0.1).One(1.3%) of them suffered from intracranial hemorrhage.Conclusions 1.(L-ASP) may affect the function of coagulation system,such as prolonged APTT and hypofibrinogenemia.2.There was no statistical difference in side effects of coagulation,in use of domestic L-ASP and foreign L-ASP,intravenous dosing and intramuscular dosing.

Objective To explore the risk factors of elderly people with pulmonary TB,and to provide the scientific evidence of prevention and interventions for tuberculosis among the old people in Nanchang City.Methods 1∶1 case control study was performed .A total of 390 pulmonary TB patients over 60 years old were selected as case group.One healthy person with the same gender and within 2 year age difference was selected to match each case.Interviews were carried out with a uniformly designed questionnaire.Logistic regression models were used for analysis.Results A total of 120 smear positive tuberculosis patients and 270 smear negative tuberculosis patients were investigated.And 72.82% male and 27.18% female of the 390 TB patients were investigated.Average age of patients was 70.34 ±7.75.Multivariate condition logistic regression analysis showed smoking(OR =2.359,95%CI:1.368 -4.068),contacting tuberculosis(OR =3.357,95%CI:1.854 -6.075),BMI 18.5 -24.9(OR =0.175,95%CI:0.056 -0.546),education (OR =0.110,95%CI:0.036 -0.332),annual average income (OR =0.475,95%CI:0.332 -0.681),per capita living space(OR =0.946,95%CI:0.920 -0.973)and drinking tea (OR =0.398,95%CI:0.268 -0.592)were the influencing factors(P <0.05).Conclusion Health education should be promoted auording to the risk factors,and patients manage ment should be streng thened.

Objective To evaluate the surgical approaches and therapeutic effect of lymphatic malformations located in head and neck in children. Methods Eleven cases of lymphatic malformations in the region of head and neck in children encountered between Jan. 1998 and Dec. 2008 in Peking University First Hospital were retrospectively analyzed. Initial diagnosis was made based on the physical examination and then confirmed by MR and Enhanced CT imaging. Surgical therapy was used for patients with lymphatic malformation which exceeds 4 cm. The operative technique was as follows: mass resection and superficial parotidectomy (4 cases), mass resection and total parotidectomy (2 cases), mass resection with neck dissection (2 cases), mass resection with neck dissection and sternotomy (1 case), marginal mandibular branch of facial nerve dissection and mass resection (2 cases). Dissection outside the false capsule was applied during the operation and facial nerve was dissected from bole to terminal arborization. Results The mass was completely removed in all 11 cases without organ dysfunction and obvious dysfigurement. The cure rate was 100%. Three cases suffered from a branch of facial nerve paralysis because of tension and 1 case had a Homer's syndrome after operation. One ease needed a blood transfusion (150 ml ) during the operation. All cases have been followed up with excellent results from 6 to 121 months, 32 months of the median, no mass recurrence. Conclusions Dissection outside the false capsule of mass and dissection of facial nerve were applied in the surgical treatment of huge lymphatic malformations. These methods are effective in the preservation of function and avoidance of abnormality.

OBJECTIVETo observe the differentiation and distribution of epidermal stem cell (ESC) after skin soft tissue expansion, and to initially probe into the growth mechanism of expanded skin tissue.

METHODSSamples of normal skin and expanded skin (mean effusion period 45 days) were harvested from head and cervical region in 15 patients who underwent II stage surgery after skin expansion. Samples were divided into scalp adjacent to the center of expander group (expanded scalp, 3 cm from the vertical axis of the expander), scalp from lateral part of the expander group (expanded scalp, 5 - 7 cm lateral to the vertical axis of the expander), cervical skin expansion group, un-expanded scalp control group, and un-expanded cervical skin control group, according to the position of skin harvested. The tissue structure of skin in each group was observed with HE staining, and the differentiation and distribution characteristics of cytokeratin 19 (CK19) positive cells were observed with immunohistochemical staining.

RESULTSCompared with those in the un-expanded control groups, uneven, relatively thickened and obviously folded epidermis with more cell layers and cells with obvious aggregation close to the basal layer were observed in the expanded groups, but those cells were not well-arranged and the transition of polarity was not obvious. The continuity of CK19 positive cells in the basal layer of skin was observed in each of the expanded group with immunohistochemical staining, and positive cells increased obviously and arranged in multilayer in certain parts of basal layer. Clustered or dispersed CK19 positive cells were also observed outside the basal layer. No above-mentioned phenomenon was observed in the un-expanded control group.

CONCLUSIONSThe proliferation and differentiation of ESC with ectopic distribution may enhance the repair process after skin soft tissue expansion.

Cell Proliferation ; Dermis ; cytology ; Epidermis ; cytology ; Humans ; Stem Cells ; cytology ; Tissue Expansion ; Wound Healing

A simple, reliable and sensitive liquid chromatography-isotope dilution mass spectrometry (LC-ID/MS) was developed and validated for quantification of olanzapine in human plasma. Plasma samples (50 microL) were extracted with tert-butyl methyl ether and isotope-labeled internal standard (olanzapine-D3) was used. The chromatographic separation was performed on XBridge Shield RP 18 (100 mm x 2.1 mm, 3.5 microm, Waters). An isocratic program was used at a flow rate of 0.4 m x min(-1) with mobile phase consisting of acetonitrile and ammonium buffer (pH 8). The protonated ions of analytes were detected in positive ionization by multiple reactions monitoring (MRM) mode. The plasma method, with a lower limit of quantification (LLOQ) of 0.1 ng x mL(-1), demonstrated good linearity over a range of 0.1 - 30 ng x mL(-1) of olanzapine. Specificity, linearity, accuracy, precision, recovery, matrix effect and stability were evaluated during method validation. The validated method was successfully applied to analyzing human plasma samples in bioavailability study.
Antipsychotic Agents ; blood ; pharmacokinetics ; Benzodiazepines ; blood ; pharmacokinetics ; Biological Availability ; Chromatography, Liquid ; methods ; Humans ; Indicator Dilution Techniques ; Isotope Labeling ; Reproducibility of Results ; Sensitivity and Specificity ; Tandem Mass Spectrometry ; methods

Objective To study the Th17/Treg (regulatory T cells) immunoregulation in patients coinfected with TB and HIV before and after HAART( highly active anti-retroviral therapy).Methods 10 HIV cases coinfected with TB ( HIV/TB group) and 10 cases infected with HIV only ( HIV group) received HAART.PBMCs were stained and immunophenotyping of Th17( IL-17 expressing T cells) and CD4 + CD25 +T cells (Treg) were analysed by flow cytometry.Results The pre-treatment patients tended to have lower Th17 cells and higher Tregs cells compared to post-treatment( 1.90％ ± 0.9％ vs.4.65％ ± 1.48％,16.48％ ±4.91％ vs.8.29％ ± 3.13％ respectively).The percentage of IL-17 before and after HAART were 1.90 ± 0.9％ vs.4.65 ± 1.48％ respectively in HIV/TB group patients ( P ＜ 0.01 ).The difference between the percentage of IL-17 before and after HAART in the HIV/TB group and the HIV group were 2.65 ± 1.62％ vs.0.67％ ± 0.46％ respectively (P ＜0.01 ).IL-17 expressing T cells were increased faster after HAART in the former group than the latter.The percentage of Treg before and after HAART were 16.48％ ±4.91％ vs.8.29％ ± 3.13％ respectively in HIV/TB group ( P ＜ 0.01 ).The difference between the percentage of Treg before and after HAART in the HIV/TB group and the HIV group were 8.91％ ±4.82％ vs.2.63％ ± 2.34％ respectively ( P ＜ 0.01 ).Treg were decreased more rapidly after HAART in the former than the latter.Conclusions TB and HAART both had an effect on the Th17/Treg ratio of HIV/ TB co-infected patients,which can cause increased Th17 expression,the later plays a pro-inflammatory role.TB and HAART can decrease Treg expression and enhance anti-inflammation response.The fact that Th17/Treg disorder are more likely to exist in patients with HIV/TB co-infection after HAART for one month suggests a potential role for Th17/Treg imbalance leading to tuberculosis-associated immune reconstitution inflammatory syndrome during patients receiving HAART period.

Objective To observe how complement regulatory protein CD59 expression on T cells in HIV infected patients and discuss the meaning of how highly active antiretroviral therapy(HARRT) affect CD59 expression. Methods 48 HIV infected patients and 14 healthy donors were performed in this study.Patients were divided into Naive group and On-HARRT group according to HARRT or not. The peripheral blood samples were collected and cell surface cytokines were stained, and then were evaluated with the BD FACS Canto flow cytometry, after that, the expression of CD59 on CD4+T, CD8+T and memory CD45ROCD4+T cells were analyzed, and HIVRNA were detected with PCR, then compare the results between groups. Results Compared with healthy donor, the expression of CD59 on T cells in HIV infected patients are significantly higher(P＜0.05), most of that expressed on CD45ROCD4+T cells(P＜0.05).Compared with Naive group, the CD59 expression on CD4+T cells in On-HARRT group decreased significantly but it is still higher while compared with healthy control(P＜0.05). CD59 expression on CD4+T cells is correlated with HIVRNA and CD4+T cells count(R2=0.2181, P=0.0247; R2=0.1586, P=0.0486). Conclusion HIV infection can cause CD59 expression increase on CD4+T cells and HARRT can decrease its expression. This increase may be related to HIV immune escape and CD4 +T cell function inhibition, and HARRT can partially reverse this immune disorder.

OBJECTIVETo observe the distribution of epidermal stem cell (ESC) after soft tissue expansion, and to explore dynamic change in ESC under mechanical stress and kinetic mechanism of skin expansion.

METHODSSkin samples were collected from patients after expansion of the scalp. They were divided into three groups: A group (scalp harvested 3 cm away from the center of dilator), B group (scalp tissues at the edge of dilator), and control group (scalp without dilatation). The tissue structures were observed with optical microscope with HE staining. The distribution and differentiation characteristics of cell keratin 19 (CK19) positive cells were observed with inverted phase contrast microscope after immunohistochemistry staining.

RESULTSHE staining showed that the epidermis was thickened and distributed densely with uneven, rugged and increased layers in A, B groups. With immunohistochemistry staining, CK19 positive cells appeared in multilayers in basal membrane, a few of them were in cluster or dispersed , with" hollowing" structure formation. These phenomena were not seen in control group.

CONCLUSIONESC can proliferate with abnormal distribution and "hollowing" structure formation after mechanical dilatation, which may be related to dynamic changes in basal layer cells.

Adolescent ; Adult ; Cell Proliferation ; Cellular Structures ; Epithelial Cells ; cytology ; Humans ; Keratin-19 ; metabolism ; Male ; Stem Cells ; cytology ; Stress, Mechanical ; Tissue Expansion ; Young Adult

OBJECTIVETo evaluate the diagnostic applicability of human papillomavirus (HPV) liquid chip assay which is based on Luminex XMAP System, and perform a HPV epidemiologic study with the liquid chip in women of Shandong province.

METHODSTo detect HPV genotypes on a 96-well plate with the liquid chip which can simultaneously detect and identify 26 common HPV genotypes in a total of 2925 cervical scrapes obtained from gynecological outpatients as well as to analyze the relationship between HPV types and different cervical diseases by studying the distribution of HPV genotypes and pathologic diagnosis.

RESULTSAmong 639 cases who performed pathologic/cytological and histological diagnoses, 184 cases are in group of normal cytology, 266 cases in group of, 77 cases in group of cervical intra-epithelial neoplasia (CIN) I, 7 cases in group of CIN I - II, 46 cases in group of CIN I - II, 46 cases in group of CIN I - II and 13 cases in group of cervical cancer. The overall incidence of HPV in our samples is 36.0% (1054/2925) and 23 types of all 26 types on liquid chip are found. The most common genotypes found are HPV-16 (26.75%), HPV-52 (25.75%), HPV-58 (10.47%), HPV-18 (8.87%) and HPV-11 (6.94%). Among all the positive types, 87.32% are high-risk HPV and 13.68% are low-risk HPV genotypes. Both single and multiple types are easily identified, showing 66.22% ( n = 698) single type and 33.78% ( n = 356) multiple types. Of all the 1054 HPV-positive cases, 261 (24.8%) is occupied by women 21 to 25 years of age and progressively lower by older age groups, reaching 4.9% by women between 51 to 67 years old. The incidence of HPV in our samples is 23.37%, 33.08%, 54.54%, 57.14%, 82.61%, 91.30% and 100% for normal cytology, inflammation,CIN I ,CIN I - II, CIN II ,CIN III, and carcinomas specimens, respectively. Infections with more that one virus are common, accounted for 4.89%, 7.14%, 18.18%, 28.57%, 41.30%, 43.37% and 38.46% for normal cytology, inflammation, CIN I, CIN I - II, CIN II, CIN III, and carcinomas specimens, respectively. Based on the criteria of histology and pathology, the sensitivity, specificity, positive-predictive value and negative-predictive value of HPV liquid chip assay for detecting all cases of CIN II, III are 88.57%, 76.63%, 68.89% and 92.16% respectively. Conclusion The common types of HPV infection are 16, 52, 58, 18, 11, 6, 56 and 31. The HPV-positive rate increased along with the increase of grading on cervical lesions. There are more younger women among all the HPV-positive ones. Multiplex HPV genotyping by liquid chip appears to be highly suitable for diagnostic screening as well as the conduction of large-scale epidemiological studies.

Adolescent ; Adult ; Aged ; Cervical Intraepithelial Neoplasia ; epidemiology ; virology ; China ; epidemiology ; Female ; Gammapapillomavirus ; classification ; genetics ; Genotype ; Human papillomavirus 11 ; classification ; genetics ; Human papillomavirus 16 ; classification ; genetics ; Human papillomavirus 18 ; classification ; genetics ; Human papillomavirus 6 ; classification ; genetics ; Humans ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; epidemiology ; virology ; Uterine Cervical Neoplasms ; epidemiology ; virology ; Young Adult

OBJECTIVETo immunopurify human endometrial endothelial cells (HEEC) from fresh surgical specimens of endometrial cancers and normal endometrial tissues, and investigate their biological characteristics.

METHODSEndothelial cells of endometrial cancers and normal endometrial tissues were isolated using anti-CD31 conjugated magnetic microbeads. The isolated endothelial cells were cultured in vitro and their origins were identified. Their angiogenic characteristics were observed by MTT, wound healing, Transwell cell invasion and tube formation assays.

RESULTSFlow cytometry revealed that the immunopurification technique yielded endothelial cell purity of > 95% in all samples. All purified HEEC were characterized as endothelial cells on the basis of expression of the classical endothelial markers vWF and CD31 as shown by immunofluorescence examination. Although the tumor-associated HEEC didn't show more rapid proliferation than normal HEEC, they exhibited enhanced migration ability (P = 0.006), potent invasiveness (P = 0.033), and elevated tube formation in vitro (P = 0.029).

CONCLUSIONSHuman endometrial endothelial cells can be efficiently isolated from endometrial cancer and normal endometrial tissues by immunomagnetic methods. Tumor-associated HEEC exhibit enhanced migratory ability, potent invasiveness, and elevated tube formation in vitro.

Adult ; Aged ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; cytology ; metabolism ; pathology ; Endothelial Cells ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism