Biocompatibility of extracellular matrix hydrogel with human iPSCs differentiated cardiomyocytes / 中华心血管病杂志

Objective: To observe the biocompatibility of porcine omental derived extracellular matrix (ECM) hydrogel with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and the feasibility of ECM hydrogel as a delivery vector of cell transplantation. Methods: A series of chemical, physical and enzymatic methods were applied to acellularize the porcine omentum. Subsequently, the extracted ECM was prepared into thermosensitive hydrogel. The biochemical composition of the hydrogel was identified by histological staining. The microstructure was observed by scanning electron microscopy. The hydrogel was then injected into the myocardium of mice to observe its in situ gelation ability. Differentiation of human induced pluripotent stem cells into cardiomyocytes was achieved by small molecule induction, and then the obtained hiPSC-CMs were cultured. hiPSC-CMs cultured onto the prepared hydrogel were defined as the hydrogel group, while conventionally cultured hiPSC-CMs were defined as the control group. Cardiomyocyte viability and growth patterns were detected using live/dead staining, CCK-8 and phalloidin staining. Immunofluorescence staining and Western blot of cardiomyocytes were used to determine the survival and phenotypic maintenance markers of cardiomyocytes in materials. Results: The results of HE staining, oil red O staining and DAPI fluorescence staining showed that there was no significant cell debris, nucleus and lipid residue in the prepared ECM hydrogel. The Sirius red staining and Alcian blue staining showed that the hydrogel retained collagen and glycolaminoglycan, which were the main components of ECM. The prepared hydrogel behaves as a viscous liquid at 4 ℃ and as a gel state at 37 ℃. Scanning electron microscope results showed that the microstructure of the hydrogel was composed of irregular fibers and pores of different sizes. Under the guidance of ultrasound, the prepared ECM hydrogel could be successfully injected into the myocardium of mice. Immediately after the injection, the hyperechoic signal could be observed under ultrasound, suggesting that the hydrogel remained in the myocardium. HE staining of myocardial tissue evidenced that there was lump of gel in the injection area. The differentiated hiPSC-CMs were co-cultured with the prepared ECM hydrogel, and the results of live/dead staining showed that most of the hiPSC-CMs in the hydrogel group and the control group were alive, dead cells were scanty. The results of CCK-8 test showed that the absorbance values of the two groups were similar (P>0.05). The results of phalloidin staining showed that hiPSC-CMs could extend normally when co-cultured with ECM hydrogel. The cell morphology of the hydrogel group was similar with that of the control group, and there was no statistically significant difference in the F-actin coverage area per cell between the two groups (P>0.05). Immunofluorescence staining of cardiomyocyte markers showed that there was no significant difference in the coverage area of α-actinin and connexin-43 (Cx-43) per field between the hydrogel group and the control group (both P>0.05), the quantitative results of DAPI staining showed that there was no statistically significant difference in the number of cells between the two groups (P>0.05). Meanwhile, the results of Western blot showed that the expression levels of α-actinin and Cx-43 in cardiomyocytes in the hydrogel group were similar as those in the control group (both P>0.05). Conclusions: These results show that preparation of the ECM hydrogel from porcine omentum is successful. The hydrogel has good biocompatibility and no obvious cytotoxicity. Besides, the hydrogel can support the survival of hiPSC-CMs in vitro and maintain its phenotype. These properties make it a promising injectable cardiac tissue engineering material.

Effect and Mechanism of Buyang Huanwu Tang on Atherosclerotic Plaque Based on Treg Cells / 中国实验方剂学杂志

Objective::To explore the effect and mechanisms of Buyang Huanwu Tang (BYHWT) on atherosclerotic plaque based on regulatory T cells (Treg) and inflammation. Method::Totally 50 ApoE knockout(ApoE-/-)mice aged 8 weeks were randomly divided into model group, low, medium, high-dose BYHWT groups, positive control group, and C57/BL mice were taken as control group. The model group and the BYHWT group were given high-fat diet for 12 weeks, while the control group was given normal diet. After successful modeling, BYHWT groups were given drugs (5, 10, 20 g·kg-1) through intragastric administration, the positive control group was given rapamycin (4 mg·kg-1), while the control group and the model group were given equal doses normal saline through intragastric administration for 4 weeks. Four weeks later, the mice were sacrificed. Manufactured paraffin sections were prepared for the aortic sinus of the heart. The plaque area was evaluated by hematoxylin and eosin (HE) staining, and the number of Treg cells in immunohistochemical staining plaque was detected. Blood was collected from eye canthus of mice, the expression of inflammatory factors in peripheral blood was detected by enzyme-linked immunosorbent assay (ELISA), and the forkhead box P3 (Foxp3) gene expression in peripheral blood was detected by Real-time fluorescent quantitative polymerase chain reaction (PCR). Result::Compared with the control group, the area of atherosclerotic plaques in the model group was significantly increased (P<0.01), the contents of serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were significantly increased (P<0.05), and transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) were significantly decreased (P<0.05), and the peripheral blood Fxop3 mRNA expression was decreased (P<0.05). Compared with the model group, the plaque areas in middle-dose and high-dose BYHWT groups were significantly reduced(P<0.05), the peripheral blood TNF-α and IL-6 contents were decreased (P<0.01), the TGF-β and IL-10 expressions were increased in the high-dose group (P<0.05, P<0.01), the number of Treg cells in the plaque was increased in the high-dose group (P<0.01), and the peripheral blood Fxop3 mRNA expression was increased in each BYHWT group (P<0.01). Conclusion::BYHWT has an anti-atherosclerosis effect, which may be related to the increase of the number of Treg cells and thereby inhibiting the inflammatory response in vivo.

Application effect of cardiac remote real-time monitoring system in pre-hospital rescue / 心血管康复医学杂志

Objective: To study application effect of cardiac remote real-time monitoring system in pre-hospital rescue. Methods: A total of 400 patients with coronary heart disease complicated arrhythmia treated in our hospital were selected. They were chronologically and equally divided into routine monitoring group (received routine bedside 12-lead ECG examination and ECG results were checked regularly by physicians and nurses during bedtime) and remote monitoring group (wore cardiac remote monitoring alert reporter, results were recorded by multi-channel simultaneously and auto-delivery mode was activated). Abnormal results recorded by real-time monitoring and time to identify patient's abnormal condition were compared between two groups, and application effect was evaluated. Results: There were no significant difference in percentages of ventricular tachycardia, supraventricular tachycardia, paroxysmal atrial fibrillation, atrioventricular block, bundle branch block, premature ventricular contraction Lown grade I～II and≥grade III between two groups, P＞0. 05 all. Compared with routine monitoring group, there was significant rise in percentage of patient's abnormal condition identified within 10min (38％ vs. 52％), and significant reductions in percentages of patient's abnormal condition identified within 10～30min (44％ vs. 28％) in remote monitoring group, P＜0. 05 all. Conclusion: Cardiac remote real-time monitoring system possesses the advantages of rapid diagnosis, long transmission distance and simple operation, etc., which is worth extending.

Research progress in the structure and function of dengue virus non-structural 1 protein / 病毒学报

Dengue virus (DENV) is a re-emerging disease transmitted by the Aedes mosquitoes and has become a major public health problem in southern China. Currently, no antiviral drug or effective vaccine exist to control this disease. The chimeric DENV structural protein vaccine cannot elicit balanced levels of protective immunity to each of the four viral serotypes; therefore, non-structural protein components may be required to construct an effective DENV vaccine. The Dengue virus non-structural 1 (DENV NS1) protein plays a critical role in viral pathogenesis and protective immunity. Therefore, immunity to Dengue 1-4 NS1 subtypes may be crucial for the prevention of severe disease. This review attempts to provide an overview about the structure and function of DENV NS1.

The protective role of interleukin-6 monoclonal antibody on experimental autoimmune myocarditis and its mechanism / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of IL-6 mAb on experimental autoimmune myocarditis (EAM) in rats, and search the mechanism of the role of IL-6, helper T cells 17 (Th17) and regulative T cells (Treg) in EAM pathogenesis.</p><p><b>METHODS</b>Thirty-four Lewis rats were divided into three groups randomly, i.e. control group (n = 6), EAM group (n = 12), and IL-6 mAb intervention group (n = 16). Rats in EAM group and IL-6 mAb intervention group were injected intracutaneously with myosin to establish EAM model. Rats in IL-6 mAb intervention group were injected intraperitoneally with 1 mg IL-6 mAb on 1st, 7th to 20th day after cardiac myosin immune injection. Myocardial inflammation was examined by HE stain, Masson stain, and TdT assay (TUNEL reaction) on 21st and 84th day after IL-6 mAb therapy in order to assess the therapeutic role. Spleen cells were analyzed by flow cytometry to illustrate Th17 and Treg cells? number and function. The serum concentration of IL-6, IL-10, IL-17, and TGF-beta in each group was measured by ELISA, concentration of STAT3, RORgammat, and Foxp3 mRNA in each group was determined with RT-PCR. Spleen cells derived from EAM were stimulated by IL-6 mAb in vitro, and the concentration of IL-10, IL-17 and TGF-beta was measured by ELISA.</p><p><b>RESULTS</b>Inflammation score, fibrosis score, and apoptosis index in IL-6 mAb intervention group were significantly decreased as compared with those in EAM group (P < 0.01). The number of Th17 and Treg cells in EAM group on the 21st day (experimental acute peak stage) were increased, and those in intervention group on the 21st day were significantly inhibited (P < 0.01). The concentration of serum IL-6, IL-10, IL-17 and TGF-beta in intervention group on the 21st day was decreased dramatically in comparison with that in EAM group on the same day (P < 0.01). The levels of peripheral blood STAT3, RORgammat, Foxp3 mRNA in intervention group on the 21st day was decreased significantly as compared with that in EAM group (P < 0.01). The expression of IL-10, IL-17 and TGF-beta was increased significantly (P < 0.01) by stimulation of IL-6 mAb on spleen cells derived from EAM in vitro.</p><p><b>CONCLUSIONS</b>IL-6 mAb could neutralize IL-6, and ameliorate myocarditis and reduce heart autoimmune responses. IL-6 mAb has significantly protective effects on EAM by suppressing Th17 and Treg cells.</p>

Study of L-NAME treatment on experimental autoimmune myocarditis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of N-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NOS, on experimental autoimmune myocarditis (EAM) in Balb/C mice and discuss the therapeutic mechanism induced by apoptosis.</p><p><b>METHODS</b>Thirty male Balb/C mice were divided into normal control group, model control group and experimental group randomly (n = 10). Model control group and experimental group were created into EAM by injection of porcine cardiac myosin subcutaneously in double groin and axilla and pertussis toxin intraperitoneally on day 0 and 7 respectively. Model control group was intraperitoneally administered 5 mg/(kg x day) of physiological saline after infective myosin and pertussis toxin. Experimental group was intraperitoneally given 5 mg/(kg x day) of L-NAME on day 1-21. The hearts and blood were processed after sacrificed on day 21. Cardiac inflammation score was measured by HE staining. Heart weight / body weight (HW/BW), serum nitric oxide (NO) level, activity of induced nitric oxide synthase (iNOS) and mRNA expression of iNOS in heart were measured in each group. Degree of heart apoptosis were evaluated by cardiac apoptotic index through TUNEL, immunohistochemical examination and real time PCR of Caspase-3, Caspase-8 and Caspase-9.</p><p><b>RESULTS</b>Compared with normal control group, cardiac inflammation score, HW/BW level of NO and activity of iNOS, mRNA expression of iNOS, the levels of mRNA and protein of Caspase-3, Caspase-8 and Caspase-9 and cardiac apoptotic index were significantly higher (P < 0.01) in model control group, and those of model control group were higher than those of experimental group (P < 0.01). HW/BW was only a little elevation in model control group compared with that in the experiment group (P < 0.05).</p><p><b>CONCLUSION</b>The development of EAM is related with the NO catalyzed by iNOS. L-NAME protects cardiac myocyte via suppressing the activity of iNOS and further decreased production of NO in EAM. The mechanism might be that L-NAME alleviated myocardial inflammation through inhibited the apoptosis of cardiac myocyte.</p>

Clinical significance of ultrasound-assisted drawings in location of thyroid micronodule / 中华医学超声杂志（电子版）

Objective To explore the clinical significance of preoperative ultrasound-assisted drawings in location of thyroid micronodule resection .Methods A total of 88 patients ( 137 nodules ) who underwent thyroid micro-nodule resection were enrolled in the prospective randomized controlled study .All patients were randomly divided into two groups:46 patients (68 micronodules) in experimental group and 42 patients (69 micronodules) in control group.Inclusion criteria:the maximum diameter of nodule≤1.0 cm. Preoperative thyroid ultrasound was conducted .Patients in experimental group also underwent ultrasound-assisted location of thyroid micronodule .Results The diagnostic accuracy rate of US was 81.82%(69/88) in all patients .With the help of ultrasound-assisted location , all nodules in experimental group were found quickly and accurately.The resection rate of experimental group was 100%(46/46).Whereas 4 nodules (in 4 patients ) were missed in control group with a resection rate of 90.5% (38/42).The postoperative US examinations after 3 months showed that all nodules were completely removed in experimental group while 4 nodules retained in control group .Conclusions Preoperative ultrasound-assisted drawings in location of thyroid micronodule plays an important role in thyroid nodule resection .It is great value of clinical application .

Application of ultrasound-guided drawing location of thyroid micronodule / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the application value of preoperative ultrasound-guided drawing for locating thyroid micronodule in surgery.</p><p><b>METHODS</b>A total of 88 patients (with 137 thyroid micronodules) who underwent thyroid surgery was included in the prospective study. Preoperative thyroid ultrasound was conducted in all patients. Select criteria: the maximum diameter of nodule ≤ 1 cm. All patients were randomly divided into two groups: 46 patients (68 micronoduls) in experimental group with ultrasound-guided drawing location of thyroid micronodule and 42 patients (69 micronoduls) in control group without ultrasound-guided drawing location of thyroid micronodule.</p><p><b>RESULTS</b>All thyroid micronodules of experimental group were found quickly and accurately in surgery, and 4 micronodules in 4 patients of control group were not found in surgery. US examinations 3 months after surgery showed that all micronodules in experimental group were completely removed and 4 micronodules in control group retained.</p><p><b>CONCLUSION</b>Ultrasound-guided drawing is a useful technique for locating and searching accurately thyroid micronodule in surgery.</p>

Analysis of genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex(ADC) , so as to research the amino acid variability and the pathogenesis of ADC.</p><p><b>METHODS</b>The nef gene was amplified with PCR from genomic DNA which was extracted from spleen and different brain tissues(basal ganglia, frontal gray matter, meninges, temporal lobe)of a patient who died of ADC. PCR products were cloned into the pMD19-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced and confirmed with BLAST. HIV-1 nef sequences were processed with BioEdit and MEGA4 to do Neighbor-Joining tree, p-Distances, and values of ds/dn.</p><p><b>RESULTS</b>The samples were all identified as HIV-1 B and genetic variation exists in HIV-1 nef gene isolated from different tissues compared with HXB2. In addition,part of the changes were different between periphery and brain.</p><p><b>CONCLUSION</b>Variations exist in the HIV-1 nef gene extracted from the ADC patient and the variations from peripheral and central nerve tissues were different,these variations may change the function of Nef,and it needs more research.</p>

Establishment of human cardiac C protein induced experimental autoimmune myocarditis model in rat / 中华心血管病杂志

<p><b>OBJECTIVE</b>To construct the recombinant plasmid of human cardiac C protein (CCP) peptide with immunogenicity and to express, purification and renature fusion protein. The fusion protein was injected to Lewis rats to establish experimental autoimmune myocarditis (EAM) model.</p><p><b>METHODS</b>Total RNA was extracted from human heart and used as the template for reverse transcriptase-directed cDNA synthesis. The cDNA was then amplified by polymerase chain reaction (PCR) using oligonucleotide primers specific for CCP peptide with immunogenicity. Subsequently, the purified CCP peptide gene was cloned into PEASY-T1 vector and the ligated product was identified by PCR and DNA sequence analysis. Then the CCP target gene of positive clone was inserted into the pQE30, a prokaryotic expression vector, and the inserting plasmid was transformed into Escherichia coli. host M15. The positive clone extracted from the bacterium liquid was sieved by insertional inactivation sieve method and identified by PCR of bacterium liquid, CCP immunological peptide was purified and renatured in semipermeable membrane. EAM model in Lewis rats was induced by injection of mixture of 100 µg CCP fusion protein immunological peptide and 2.5 g/L completed Freund adjuvant from two double foot pad and subsequent abdominal injection of 0.5 µg pertussis toxin. Two, four, six, and eight weeks after immunization, hemodynamic evaluation was made and hearts underwent histological examination.</p><p><b>RESULTS</b>The DNA sequence analysis for cloning vector extraction revealed that the CCP target gene was cloned into pQE30 exactly. The DNA of 1000 bp length was obtained by PCR examination of bacterium liquid with transformation of express recombinants which were consistent with the expected size. Purified fusion protein in vertical slab gel electrophoresis showed 35 000 as expected. The recombinant CCP fusion protein existed in inclusion bodies of E. coli and amounted to 80% - 90% of the total protein. Hemodynamic and histological evaluations showed typical acute inflammatory responses at 2 weeks, subacute inflammatory and fibrosis changes at 4 weeks after injection, and signs of chronic dilated cardiomyopathy at 6 weeks post injection.</p><p><b>CONCLUSION</b>Combination of gene clone technique and histidine tag protein purification technique can be used to synthesize human cardiac C protein to induce EAM model in Lewis rat.</p>

Prognostic value of the lymph node ratio in stage III colorectal cancer / 癌症

The nodal stage of colorectal cancer is based on the number of positive nodes. It is inevitably affected by the number of removed lymph nodes, but lymph node ratio can be unaffected. We investigated the value of lymph node ratio in stage III colorectal cancer in this study. The clinicopathologic factors and follow-up data of 145 cases of stage III colorectal cancer between January 1998 and December 2008 were analyzed retrospectively. The Pearson and Spearman correlation analyses were used to determine the correlation coefficient, the Kaplan-Meier method was used to analyze survival, and the Cox proportional hazard regression model was used for multivariate analysis in forward stepwise regression. We found that lymph node ratio was not correlated with the number of removed lymph nodes (r = -0.154, P = 0.065), but it was positively correlated with the number of positive lymph nodes (r = 0.739, P < 0.001) and N stage (r = 0.695, P < 0.001). Kaplan-Meier survival analysis revealed that tumor configuration, intestinal obstruction, serum carcinoembryonic antigen (CEA) concentration, T stage, N stage, and lymph node ratio were associated with disease-free survival of patients with stage III colorectal cancer (P < 0.05). Multivariate analysis showed that serum CEA concentration, T stage, and lymph node ratio were prognostic factors for disease-free survival (P < 0.05), whereas N stage failed to achieve significance (P = 0.664). We confirmed that lymph node ratio was a prognostic factor in stage III colorectal cancer and had a better prognostic value than did N stage.

Characteristic ultrasonographic features of the encapsulated papillary thyroid carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the ultrasound (US) features of the encapsulated papillary thyroid carcinoma (EPTC).</p><p><b>METHODS</b>Based on ultrasonographic features including shape, size, border, echogenicity, hypoechoic halo and microcalcification, 33 cases of EPTC were classified into two groups: 21 cases in irregular shape group and 12 cases in spherical or oval shape group.</p><p><b>RESULTS</b>EPTC in the irregular shape group showed some ultrasonographic features including jagged border, irregular tumor shape and marked hypoechogenicity, while the ultrasonographic features of EPTC in the spherical or oval shape group included smooth border, regular shape, isoechogenicity and hypoechoic halo. Hypoechoic halo and isoechogenicity were found more frequently in EPTC of spherical or oval shape group than those in EPTC of irregular shape group. The size of EPTC in the spherical or oval group was commonly larger than that of EPTC in the irregular shape group.</p><p><b>CONCLUSIONS</b>The findings indicate that EPTC have some ultrasonographic features similar to benign follicular thyroid tumors.</p>

Digital visualization construction and research on Jiuwei (CV 15) / 中国针灸

<p><b>OBJECTIVE</b>To establish the digitized visible human with Jiuwei (CV 15) involved.</p><p><b>METHODS</b>With the virtual Chinese human (VCH) datasets and three-dimensional modeling software adopted, the knowledge on acupuncture and moxibustion as well as acupoint anatomy combined and the computer image processing software applied, the visualized browser software of Jiuwei (CV 15) was established.</p><p><b>RESULTS</b>By establishing the interactive three-dimensional visualization browser software of acupoints, the location of Jiuwei (CV 15) was enabled to be expressed directly from all the levels, as well as the main adjacent tissue structure. It was suggested that Jiuwei (CV 15) should be punctured obliquely downward to avoid injuring the vital organs such as the heart and liver, and the safe depth of insertion should be 1.0-1.5 cm.</p><p><b>CONCLUSION</b>The technology of digital visible human enables the three-dimensional expression of acupoint, which can be the platform of the digital teaching pattern. The research on the angle and depth of needling insertion at acupoint can be conducted in combination with teaching materials.</p>

Clinical evaluation of periapical endodontic surgery for endodontic failure / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical outcome of periapical endodontic surgery for teeth that can't be treated by nonsurgical endodontic methods.</p><p><b>METHODS</b>Sixty-two affected teeth were chosen for surgical endodontic treatment, of which 31 teeth underwent periapical curettage and the others were treated by root-end resection, retrograde preparation and filling. A radiography was taken immediately after surgery and was compared with those taken at 12 and 24 months. The results of two groups were analyzed using the chi2 test.</p><p><b>RESULTS</b>The success rate for retrograde filling was higher (85% after 12 months, 88% after 24 months) compared with that of periapical curettage (52% after 12 months, 45% after 24 months). The difference in success rate between the two groups was statistically significant.</p><p><b>CONCLUSIONS</b>Ultrasonic root-end preparation and retrograde filling is a good choice of treatment when the teeth can't be treated appropriately by nonsurgical treatment.</p>

Study an E protein epitopes and primary identification of main yellow virus / 中华流行病学杂志

Objective To analysis the E protein epitopes of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus and to distinguish the shared or specific epitopes among them. Methods Bioinformatic software DNAStar was used to analyze the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E prtein amino acid sequences. The influence of secondary structure was also considered. Based on the bio-informatic analysis of E protein epitopes, 6 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-c2x. The vectors was then transferred into E.coli BL21 (DE3) and Rosetta (DE3). Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of gene segments and SDS-PAGE were used identify the expression proteins. The antigenieity was tested, using Western blot. Results 15 shared epitopes and 47 specific epitopes were forecasted by bioinformatic analysis, and 6 specific epitopes from dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E protein were expressed in E.coli successfully. Two specific antigenic determinant from dengue virus type 1 and dengue virus type 2 were confirmed using Western blot, while the others epitopes shown no antigenic reaction property. Conclusion Two specific antigenic determinant were confirmed, under Western blot.

Detection and typing of dengue virus using polymerase chain reaction and microwell plate hybridization / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a specific, sensitive and practicable method for detection and typing of dengue virus.</p><p><b>METHODS</b>Based on the genomic sequence analysis of dengue virus types 1-4, 4 pairs of primers were designed. The specific capture probes of dengue virus types 1-4 were amplified using RT-PCR, cloned and sequenced before using them for precoating the microwell plate. The samples were amplified using biotin-labeled forward primer and reverse primer, and microwell plate hybridization was carried out for detection and typing of dengue virus types 1-4.</p><p><b>RESULTS</b>The absorbance of the positive samples were higher than 0.5, while the average absorbance of the negative samples was lower than 0.1, with the S/N higher than 10.</p><p><b>CONCLUSION</b>The method of PCR-ELISA we established for early detection and typing of all 4 dengue viruses seretypes.</p>

Clinical management of open apices teeth with mineral trioxide aggregate (MTA) as apical barrier in adults / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical effect of treatment of open apices teeth with mineral trioxide aggregate (MTA) compared with apexification using Vitapex in the adult.</p><p><b>METHODS</b>The root canals of 41 anterior teeth and premolars with single canal and open apices in adults were cleaned and shaped, and then disinfected with calcium hydroxide. The apical root canals of 21 teeth in experimental group were filled with MTA to create a 3-5 mm apical barrier. The remainder of the canals was filled with AH plus and Obtura II gutta-percha. The canals of 20 teeth in control were treated with Vitapex. The root canals were then filled with Obtura II gutta-percha when the apical barrier could be detected. The visit times, period and result of the treatment were recorded for all cases.</p><p><b>RESULTS</b>Good result was achieved in the most cases in the experimental group. The postoperative X-ray films showed that the canals of 15 teeth were obturated well. 6 teeth showed over-filling by 0.5-2 mm. The recalled patients declared their teeth to be asymptomatic except one. The recall radiographs indicated that the apical radiolucent areas of the teeth with pre-existing apical lesion decreased apparently or disappeared completely, except one tooth with large apical radiolucency. No new radiolucency was found around the roots. In the control group, the apexification of 11 teeth succeeded, and 9 failed. The average visit times was 3.5 in experimental group and 6.0 in control. The average period of the whole endodontic treatment was 11.8 days in experimental group and 306.8 in control.</p><p><b>CONCLUSION</b>MTA is more effective and quicker than apexification in treatment of teeth with open apices in adults.</p>

An Aldose Reductase Inhibitor Screening Model Constructed by Transfection of pSNAV-AR into HEK293 Cells / 中国生物工程杂志

Objective:Aldose reductase,involved in the pathogenesis of diabetic complications,was recombinated with an adeno associated virus vector pSNAV2.0,and it was transfected into human embryonic kidney 293(HEK 293)cells.The gene engineering produced AR would be used as a target protein to screen aldose reductase inhibitors.Restriction endonuclease digestion and ligation procedures were performed to construct the AR expression plasmid vector pSNAV-hAR.Methods:After confirmation the recombinant plasmid by PCR,restriction endonuclease digestion,and DNA sequencing,pSNAV-hAR was transfected into HEK293 cells.Western blot and immunofluorescence analysis were performed to detect the expression of AR and its enzyme activity.Results:The results of a series of analysis including AR activity assay,Western blot and immunofluorescence analysis shown the expressed protein mediated by the adeno associated virus vector transfecting HEK 293 cells,was functional AR.The traditional aldose reductase inhibitors,Sobinil and Zopolrestat,were used to test and verify the constructed cell model.Conclusion:The established AR expression model can be used in mechanismresearch of activation of polyol pathway on diabetic complications and screening potential aldose reductase inhibitors.

Application of modified TransFix-II technique in arthroscopic reconstruction of the anterior cruciate ligament using double tibial tunnel / 中国医学科学院学报

<p><b>OBJECTIVE</b>To describe a modified TransFix- 11 technique for arthroscopic reconstruction of anterior cruciate ligament (ACL) using double tibial tunnel.</p><p><b>METHODS</b>Twelve cases of ACL ruptures were reconstructed anatomically using modified TransFix- 11 technique. The double-looped semitendinosus and gracilis (DLSTG) tendon autograft was placed in a single femoral tunnel and double tibial tunnels to replace the anteromedial (AM) and posterolateral (PL) bundles of the original ACL. All the 12 patients underwent the same postoperative accelerated rehabilitation program.</p><p><b>RESULTS</b>Lachman test, anterior drawer test and pivot shift sign, and mcmurray test showed negative results in all cases. All patients regained normal range of motion of the knee and gait 4-6 weeks after operation, and 8 patients returned to the low risk sports at 10-12 weeks. One year after operation, 9 patients were followed: the International Knee Document Committee (IKDC) analysis revealed normal or near normal knees in 9 patients. The Tegner score revealed that 6 patients regained their pre-injury activity level. No patient had a significant femoral or tibial tunnels enlargement, as shown by follow-up CT scan.</p><p><b>CONCLUSION</b>Arthroscopic ACL reconstruction with double tibial tunnel using DLSTG and the technique of modified TransFix-II is reliable for replacement of the AM and PL bundles of the original ACL. The postoperative accelerated rehabilitation and the rapid restoration of the injured knee function can be expected.</p>

Development of multiplex reverse translation-polymerase chain reaction methods for detection of dengue virus type 1-4 and its application in clinical use / 中华流行病学杂志

<p><b>OBJECTIVE</b>To develop multiplex reverse translation-polymerase chain reaction (RT-PCR) method for detection of dengue virus type 1-4.</p><p><b>METHODS</b>Based on the genomes sequence analysis of dengue virus type 1-4, four-pair of primers were designed. The specificity of the primers was primarily tested by searching the GenBank DNA sequence database. The optimal reaction conditions of the multiplex RT-PCR were then established. The specificity of RT-PCR was tested using the homologous yellow fever virus and Japanese encephalitis virus. 30 serum samples of dengue virus from suspected sufferers in the prevalence of dengue virus in 2003 were detected using the methods we developed.</p><p><b>RESULTS</b>Positive segments about 295, 237, 118, 347 bp could be seen in the multiplex RT-PCR production of dengue virus type 1-4, respectively. There were no positive segments in the RT-PCR productions of Japanese encephalitis virus and yellow fever virus. 25 of the 30 serum samples showed dengue virus type 1 positive results, while the sequencing results suggesting the amplification sequence having a high homology with dengue virus type 1 strain Cambodia, GD14/97 and GD05/99 (97%, 97%, 98%, respective).</p><p><b>CONCLUSION</b>The method of multiplex RT-PCR we established could be used for early detection and identification of dengue virus type 1-4.</p>