Bone Diseases ; complications ; pathology ; Bone Diseases, Infectious ; complications ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; complications ; microbiology ; Male ; Middle Aged ; Necrosis ; Osteomyelitis ; complications

Objective To construct the vector that expresses the fusion protein of HIV-Tat protein and red fluorescent protein(mCherry) in mammalian cells,and observe by fluorescence microscopy the intracellular transduction and localization of recombinant protein in cells,in order to obtain a useful tool for the study of the uptake mechanism and intracellular localization of HIV-TAT.Methods With the designed primer coding mCherry sequence,the mCherry gene was amplified by PCR with the vector pmCherry-C2 as template,and inserted into vector pET14b-His-TAT to construct the expression vector pET14b-His-TAT-mCherry.The constructed vector was then transformed into E.coli BL21(DE3),which had been identified by PCR and double digested with restriction endonuclease,followed by sequencing.After IPTG induction,the recombinant protein of His-TAT-mCherry was lyzed and analyzed with SDS-PAGE.Purified His-TAT-mCherry recombinant protein was added to Hela cells and the fluorescence was observed to evaluate the transduction efficiency.Results The results of identification by PCR,digestion with restriction endonuclease and sequencing indicated that the vector His-TAT-mCherry was correctly constructed.His-TAT-mCherry fusion protein was expressed in mammalian Hela cell line and purified successfully,and the fusion protein showed cellular transduction activity.It was found by fluorescence microscopy that the red fluorescence protein located mainly over the cytoplasm,and also the membrane to some extent.Conclusion The expression vector is successfully constructed for HIV-TAT labeled with mCherry sequence.Effective expression and purification of this fusion protein is achieved.It has been observed that the constructed vector may be expressed in mammalian Hela cell under active condition.Thus,it might be useful in the study of uptake mechanism and intracellular localization of HIV-TAT.

AlM: To compare the different formulas in intraocular lens(lOL) power calculations on high myopia after LASlK with cataract patients who received the phacoemulsification and intraocular lens implantation ( Phaco+lOL) .
METHODS:This was a retrospective study. Data from 102 eyes of 61 cataract patients with high myopia, who received Phaco+lOL after LASlK, were collected. Patients were divided into three groups by formulas in lOL power calculations before surgery: Holladay group ( 11 cases, 21 eyes), Haigis-L group (30 cases, 47 eyes) and SRK-T group (20 cases, 34 eyes). The ratios of equivalent lOL power after LASlK were predicted and the accuracy of their predictions were compared 3mo after surgery. lt was an index of reliability of the formulas outcomes.
RESULTS:The ratios of equivalent lOL power after LASlK using the Holladay, Haigis-L and SRK-T formulas were 0. 86 ± 0. 41D, 0. 43 ± 0. 30D and 1. 27 ± 0. 58D, respectively . There was no significant difference between Holladay group and SRK-T group ( t=-0. 271, P=0. 625>0. 05 ). However, significant difference was found between Haigis-L group and Holladay group ( t=2. 249, P=0.047<0. 05), Haigis-L group and SRK-T group (t=6. 012, P=0. 031<0. 05).
CONCLUSlON:The Haigis-L formula is more accurate than the SRK-T and Holladay formulas in predicting lOL power for cataract patients with high myopia after LASlK.

Objective To investigate the spousal transmission of human immunodeficiency virus (HIV)and its related factors in HIV epidemic area,which can be beneficial to prevent HIV from transmitting.Methods Three hundred and forty-six couples with one spouse were anti-HIV positive were cross-sectionally investigated.Blood samples were taken from the spouse of subjects whose anti- HIV were positive to detect the anti-HIV antibody and from 70 acquired immune deficiency syndrome (AIDS)patients to do the sequencing of the serum HIV provirus DNA.Results In 346 couples,99 were infected by spousal transmission and its transmission rate was 28.6%.One spouse of 125 couples were infected with HIV by paid blood donation,14.4%(18)of the other spouse were infec- ted by spousal transmission.One spouse of 135 couples were infected by paid blood transfusion, 23.7%(32)of the other spouse were infected by spousal transmission.Eighty-six couples were infec- ted by extramarital sexual contact,49(57.0%)got spousal transmission.Thirty-seven(69.8%) subjects were infected by husband-to-wife transmission and 12(36.4%)were from wife to husband. The difference between them was significant(P

BACKGROUND: When central nervous system is injured, re-expression of nestin protein may enhance the anti-injury ability of cells and be advantageous to the repair of focus of injury.OBJECTIVE: To explore the reaction of nerve stem cell (NSC) in permanent brain ischemia through NSC migration and the change of nestin protein expression.DESIGN: A randomized and controlled verification research with experimental animals as subjects.SETTING: Anatomy teaching and research offices in a training school and a university.MATERIALS: The experiment was done in the Teaching and Research Office of Humane Anatomy in Medical College of Xi'an Jiaotong University from October 1999 to January 2001. Totally 75 healthy SD rats were selected and randomly divided into normal control group, experiment group and sham-operation group. Twenty-five animals were in each group. Heads of animals were cut and brain was got out at the 1st, 3rd, 7th, 14th and 28thdays after operation, 5 animals at each time.METHODS: The model was rats with permanent cerebral ischemia. Immunohistochemical dyeing methods were used to observe NSC migration,change of marker of NSC and nestin protein at the 1st, 3rd, 7th, 14th and 28th day after cerebral ischemia.MAIN OUTCOME MEASURES: ①Results of immunohostochemicaldyeing. ②Migration length of nestin+ cells in anterior subentricular zone (SZa) region of brain tissue at normal status and at different time points after cerebral ischemia. ③Number variation of nestin+ cells at different timepoits after ischemia near the ischemic region.RESULTS: Through nestin immunohistochemical dyeing, it was found that NSC in normal brain tissue mainly existed in subependymal zone (SEZ)region. NSC of SEZ migrated in the direction of ischemic region along ventri- corpus callosum after ischemia. Among them, it reached the farthest at the 7th day after ischemia. More nestin+ cells appeared near ischemic region at the 1st day, and then reduced little by little 3 days later.CONCLUSION: NSC has certain reactive ability to ischemic brain injury.Expression of nestin protein near ischemic region may be a kind of protection to injury.

Objective To investigate the clinical value of blood vessel diameter tracking and X-strain technology in evaluating the carotid elasticity in patients with diabetes.Methods Thirty-eight patients who were confirmed as diabetes without complications were enrolled in this study as the patient group and thirtyseven healthy volunteers matched with the patient group with sex,ages were typed as controls.The parameters reflecting common carotid elasticity:pulse wave velocity ( PWV),compliance coefficient (CC),stiffness index ( β),endovascular circumferential strain,strain rate,strain time ( EN _ CS,EN _ CS_ T,EN_CSR,EN_CSR_T),adventitial circumferential strain,strain rate,strain time (EP_CS,EP_CS_T,EP_CSR,EP_ CSR_ T),radial strain ( RS),radial strain rate (RSR) and radial strain time (RST) were measured using blood vessel diameter tracking technique and X-strain technique.Significant difference between the two groups and correlations among these variables were evaluated.Results In patient group,PWV and β were significantly higher than those of the control group,while variables like CC,EN_CS,EN_CSR,EP_CS,EP_CSR,RS and RSR were lower with P ＜0.05.All strain time prolonged in patient group ( P ＜0.05).No significant differences were observed in longitudinal strain index.Furthermore,CC was inversely related with PWV( r =- 0.872,P ＜0.001 ),and age,systolic pressure,β were positively related with PWV ( r =0.322,P =0.005; r =0.384,P =0.001; r =0.927,P ＜0.001) in patients group.Conclusions The stiffness and compliance indexes measured by blood vessel diameter tracking technique and the circumferential and radial strain index obtained by X-strain technique can reflect vessel elasticity change of patient with diabetes objectively.

Objective To analyze the emergercy epidemiological characteristics of coeualties with head in-jury in Shanghai. Method By a prospective study,the data of 18 076 casualties with head injury during the whole year 2004 collected from 12 joint hospitals in Shanghai were documented well in the unified survey tables with wide-range items failed in by the attending physician, who got the firsthand information from patients and witness.The data were analyzed by using SPPS version 11.5 software. Results Of 18 076 casualties with head injury,the ratio of male to female was 2.11: 1.The mean age of female was older than that of male (t=10.575, P<0.01).The highest incidence of casualties occurred in people of twenties (24.7 % ). The local residents of Shanghai ac-counted for 34.2% of casualties. More casualties often occurred in December,January,Mareh and August than in he rest. Of 5.1% casualties with head injury were assochted with labour work.The leading cause of injury was dif-ferent in patient cohorts of different ages. Fall was the most main cause of trauma in children cohort (aged<14)and the senile patients cohort (aged > 60). The violert assault and traffic accident caused the most head injuries in the adolescent people cohort ( aged 15～34 years old) and the young people cohort ( aged 35～59 year old). The majority of casualties (85.5%) received CT scan.The scalp laceration (40.2% of patients) was seen more often than other types of injury . The mortality of easualties with traumatic brain injury was 0. 5 % . Conclusions The kmowledge of epidemiologieal aend of emergency deparhnent visitors with head injury is amportant guidance to physicians arranging emergency medical resources rationally and formulating a comprehesive prevention stategy of castahies with head injury.

C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Hernia, Inguinal ; immunology ; surgery ; Humans ; Infant ; Interleukin-6 ; blood ; Laparoscopy ; methods ; Tumor Necrosis Factor-alpha ; blood

Objective To construct and identify the recombinant retroviral vector containing five copies of hypoxia responsive elements (5HRE)and neurotrophin-3 (NT-3 ).Methods Using PCR,enzyme digestion and DNA ligase,5HRE and human derived NT-3 were cloned into the retroviral vector plasmid (pLNCX)to construct the recombinant retroviral vector plasmid pLNCX-5HRE-SV40-NT3-IRES-EGFP.The retrovirus RV-5HRE-NT3 was packaged in the PT67 cells,and then it was purified and concentrated by high-speed centrifugation.After infected for 48 h with the concentrated retrovirus,the number of the EGFP positive cells in the NIH 3T3 cells was counted by fluorescence activated cells and sorted to calculate the retrovirus titer.Results The retroviral vector plasmid,pLNCX-5HRE-SV40-NT3-IRES-EGFP,was successfully constructed,and the retrovirus was packaged and defined as RV-5HRE-NT3.After purification and concentration,the retrovirus titer reached 9.1 × 10 6 cfu/mL. Conclusion The recombinant retroviral vector which carried out hypoxia-regulated expression of NT-3 was successfully constructed.It may provide basis for studies on hypoxia-regulated expression of the exogenous genes.

ObjectiveTo investigate the application of rapid immunohistochemical staining technique in intraoperative frozen section diagnosis of thyroid neoplasm.Methods MaxVision one-step rapid immunohistochemical staining technique was used to detect the expression of CK19,HBME-1,and Gal-3 in frozen section of papillary thyroid carcinoma(PTC)andthyroid benign lesions.MaxVision conventional immunohistochemistry of frozen remaining tissue was served as control.ResultsMaxVision one-step rapid immunohistochemical staining technique could be completed in 20 minutes.The positive localizations of three markers detected by rapid immunohistochemistry were similar to conventional immunohistochemistry, in general.The expression of CK19 was located in cytoplasm and cellular membrane.Gal-3 and HBME-1 were mainly detected in follicular luminal border and/or surface of papilla. The staining intensity in rapid immunohistochemistry was stronger than that in conventional immunohistochemistry. The positive rates of CK19,HBME-1,and Gal-3 by rapid immunohistochemistry in frozen sections were: 0 (0/28),10.7 ％ (3/28),0 (0/28),respectively,for benign lesions (nodular goiter,Hashimoto thyroiditis,thyroid adenoma); and 94.9 ％(37/39),92.3 ％ (36/39),92.3 ％ (36/39),respectively,for PTC.The expression of three markers between thyroid benign lesions and PTC had a significant difference (x2 =59.326,55.861,44.605,all P ＜ 0.001).In benign lesions,the rate of same case with two and more positive markers was 0,while in PTC it was 100 ％ and significantly different (x2 =67.000,P ＜ 0.05).ConclusionMaxVision one-step rapid immunohistochemical staining technique could be applied in intraoperative frozen section diagnosis.Detecting CK19,HBME-1,and Gal-3 expression in intraoperative frozen section has an auxiliary value for diagnosis of PTC.