HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

Objective: To study the effects of the melt casting and the sintering methods on the machinability and other characteristics of K_2O-MgO-MgF_2-SiO_2 glass-ceramics. Methods: X-ray diffraction(XRD) and scanning electron microscopy (SEM)were used to examine the main crystal and the microstructure of the glass ceramics of the same composition which were made by melt casting and sintering methods, respectively. The transmissivity, flexural strength and brittleness index were observed respectively. Results: The glass ceramics made by two different methods have the same crystal, KMg_(2.5)Si_4O_(10)F_2, while the specimen prepared by the sintering method had lower transmissivity,better machinability and mechanical properties due to the higher mica volume percent. Conclusion; For production of fluoro-mica glass ceramics, sintering method has superiority compared to melt-casting method.

AIMStudy on the relationship between the degraded spinal cord injuries and the changes of the motor evoked potentials (MEP) to prove the diagnosis and prognosis value of MEP.

METHODSAfter injury at T8-T9 cord using modified Allen's weight-drop method, 27 male SD rats were divided randomly into control group (n = 5), group A (50 gcf, n = 8), group B (70 gcf, n = 8) and group C (100 gcf, n = 6). MEPs elicited by monopolar transcortical stimulation were recorded continuously before injury, just after injury, 15 minutes, 1 hour, 3 hours and 6 hours after injury. The rate of the size of the bleeding or necrosis area to the total cord was also calculated.

RESULTSMEP had no significant change in the control group. The amplitude of MEP's early components in group A or group B decreased or even obliterated after SCI, and then partially recovered, while the late components were lost without any recovery signals. All animals in group C showed no MEP waves excepting 2 rats had recovery signals. The size of the cord injuries area increased according to the dropping force and was correlated significantly with the amplitude of the largest peaks of scMEP 1 hour after SCI (r = -0.821).

CONCLUSIONThe scMEP changes after SCI are correlated with the injury forces and the pathological changes in the cord, which indicates that scMEP can be used as an objective index for the cord functional monitoring.

Animals ; Electric Stimulation ; Evoked Potentials, Motor ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; pathology ; physiopathology

OBJECTIVEThe aim of this study is to investigate the changes of hard tissue profile in anterior-posterior and vertical direction in bimaxillary protrusion patients after orthodontic treatment.

METHODSA total of 24 bimaxillary protrusion patients (male 8, female 16), aged from 11.2 to 29.0 (average 16.9 years old), were selected to be treated with standard edgewise technique consisted of 4 first premolars extraction. Cephalometrics were taken before and after treatment. The changes of hard tissue profile were studied using the computer-aid X-ray cephalometric analysis.

RESULTS1. The length of the maxillary and the mandible increased significantly, but the anterior-posterior relationship of the maxillary and the mandible did not change significantly. 2. The anterior and posterior facial height increased significantly, but the ratio of anterior and posterior facial height and the angle of MP-FH which reflected the inclination of the mandible plane did not change significantly. 3. The height of the upper and lower first molar increased significantly along with the increase of the anterior and posterior facial height. 4. The height of the upper incisors increased significantly, but the height of the lower incisors decreased significantly.

CONCLUSIONThe anchorage in anterior-posterior and vertical direction were controlled preferably, the patients did not manifest disadvantageous vertical growth trend.

Adolescent ; Adult ; Cephalometry ; Child ; Facial Bones ; pathology ; Female ; Humans ; Male ; Orthodontic Wires ; Orthodontics, Corrective ; Prognathism ; therapy ; Skull ; pathology

OBJECTIVESTo further investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.

METHODS25 normal control rats, 25 cirrhotic rats, 20 cirrhotic rats received saline, and 20 cirrhotic rats treated with cisapride were included in the study. All animals were assessed with many variables including bacterial and endotoxin translocation, IBO, intestinal transit and permeability.

RESULTSBacterial translocation was found in 48%(12/25) cirrhotic rats and none of control rats. Among the 20 rats with IBO, there were 11 rats with bacterial translocation (BT) while only one rats occurred BT out of the 5 rats without IBO. Cirrhotic rats with IBO had a significantly higher rate of endotoxin translocation, higher intestinal permeability and longer intestinal transit than those without IBO. BT of a specific organism was always associated with IBO of that organism. Compared with the placebo group, cisapride-treated rats had lower rates of bacterial and endotoxin translocation and IBO, which had close relationship with shorter intestinal transit and lower permeability.

CONCLUSIONEndotoxin and bacterial translocation in cirrhotic rats may be the result of IBO and higher permeability. IBO may be the result of longer transit. Cisapride which can accelerate intestinal transit and improve intestinal permeability is helpful in preventing and treating intestinal bacterial and endotoxin translocation.

Animals ; Bacterial Translocation ; drug effects ; Biological Transport ; Cisapride ; pharmacology ; Endotoxins ; metabolism ; Liver Cirrhosis, Experimental ; microbiology ; Male ; Permeability ; Rats ; Rats, Sprague-Dawley

The present study aimed to clone and express the EmLDH gene of Echinococcus multilocularis in Qinghai Province,identifying immunogenicity of EmLDH recombinant protein and evaluating its immune diagnostic value preliminarily.EmLDH genes were cloned by RT-PCR technology and linked into pET15b vector.Recombinant expression pET15b-EmLDH vectors were constructed and transformed into E.coli Rosetta (DE3) competent cells.Recombinant proteins were induced and expressed.Expression forms of recombinant proteins were detected by SDS-PAGE.Recombinant proteins were purified by affinity chromatography of Ni-IDA resin.Immunogenicity of recombinant proteins was identified by Western blotting.Serum samples from patients with alveolar echinococcosis (57 cases),cystic echinococcosis (33 cases),and healthy persons (50 cases) were examined by ELISA,which evaluated preliminarily immune diagnosis effect of EmLDH recombinant proteins.Results showed that EmLDH gene was cloned successfully and the recombinant proteins were expressed and purified.Results of Western blotting showed EmLDH recombinant proteins were recognised by serum samples from patients with alveolar echinococcosis and cystic echinococcosis,but not by serum samples from healthy persons.Results of ELISA showed that diagnostic sensitivities of EmLDH recombinant protein reacted with serum samples from patients with alveolar echinococcosis and cystic echinococcosis were 84.21％ and 84.85 ％ respectively.EmLDH recombinant proteins of Echinococcus multilocularis have high immunogenicity and good immune diagnostic value for echinococcosis.

OBJECTIVETo probe into a better therapeutic method for functional constipation.

METHODSNinety-five cases of functional constipation were randomly divided into deep puncture at ST 25 group (48 cases), shallow puncture at ST 25 group (24 cases) and medication group (23 cases). In deep puncture at ST 25 group, Tianshu (ST 25) was punctured deeply to the peritoneum, with electric stimulation. In shallow puncture at ST 25 group, Tianshu (ST 25) was punctured shallowly, 5 mm beneath the skin, with electric stimulation. In medication group, Duphalac was administered orally. These cases were treated continuously for 4 weeks in 3 groups and followed up for 6 months. It was to observe the numbers of person who had defecation 4 times a week, difference in weekly defecation frequency and the difference in the Cleveland Clinic Score (CCS).

RESULTSIn deep puncture at ST 25 group, the frequency of weekly defecation and the numbers of person who had defecation 4 times a week increased and CCS decreased, which were similar to the efficacy in shallow puncture at ST 25 group (all P > 0.05). But the efficacy of both ST 25 groups was superior to that in medication group (both P < 0.05). In comparison, the deep puncture at ST 25 group acted more quickly than either shallow puncture at ST 25 group or medication group and its efficacy remained much longer.

CONCLUSIONThe deep puncture at ST 25 with electric stimulation presents similar efficacy on functional constipation as shallow puncture at ST 25, but it acts more quickly than shallow puncture at ST 25, both of them are more advantageous than medication and the long-term efficacy is better.

Acupuncture Points ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To study the sensitivity and specificity of a double monoclonal antibody sandwich enzyme-linked immunosorbent assay (DMcAbS-ELISA)for the detection of F1 antigen of Yersinia pestis (Y.pestis).Methods Viscera (viz.liver and spleen)specimens of infected mice with virulent Y.pestis and negative control mice were detected by bacteriological test,DMcAbS-ELISA and reverse indirect hemagglutination assay (RIHA) for the F1 antigen.Results The 225 control specimens were all negative tested by plague bacteriology testing,DMcAbS-ELISA and RIHA.A total of 308 plague-infected mouse organ specimens were tested,and the positive detection rate was 92.21％ (284/308),90.91％(280/308) and 89.61％ (276/308),respectively,with germiculture,DMcAbS-ELISA and RIHA,and the difference was not statistically significant(x2=5.65,P＞0.05).The coincidence rate of DMcAbS-ELISA and bacterial culture was 97.00％[(274+243)/533],Kappa =0.940;RIHA in line with the rate was 99.25％[(276+253)/533],Kappa =0.985.Authenticity comparison of F1 antigen detection in viscera specimens:sensitivity,specificity,positive predictive value,negative predictive value,adjusted agreement and Youden's index was 96.48％(274/284),97.59％(243/249),97.86％ (274/280),96.05 ％(243/253),96.99％[1/4×(274/280+274/284+243/253+243/249)]and 0.9407,respectively,for DMcAbS-ELISA and 96.13％(273/284),98.80％(246/249),98.91％(273/276),95.72％(246/257),97.39％[1/4×(273/276+273/284+246/257±246/249)]and 0.9492,respectively,for RIHA.The detection sensitivity of DMcAbS-ELISA and RIHA was 2.7×104 cfu/ml and 2.2×105 cfu/ml,for Y.pestis,respectively,and was 10 μg/L for F1 antigen.Conclusions DMcAbS-ELISA assay is a sensitive,specific,simple and fast method for detection of the F1 antigen,and it has a potential application value in rapid diagnosis of plague.

As to terrible triad injury of elbow joint, because of the complexity of the surrounding anatomy of the elbow joint, the limited biomechanical study, the difficult treatments, and the poor prognosis, the choice of the treatment scheme is still controversial. This article summarizes the anatomy, injury mechanism, surgical treatment and prognosis of elbow joint by retrieving a large number of references. Currently, there are various surgical treatments for triad injury of elbow joint, but due to individual differences, the clinical efficacy is mostly uncertain. In the treatment of triad injury of elbow joint, most orthopedics doctors still choose the lateral surgical approach to fix the coronary fracture, repair or replace the radial head, routinely repair the lateral collateral ligament, and not recommend to repair the medial collateral ligament, so it still remains to be discussed.

Objective To develop a supporting pole for medical detachable camouflage net at field conditions to enhance the timeliness of battlefield camouflage of deployable units such as medical tent.Methods The pole had a dumbbell-shaped,hollow and columnar structure,which was composed of a base,a pole body and a terminal disc.The pole body consisted of internal and external parts.The external part had a vertical opening at its top and screw thread at its side wall,which was equipped with a binding bolt.The terminal disc had a circular structure and a 20 cm outer diameter,which was fixed 10 cm under the top of the internal pole to support the net.The base had a center hole and a sleeve to hold the lower part of the external pole to immobilize the supporting pole.Results The supporting pole decreased the deployment time of the camouflage net from 5 min to 2 min and the withdrawal time from 4 min to 1.5 min,and enhanced the timeliness of all-element deployment of mobile medical unit during field practical training.Conclusion The supporting pole meets the tactical requirements for deployment,storage,transport and robustness,and thus is worthy promoting in medical unit.