Objective To establish the methods of isolating and culturing human ovarian endometriosis-derived microvascular endothelial cells (OEMEC).Methods The tissues of human endometriotic cyst of ovary were finely minced with scissors,then digested by collagenase Ⅰ,Ⅱ and trypsinethylene diamine tetraacetic acid (EDTA).The cells were purified by using centrifugation of 2000 r/min speed.OEMEC were identified by light microscope and transmission electron microscope observing CD34,FⅧ-Rag and Weibel-Palade in microvascular endothelial cells.Results The OEMEC grew as confluent monolayer like cobblestones under light microscope.CD34 and FⅧ-Rag were expressed strongly,and the percentages of CD34 and FⅧ-Rag positive cells were 91.4％ and 92.5％.Weibel-Palade bodies could be observed under transmission electron microscope.The time of cell doubling proliferation was 4.5 days.Conclusion The established system of isolating OEMEC would provide lab base for studying the mechanisms of angiogenesis in endometriosis lesions.

Objective To conduct an in vitro study of the growth of Pseudomonas aeruginosa,Staphylococcus aureus and Acin- etobacter spp.,and evaluate their response to various concentrations of tumor necrosis factor(TNF)-?,interleukin (IL)-1?, and IL-6.Methods To monitor the growth of bacteria incubated with the cytokines TNF?,IL-1?and IL-6 that were added to RPMI 1640 medium in various concentrations (10,50.100,500 pg,1 and 10 ng) at 2,4 to 6,8 and 16-18 h.The bacterial concentration was estimated when the mixtures of cytokines and specific neutralizing monoclonal antibodies (MoAbs) were in- cubated.Results We found that all three bacterial species showed concentration-dependent growth enhancement when incubated with one or more tested cytokines.Blockade by specific neutralizing cytokine significantly inhibited cytokine-induced growth. When compared with control,the 6 h growth response was maximal with IL-1?for Staphylococcus aureus and Acinetobacter spp.,and with IL-6 for Pseudomonas aeruginosa.Conclusions In this study we provide additional evidence for a newly de- scribed mechanism for bacterial proliferation in the presence of exaggerated and protracted inflammation.The effect that cyto kine-induced growth enhancement inhibited by specific neutralizing cytokine MoAbs may be useful for antimicrobial therapy.

Objective To prepare,purify and validate specific monoclonal antibodies(McAb) against fragment in C-terminal telopeptides of collagen type Ⅱ(EKGPDP)for clinical diagnostics and related research of osteoarthritis(OA).Methods 8 BALB/c mice were immunized with EKGPDP-KLH antigen complex.McAb against EKGPDP fragements were prepared by hybridoma technique.Immunoglobulin classes and subclasses were determined using an Immuno-Type~(TM)mouse McAb isotyping kit.Ascites were producted and McAb were purified by saturation ammonium sulfate(SAS)precipitation and protein G chromatography.Specificity and immunoactivity of antibodies were detected by indirect enzyme-linked imrnunosorbant assay(ELISA).Cartilage specimens were analyzed by immunohistochemical method.Results The hybridoma cells were obtained.10 IgG and 3 IgM single colonies were picked out by limiting dilution and ELISA kit.The titers of McAb in ascites were from 2.8?10~4 to 5.1?10~5.The McAb showed the characteristics of no cross reactions with KLH,BSA,cell culture supernatant,type Ⅰ,Ⅲ collagens and whole type Ⅱ collagen.The method of SAS could get better recovery,immunoactivity of the McAb than protein G chromatograply(t=25.26;P

OBJECTIVESTo investigate whether antisense human telomerase reverse transcriptase (hTERT) could inhibit the activity of telomerase and the proliferation of K562 cells.

METHODSThe antisense plasmid was constructed by reverse insertion of hTERT PCR product into plasmid pLNCX-neo. Then the constructed plasmid was introduced into K562 cells by liposomes-mediated DNA transfection. The inhibition effects of telomerase on the proliferation of K562 cells were analyzed by MTT and colony formation assay, the telomerase activity of K562 cells by TRAP-PCR ELISA methods.

RESULTSThe growth rate of antisense hTERT transfected K562 cells was significantly lower than those of the controls, and the colony formation capacity of the transfected cells decreased significantly (P < 0.01), the colony number is (100.33 +/- 7.57)/10(3) cells, (92.67 +/- 5.86)/10(3) cells and (50.33 +/- 6.11)/10(3) cells for control K562 cells, K562 neo cells and antisense hTERT transfected HL60 cells, respectively. The telomerase activity of antisense hTERT transfected K562 cells was significantly inhibited.

CONCLUSIONThe expression of an antisense sequence to the mRNA sequence of telomerase protein subunit can inhibit the activity of telomerase, slow the cell growth and inhibit the capacity of colony formation of K562 cells.

Cell Division ; drug effects ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Telomerase ; drug effects ; genetics ; metabolism ; Transfection

Viral hepatitis was the most common infectious disease in china. But the diagnosis and treatment were varied because the viral hepatitis patients were hospitalized in different kinds of hospital such as infectious disease hospital, general hospital and Chinese medical hospital. It was necessary to know clinical characters and information of viral hepatitis patients in different hospitals. The general information, subtype distribution, prognosis, complication, medication and relations of onset with solar term from 41 180 viral hepatitis patients based on HIS data were analyzed. It was found that the age of patients between 18 to 59 years old was most; most patients were males. The national basic medical insurance was the most type of payment. The outcome of viral hepatitis in the youth and female were better than that in the old and male. Acute hepatitis was easer to restore than chronic hepatitis. Liver cirrhosis and hepatocellular carcinoma were the two most complications. The peak of onset was during summer solstice, slight heat and great heat. The most common Chinese medicine was Diammonium glycyrrhizinate and the most common western medicine was reduced glutathione. The combination of D. glycyrrhizinate with reduced glutathione, polyene phosphatidylcholine and thymosin was the main pattern. But It was not knew if the combination of western and Chinese medicine was the most effective therapy to protect liver function. It was necessary to take deeply research of the relationship between the combination therapy and their effectiveness.
Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Glutathione ; therapeutic use ; Glycyrrhizic Acid ; therapeutic use ; Hepatitis, Viral, Human ; drug therapy ; Hospitals ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVE: To establish an inductively coupled plasma-mass spectrometry (ICP-MS) method for determination of 24 elements in human hair. METHODS: The samples were digested with microwave digestion instrument. ICP-MS was applied to determine 24 elements in human hair using indium (115In) as an internal standard. The established method was applied to determine element concentration in normal group (56 samples) and heroin abuse group (10 samples). RESULTS: The limits of detection ranged from 0.000 3 microg/g to 10.14 microg/g. Measured value of the standard materials were basically consistent with the standard value. The contents of magnesium, gallium and barium in hair of heroin addicts decreased after rehabilitation treatment. CONCLUSION The method is sensitive and accurate for determination of 24 elements in human hair.
Adolescent ; Adult ; Aged ; Barium/analysis* ; Child ; Child, Preschool ; Female ; Gallium/analysis* ; Hair/chemistry* ; Heroin Dependence/rehabilitation* ; Humans ; Magnesium/analysis* ; Male ; Mass Spectrometry/methods* ; Metals, Heavy/analysis* ; Microwaves ; Middle Aged ; Sensitivity and Specificity ; Spectrophotometry, Atomic ; Trace Elements/analysis* ; Young Adult

The myelodysplastic syndromes (MDS) include a diverse groups of clonal and potentially malignant bone marrow disorders. Evidences exist that microenvironment cells from MDS marrow show functional abnormalities, which may be relevant to the incidence of such a disease. Mesenchymal stem cells (MSCs) are a very important component of hematopoietic microenvironment. This study was supposed to investigate the biological characteristics and functions of MSC derived from patients with MDS in low-risk. MSCs from bone marrow samples of 11 low-risk MDS patients were isolated, cultured and expanded. Morphology, immunophenotype and osteoblasts differentiation were analyzed. Their capacity of proliferation and hematopoietic supporting in vitro were measured. A real-time quantitative reverse transcriptase polymerase chain reaction method (RQ RT-PCR) was used for detecting the expression levels of relative cytokines and chemokines in MSC. MSCs from healthy donors were used as controls. The results showed that the culture-expanded cells from MDS patients displayed a typical fibroblast-like morphology. Cells were positive for SH2 (CD105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. After induction, these cells could differentiate into osteoblasts. The proliferative ability of MSCs in MDS patients were not different from those of MSC isolated from normal bone marrow (p > 0.05), however, their capacity of hematopoietic supporting in vitro were significantly weaker (p < 0.05). RQ RT-PCR detection indicated that the SDF-1 gene expression level in MSCs of low-risk MDS patients was significantly higher than that in MSC derived from healthy donors (p < 0.01). It is concluded that the abnormal function of MSC influences the regulation of hemotopoiesis in the bone marrow microenvironment of MDS patients. It is worthy to further investigate the new clue in etiological mechanism and therapeutic strategies for MDS.
Bone Marrow Cells ; pathology ; Chemokine CXCL12 ; genetics ; metabolism ; Hematopoiesis ; Humans ; Mesenchymal Stromal Cells ; pathology ; physiology ; Myelodysplastic Syndromes ; pathology ; physiopathology ; Risk Factors

Verapamil is used in the treatment of hypertension, angina pectoris, and atrial fibrillation. Recently, several studies have demonstrated that verapamil increased the optic nerve head blood flow and improved the retrobulbar circulation. All these show that verapamil is potentially useful for ophthalmic treatment. Thus, the aim of this study is to investigate whether verapamil could protect human lens epithelial cell (HLEC) from oxidative stress induced by H2O2 and the cellular mechanism underlying this protective function. The viability of HLEC was determined by the MTT assay and apoptotic cell death was analyzed by Hoechst 33258 staining. Moreover, Caspase-3 expression was detected by immunocytochemistry and flow cytometry analysis. We also detected Caspase-3 mRNA expression by reverse-transcription-polymerase chain reaction and the GSH content in cell culture. The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil. Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content. Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by H2O2.
Angina Pectoris ; Apoptosis* ; Atrial Fibrillation ; Bisbenzimidazole ; Caspase 3 ; Cell Culture Techniques ; Cell Death ; Epithelial Cells* ; Flow Cytometry ; Glutathione ; Humans ; Hypertension ; Immunohistochemistry ; Optic Disk ; Oxidative Stress ; RNA, Messenger ; Verapamil*

Objective To explore the normal range of the insula volume of Chinese adults of the Han nationality and its relationship with age, which provide morphological data for the construction of database for Chinese Standard Brain.Methods This is a clinical multi-center study.One thousand Chinese healthy volunteers (age range = 18 to 70) recruited from 16 hospitals were divided into 5 groups, i.e.,Group A ( age range = 18 to 30), B ( age range = 31 to 40 ), C ( age range = 41 to 50 ), D ( age range =51 to 60), and E (age range =61 to 70).Each group contained 100 males and 100 females.All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.After three dimension data reconstruction, the volumes of bilateral insula were manually measured.The volume of bilateral insula were compared by paired sample t test.The insula volumes were compared between male and female by independent sample t test, and the differences among 5 age groups were compared by one-way ANNOVA.The relationship between the volumes of insula and age,sex or cerebral volume were analyzed using bivariate correlation, respectively.Results The left snd right side volume of insula before standarized were (7764±1165) and (7387±1128) mm3 respectively, after standarized were (8413±1201 ) and (7871±1140) mm3 respectively.The left insula volume were significant larger than that of right(t = - 10.565, - 16.014,P <0.01 ).The left and fight volume of insula were (8146±1181 ) and (7735±1113) mm3 for male, and (7393±1022) mm3 and (7050±1038) mm3for female.The left and right insula volumes for male were larger than the female's(t = 10.934,9.945 ,P <0.01 ).The left and right insula volume of male after standarized were (8779±1230 ) and (8224±1081 )mm3, female were (8043±1054) and (7515±1091 ) mm3 ,the left and right insula volume of male were larger than the female's ( t = 4.858,4.632, P < 0.01 ).The left insula volumes among Group A, B, C, D,E before standarized were ( 8268±1221 ), ( 8067±1107 ), ( 7869±1109), ( 7603±1111 ), ( 6997±934 )mm3 respectively,the right were (8028±1156), (7636±1075), (7294±986), (7249±1068), (6717±916) mm3 respectively, there were significant differences among 5 groups between left and right insula volume(F= -0.361,-0.337,P <0.01 ).The left insula volume of A,B,C,D,E after standarized were ( 9093±1105 ), ( 8679±965 ), ( 8810±1136), ( 8202±980), ( 7273±940 )mm3, the right were ( 8694±1005), (8136±1100), (8034±910), (7496±990), (6989±932) mm3, there were significant differences among 5 groups between left and right insula volume(F = -0.490, -0.512,P < 0.01 ).There was significant negative correlation between the volume and age ( before standarizded:r = -0.361, -0.337, after standardized r = -0.490, -0.512, P<0.05).Before and after standardized, there was significant correlation between the volume of right and left insula and cerebral volume ( r = 0.470,0.459, P < 0.05 ).Before and after standardized, there was significant correlation between the volume of right and left insula and weight( r = 0.141, 0.092, P < 0.05 ).Conclusions 1.5 T MR scanner has high resolution, for distinguishing the white matter from the gray matter, and provide morphological data of insula for the construction of database for Chinese Standard Brain.

OBJECTIVETo design and manufacture a reliable spine phantom used in the cross calibration and quality control of dual X-ray absorptiometry (DXA).

METHODSA hydroxyapatite quality control phantom was designed and made through three steps: solid water, bone phantom material and integration, then evaluated the phantom on four different types of DXA machines made by LUNAR company.

RESULTSAmong the four DXA densitometers, the Expert fan beam densitometer had the biggest accuracy errors of bone mineral density (BMD), bone mineral content(BMC) and area values while the other three one narrow fan beam (Prodigy) and two pencil beam densitometers-had small errors. Of the three indexes measured by all the machines, BMD error was the smallest (-15.4%-11.5%), with the Prodigy's BMD was most outstanding. BMD errors at the higher density ends were small, tend to be positive values while the errors at the lower ends were big, tend to be negative. In cross calibration, giving consideration to the differences between the both ends, it is better to use the regression equation to correct. The base line of precision error derived by scanning the phantom once a day for consecutive 25 days was better than that derived by scanning the phantom 25 times consecutively on the same day. As to precision error, the coefficient of variation (CV) of scanning-25 times-a day was the smallest (0.0043) while the CV of 12 adults measurements was the biggest (0.0078).

CONCLUSIONSThis phantom can be used in the quality control and cross-calibration of different types of DXA machines.

Absorptiometry, Photon ; Adolescent ; Adult ; Biocompatible Materials ; Bone Density ; Calibration ; Durapatite ; Female ; Humans ; Male ; Middle Aged ; Models, Anatomic ; Spine