a scientific paper.This issue could give rise to troubles such as the dispute over signature rights, dispute over intellectual property rights, etc. Moreover, it could bring negative effects on the development of science. In addition, some suggestions were put forward to deal with this issue.

Objective To introduce our experience of performing posterior or anterior renal lip incision for partial nephrectomy in the treatment of renal hilum endophytic renal cell carcinoma.Methods From Jan.2010 to Jan.2014,five female patients with renal hilum tumors were treated in our institute.The median age was 54 (51-72) years.The median tumor diameter was 4.0 (2.8-4.8) cm.The median preoperative creatinine was 53.9 (52.6-75.4) μmol/L.One of them was solitary kidney with absolute indication; three cases had basic disease with relative indication; one was with selective indication.The patients were put in supine or lateral position.After general anesthesia,we preformed partial nephrectomy by cutting posterior renal lip in 3 cases and the anterior lip in 2 cases.We clamped the renal artery,opened the renal posterior or anterior lip,then dissected the tumor beside the pseudo-capsule.After removing the tumor,we used 3-0 absorption suture to control bleeding and repair the opened collecting system.Finally,we used 2-0 absorption suture to close the renal defect.Results The median operation time was 195 (155-215) min; the median renal warm ischemia time was 35 (15-70) min; the median postoperative hospital stay was 8 (7-9) d.There was no secondary bleeding and urine leakage happened.The pathological results showed that 3 cases with clear cell carcinoma,1 with papillary carcinoma and 1 with renal medullary interstitial cell tumor.All patients showed normal kidney shape.The median postoperative creatinine was 63.0 (59.4-75.4) μmol/L.After a median follow up of 24.2 mon,all patients survive without tumor recurrence.Conclusions The short-term result of posterior or anterior renal lip incision partial nephrectomy in treating endophytic renal hilum endophytic renal cell carcinoma is safe and feasible.

OBJECTIVETo investigate the application and effects of external fixator under the guidance of damage control therapy in limb open fracture.

METHODSFrom May 2008 to January 2013,72 cases with severe limb open fractures (Unincor- porated shock) were divided into control group and observation group (36 cases in each group). In observation group, 36 patients (including 22 males and 14 females) were treated by external fixator at stage I, as soon as possible after waiting for patients physiology conditions being stable,the stage II fracture operation was performed. In control group, 36 patients (24 males and 12 females) were treated by the first stage open reduction. The hospital admission time, open fracture severity score (OFSS),operation duration,operative blood loss,X-ray expose times, callus appear time, fracture healing time, postoperative infection rate, complications and Johner-Wruhs accceccment were recorded and evaluated.

RESULTSAll patients were followed up for 8 to 12 months (10.4 in averaged). The mean operation duration, operative blood loss, callus appear time, fracture healing time the mean operation duration reespectively were (56.79±8.87) min, (216.16±18.21) ml, (5.32±0.71) weeks, (12.79±2.52) weeks in observation group,and (104.53±9.28) min, (439.93±14.65) ml, (4.97±1.26) weeks, (14.81±2.63) weeks in control group. According to Johner-Wruhs acccecement,there were were 33 cases in excellent,2 in good,and 1 in poor in observation group,non-union of fracture in 1 case,local infection occurred in 1 cases; in control group 25 cases in excellent,6 in good, and 5 in poor, non-union of fracture in 1 case,local infection occurred in 8 cases. There were significant differences in operation duration, operative blood loss, callus appear time, fracture healing time, postoperative infection rate, complications (P<0.05). There were no significant differences in hospital admission time, open fracture severity score and X-ray expose times (P>0.05).

CONCLUSIONUnder the guidance of damage control therapy,using external fixator measures to treat patients with open fractures of limbs is worth popularizing in clinical application because it can shorten the operation time, less blood loss, reduce the infection rate and complications, improve the success rate of surgery and recovery rate.

Adult ; Aged ; External Fixators ; Extremities ; injuries ; surgery ; Female ; Fracture Healing ; Fractures, Open ; surgery ; Humans ; Male ; Middle Aged ; Operative Time ; Postoperative Complications ; epidemiology

Hip fracture patients preoperative and postoperative exist hidden blood loss which often affect patients' wound healing, increase the probability of infection, prolong rehabilitation exercise, influence postoperative effect. At the same time, the body's blood loss increase the activation of the blood clotting mechanism, promote the incidence of deep vein thrombosis, bleeding and deep vein thrombosis has become the main causes of high risk in hip operation. It is very important to stop bleeding, anticoagulation should not be ignored, so how to effectively deal with the prominent contradiction between the postoperative anticoagulation and bleeding or looking for a best balance has become a intractable problems in hip fracture treatment.
Anticoagulants ; therapeutic use ; Blood Loss, Surgical ; prevention & control ; Hip Fractures ; complications ; surgery ; Humans ; Intraoperative Complications ; drug therapy ; etiology ; prevention & control

Objective To establish KIF4A 3'UTR esiRNA library and analyze the advantage of method in studying the function of KIF4A in SGC-7901 cells.Methods The GST-fusion protein of Escherichia coli endoribonuclease Ⅲ (GST-RNase Ⅲ) was used to digest the double strand RNA (dsRNA),which was transcribed in vitro from a 502bp template of KIF4A genome (T7-KIF4A 3'UTR).The KIF4A esiRNA library generated from the above method and chemically synthesized KIF4A siRNA were then used to transfect SGC-7901 cells at 10 nmol/L and 20 nmol/L.Real-time quantitative PCR and Western Blot were used to detect the mRNA and protein level of KIF4A,respectively.Results The KIF4A esiRNA library was effectively established from dsRNA digestion using GST-RNase Ⅲ.KIF4A expression was significantly reduced in SGC-7901 cells transfected with KIF4A esiRNA or siRNA.In addition,the inhibitory effect of KIF4A esiRNA was more effective than that of chemically synthesized siRNA.Conclusion KIF4A esiRNA library which was obtained using biological method can more effectively inhibited the expression of KIF4A more effectively in SGC-7901 cells than chemically synthesized siRNA.Therefore,esiRNA library can be used as a new and more effective method in studying the function of KIF4A in SGC-7901 cells.

Objective To investigate the expression and significance of death receptor 4(DR4) and DR5 in human craniopharyngioma.Methods The expression of DR4 and DR5 was determined by immunohistochemistry and in situ hybridization in 28 samples of craniopharyngioma and 25 samples of normal brain tissue.Results With low expression in partial normal brain tissue,DR was expressed highly in all of the craniopharyngioma samples.High DR expression in craniopharyngioma tissue differed from low DR expression in normal brain tissue(P0.05).Conclusions High DR expression is prevalent in craniopharyngioma tissue.This may contribute to the apoptosis-induced therapy of craniopharyngioma.The control of DR expression lays in protein level.This may contribute to the selective induced-apoptosis of tumor necrosis factor-related apoptosis-induced ligand.

OBJECTIVE: To investigate the resistance phenotype of acinetobacter baumannii and the expression of ade efflux pump gene. METHODS: Non-repetitive 51 strains of Acinetobacter baumannii were collected in Peking Union Medical Hospital between Feb. 2004 and Feb. 2005. The active efflux system adeB structural gene and sequence were identified by PCR. RESULTS: The tested strains were not susceptible to common broad-spectrum antibiotics. Multidrug resistant Acinetobacter baumannii carried adeB active efflux pump gene. CONCLUSION: The multiple-drug resistance of Acinetobacter baumannii is independent of ?-lactamase. It maybe related to other drug resistance mechanism. The effect of active efflux system is possibly one of the major mechanisms of multidrug resistance of acinetobacter baumannii.

Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.