AlM: To study the therapeutic effect of external-route microsurgery forrhegmatogenous retinal detachment.
METHODS: ln 55 patients ( 55 eyes ) with rhegmatogenous retinal detachment, drainage of subretinal fluid, examination of locating the holes, sclera cryotherapy, scleral buckling, and vitreous cavity injection of filtrated air were performed under surgical microscope.
RESULTS:The retinal reattachment occurred in 50 cases after the primary surgery. The final rate of reattachment was 91% during 6 - 12mo follow - up. The retinal reattachment occurred in 1 case ( recurrent retinal detachment) after the secondary surgery and in 4 cases ( recurrent retinal detachment ) after vitrectomy. The eyesight was improved with different degrees in 55 cases.CONCLUSlON: The external- route microsurgery for rhegmatogenous retinal detachment is simple, safe and effective.

Objective To examine mycological profile of eryptococcal meningitis in patients with non-acquired immune deficiency syndrome (AIDS) during treatment and follow-up so that to support clinical therapy. Methods Data of 28 cuhure-confirmed cryptoeoccal meningitis patients with non-AIDS were retrospectively analyzed. Fungat smear, count, culture and latex agglutination test of cerebrospinal fluid (CSF) were done during treatment and follow-up. Initial treatment included intravenous amphotericin B plus oral flucytosine or f;uconazole for at least 6 weeks, and consolidation treatment included oral fluconazole and (or) itraeonazole for at least 2 months. All 28 patients were cured. The data were analyzed by rank-sum test. Results The positive rate of CSF fungal smear was 92.9% before treatment and gradually decreased, and the fungal count was significantly reduced over time after treatment. While fungal smears of some patients were still positive after initial treatment. Fungal growth time in culture was gradually extended, and fungal culture turned to be negative in all patients after 2 weeks of treatment. The positive rate of latex agglutination test of CSF was 100%. Cryptococcal antigen titer decreased steadily after treatment, which was not correlated with the decrease of fungal count. Conclusion Mycological tests of patients with eryptococcal meningitis should be interpreted comprehensively during treatment, and result of each test should be specifically analyzed.

Objective To investigate the histological mechanism of the bone loss of castrated male rats and provied theoretical basis for clinical treatment.Methods 30 male rats aged 10 months were randomly divided into two groups as normal and castrated pathological groups.The animals were sacrificed after adminsration Heihudan (a TCM agent) for 12 weeks.L1 and L2 were chosen to make bone slices without decalificating and then diverse bone indexes and superficial indexes of bone morphological metrology were determined.ANOVA was used for data anlysis.Results Bone indexes in the pathological group were smaller than the normal one(P<0.05).The superficial indexes show:osteoclast index increased,osteogenesis index decreased,bone transformation became faster than before(P<0.05).Conclusion Reconstruction of negative balance and high bone transformation were the mechanisms of the bone loss of castrated male rats.The method of combining with drugs to promote bone formation,restrain bone absorption,reduce bone transformation seems to be useful for the treatment of senile male patients with osteoporosis.

Objective To explore the clinical application of percutaneous endoscopic gastrostomy in gastroin- testinal nutrition among critically ill patients.Methods Twenty-two patients were undertaken percutaneous endo- scopic gastrostomy under the lead of gastroscope.The puncturatio site was located in anterior wall of stomach.Results All the procedures were performed successfully for one time.Intraoperational blood pressure was very steady.At the same time,intraoperational SpO_2 of the patients all exceeced 97%.The bleeding amount and operation time were respectively (3.8?1.9) ml and (15.5?2.3) min.Severe complications such as gastrostoma,gastrocolic fistula, pneumoperitoneum,refluxing or aspiration of gastric juice,inhalant pneumonia didn't occur after the operation.Con- clusion Percutaneous endoscopic gastrostomy was safe and feasible among critically ill patients who needed gastroin- testinal nutrition because of its slight injury,little bleeding and shorter operational time.

BACKGROUNDPathophysiological processes, such as malignancy, can lead to the formation of stiffer tissue in lung cancers. Endobronchial ultrasound (EBUS) elastography is a novel technique for measuring tissue stiffness during EBUS-guided transbronchial needle aspiration (EBUS-TBNA). The current study was conducted to investigate the diagnostic value of EBUS elastography for mediastinal and hilar lymph node metastasis in lung cancers.

METHODSFrom January 2014 to January 2015, 40 patients suspected of lung cancer were enrolled, and a total of 68 lymph nodes were evaluated by EBUS-TBNA. EBUS-guided elastography of lymph nodes was performed prior to EBUS-TBNA. Standard EBUS characteristics were also described. Pathological determination of malignant or benign lymph nodes was used as the gold standard for this study. If EBUS-TBNA did not result in a formal pathological diagnosis of malignancy, patients were referred for a surgical procedure. Comparisons of elastography and standard EBUS characteristics were made between benign and malignant lymph nodes.

RESULTSElastography grading scores and strain ratios showed significant differences between benign and malignant lymph nodes (P = 0.000). The elastography strain ratio was more sensitive and specific for determining malignant lymph nodes than elastography grading score or standard EBUS criteria. The receiver operating characteristic curve for the elastography strain ratio showed an area under the curve of 0.933. The best cut-off point of the strain ratio for differentiating malignant from benign lymph nodes was 32.07. The elastography strain ratio had a sensitivity of 88.1%, the specificity of 80.8%, positive predictive value of 88.1%, and negative predictive value of 80.8% for distinguishing malignant from benign nodes. The overall accuracy of elastography strain ratio was 85.3%. The strain ratio of malignant and benign lymph nodes positively correlated with the elastography grading score (r = 0.561, P = 0.000).

CONCLUSIONSEBUS elastography can be effectively used to predict mediastinal and hilar lymph node metastases in lung cancer. This noninvasive technique may thus complement standard EBUS and help guide EBUS-TBNA procedures.

Aged ; Elasticity Imaging Techniques ; methods ; Endosonography ; methods ; Female ; Humans ; Lung Neoplasms ; diagnosis ; Lymph Nodes ; pathology ; Male ; Mediastinum ; pathology ; Middle Aged

OBJECTIVE: To explore the value of Wisconsin Card Sorting Test (WCST) in evaluating the recognition impairment after traumatic brain injury (TB1). METHODS: WCST and Wechsler Adult Intelligence Scale Revised in China (WAIS-RC) were adopted to assess the cognitive function in 186 patients with brain injury (experimental group) and 180 healthy volunteers (control group), respectively. The cognition between the experimental group and the control group was compared. The correlation between WCST and IQ was analyzed. The correlation between age, education level, the severity of TBI and the executive function were also analyzed. RESULTS: (1) The performances of WAIS-RC and WCST in the experimental group were significantly worse than that of the control group. (2) There were significantly negative correlation between the performance of total errors, percentage of preservative errors, percentage of random errors, response number on first category and IQ (P < 0.05). While there were significantly positive correlation between the categories completed, percentage of conceptual level and IQ (P < 0.05). (3) There was significantly negative correlation between percentage of conceptual level and the severity of TBI. While there were significantly positive correlation between percentage of preservative errors, percentage of random errors, response number on first category and the severity of TBI. CONCLUSION WCST could be an important method in evaluating cognition of patients with brain injury.
Adolescent ; Adult ; Aged ; Brain/pathology* ; Brain Injuries/psychology* ; Case-Control Studies ; Cognition Disorders/etiology* ; Executive Function/physiology* ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Neuropsychological Tests ; Predictive Value of Tests ; Psychiatric Status Rating Scales ; Psychometrics ; Reproducibility of Results ; Severity of Illness Index ; Task Performance and Analysis ; Wechsler Scales ; Young Adult

In order to investigate the special role of HOXB4 in expansion and self renewal of hematopoietic stem cells, the cDNA of HOXB4 was extracted and cloned from umbilical cord blood mononuclear cells by using RT-PCR. Then the eukaryotic expression bicistronic plasmid vector pIRES2-EGFP/HOXB4 was designed and constructed after cutting HOXB4 and pIRES2-EGFP respectively by restriction enzyme EcoRI and BamHI. The recombinant plasmid was delivered into competent cells of Escherichia coli. The successful construction of plasmid was confirmed by the identification of endonuclease cutting and sequencing. The results showed that the HOXB4 cDNA was cloned successfully from umbilical cord blood mononuclear cells and the recombinant eukaryotic expression bicistronic plasmid vector was constructed, and then introduced it into 293T cells successfully. It is concluded that a pIRES2-EGFP/HoxB1 eukaryotic expression bicistronic plasmid vector has been constructed successfully, which results provide a useful material basis for exploration of HoxB4 function in the proliferation and differentiation of hematopoietic cells.
Cell Line ; Cloning, Molecular ; Gene Expression ; Genes, Homeobox ; Genetic Vectors ; Humans ; Plasmids ; Transfection

OBJECTIVETo inhibit HBV core antigen gene expression with plasmid-based RNAi.

METHODSThe shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR.

RESULTSHBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR.

CONCLUSIONEffective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.

Cell Line ; Embryo, Mammalian ; Gene Expression Regulation, Viral ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Humans ; Kidney ; cytology ; virology ; Plasmids ; Polymerase Chain Reaction ; RNA Interference ; physiology ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo construct a cDNA library from human liver tissue of cirrhosis.

METHODSThe total RNA from human liver tissue of cirrhosis was extracted using Trizol method, and the mRNA was purified using mRNA purification kit. SMART technique and CDSIII/3' primer were used for first-strand cDNA synthesis. Long distance PCR was then used to synthesize the double-strand cDNA that was then digested by proteinase K and Sfi I, and was fractionated by CHOMA SPIN-400 column. The cDNA fragments longer than 0.4 kb were collected and ligated to lambdaTripl Ex2 vector. Then lambda-phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries was strictly checked by conventional titer determination. Eleven plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector.

RESULTSThe titers of unamplifed and amplified libraries were 1.03 x 10(6) pfu/ml and 1.36 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 97.24 % in unamplified library and 99.02 % in amplified library. The lengths of the inserts were 1.02 kb in average (36.36 % 1 approximately equals 2 kb and 63.64 % 0.5 approximately equals 1.0 kb).

CONCLUSIONA high quality cDNA library from human liver tissue of cirrhosis was constructed successfully, which can be used for screening and cloning new special genes associated with the occurrence of cirrhosis.

DNA, Complementary ; biosynthesis ; Gene Library ; Hepatitis B, Chronic ; complications ; genetics ; virology ; Humans ; Liver Cirrhosis ; genetics ; virology ; Polymerase Chain Reaction ; Recombination, Genetic ; Transcription, Genetic ; genetics

OBJECTIVETo evaluate the inhibition effects of DNAzymes specific to Hepatitis B Virus(HBV) s gene and e gene on the expressions of Hepatitis B surface antigen(HBsAg) and Hepatitis B e antigen(HBeAg).

METHODSDNAzymes DrzBS and DrzBC specific to HBV s gene ORF A157UG and e gene ORF A1816UG, respectively, were designed and synthesized. The inhibition effects of DrzBS or DrzBC on the expressions of HBV s and e genes were observed in 2.2.15 cells.

RESULTSThe expression of HBV s or e genes was dramatically depressed after 2.2.15 cells treated by DrzBS or DrzBC. The concentration for effective inhibition was within 0.1-2.5 micromol/L and the inhibition showed a dose dependence within that concentration range. The maximum inhibition was 94.2% and 91.8% for DrzBS and DrzBC, respectively. The inhibition was maintained for 72 hours. The efficiency of inhibiting HbsAg, HbeAg in 2.2.15 cells by DrzBS, DrzBC was higher than that by antisense oligonucleotides for the same target genes. The concentrations for effective inhibition of the DNAzymes were at least 10-fold lower compared with antisense oligonucleotides. Neither inhibition on the replication of HBV DNA nor toxicity to 2.2.15 cells was observed.

CONCLUSIONDrzBS and DrzBC can highly block the expressions of HBV s gene and e gene in 2.2.15 HBV cell model and are proved a specific and effective anti-HBV gene therapeutic means.

DNA, Catalytic ; pharmacology ; therapeutic use ; DNA, Viral ; analysis ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Genetic Therapy ; Hepatitis B ; therapy ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B e Antigens ; genetics