Objective To probe into the diagnostic accuracy of endometrial sampling device in the detection of endometrial lesions by cytology. Methods Endometrial cytological examination was performed on totally 193 out-patients from January to May 2008,the results were compared with the histological results by diagnostic curettage (D&C). Results The sampling qualified rate of endometrial sampling device was 93.8% (181/193) while that of D&C was 99.0% (191/193,?2=7.412,P=0.006). Thirteen patients were diagnosed with endometrial malignant lesions by D&C. In these patients,the sensitivity and specificity of cytology was 69.2% (9/13) and 100.0% (166/166),respectively;and its accuracy rate was 97.8% (175/179). By D&C 18 patients were diagnosed with endometrial hyperplasia,in whom the sensitivity and specificity of cytology was 27.8% (5/18) and 98.8% (159/161),respectively,and the accuracy rate was 91.6% (164/179). Ten patients were confirmed as having endometrial benign lesions without hyperplasia by D&C,for whom the sensitivity and specificity of cytology was 60.0% (6/10) and 98.8% (167/169) respectively,and the accuracy rate was 96.6% (173/179). The accuracy rate of endometrial cytological examination for 80 infertility patients with satisfactory sample was 100%,when the main objective was to determine whether the endometrial gland secreted or not. Conclusion Cytological examination by endometrial sampling device is feasible in the detection of endometrial malignant lesions and inspection of endometrium for infertility patients,but further discussion is necessary in the detection of endometrial benign lesions.

Objective To explore the survival condition of hemodialysis patients in Ningxia Hui Autonomous Region. Methods Interviewed 15 patients with hemodialysis by applying Modified Grounded Theory Approach put forward by KinoSitaYasuHito,a Japanese scholar,the data of which were from qualitative inductive comparative analysis. Results As for the survival condition of hemodialysis patients in Ningxia,two relative themes were extracted, Helpless dialysis life Desire to return to society, the former contained 2 subtopics of hardship in life and susceptible mood, the latter contained 2 subtopics of cherish life and desire for care. Conclusion We should set up a social supporting system to improve the living condition of hemodialysis patients with the purpose of helping them return to the society.

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; Chromatography, Ion Exchange ; Humans ; Isoelectric Focusing ; Mass Spectrometry ; Peptide Mapping ; Peptides ; Sequence Analysis, Protein ; methods

BACKGROUND: There are many methods for investigating the physical development and the somatotype of child. Among them, Heath-Carter so matotype method is a comprehensive evaluation method on somatotype. In this method, 10 items of anthropometric indicators are selected from several dozens of items as assessment indicators for development and somatotype.OBJECTIVE: To analyze the rules and characteristics of soma totype development of Hah children in rural area of Huludao city, so as to supplement the essential data for physical anthropology.DESIGN: Normal children were chosen as the subjects, and cross-sectional investigation was carried out.SETTING: Department of Anatomy, Jinzhou Medical College. PARTICIPANTS: With cluster sampling, the Han children between 7 to 10 years old were selected from central primary school of Mingshuixiang Village of Suizhong County of Huludao city from July 2001 to September 2003. All the selected children should be verified healthy by physical examination at school. The children were divided into two groups according to gender, and each group was divided into 4 subgroups according to age,with 46 to 63 members in each group. There were 8 groups altogether.Complete data of 408 children were collected (male 213, female 195).METHODS: Each indicator was measured twice with Heath-Carter soma to type method and selected the average value. Specialists were responsible for the 10 indicators, and a database was established in computer according to age and gender. Calculated the following indicators in turn: endomorphic factor, mesomorphic factor and ectomorphic factor, coordinate values of X and Y on somatotype chart, Height/Weight1/3, mean of dimensional distances from the average somatotype to all somatotypes in the sample,difference between the two somatotypes in three-dimensional space, percentage of body fat and frequency distribution of each somatotype.MAIN OUTCOME MEASURES: Height, body mass, upper arm circum ference, calf circumference, intracondylar diameters of humerus and femur,skinfold of brachial triceps, subscapular skinfold, skinfold of anterosuperior iliac spine, skinfold of gastrocnemius muscle.RESULTS: All the 408 children entered results analysis. ①Height and body mass of 7 to 10 year old children increased as age went up, indicators of Height/Weight1/3 of 7,8,10 year old girls were higher than boys, that of 9 years old boys was higber than girls, which were related to that the growth of height of 8 & 9 year old boys (5.61 cm) was more than girls (3.88 cm).The content of body fat of girls was more than boys. The 3 somatotype values were 3.2-3.9, 2.3-2.6 and 3.0-3.4 in boys and 3.5-4.6, 1.9-2.6 and 3.3-3.5 in girls, so endomorphic factor and ectomorphic factor were the main factors both in girls and boys, and endomorphic gene in girls was higher than boys, which indicated that the content of body fat of rural children in Hulutao city was relatively high, their bodies were finely high and thin, their skeletal muscle was not so muscular and the hypodermic fat in girls was more abundant. Seen from the comparison of somatotype between boys and girls in each age group, the differences between two somatotypes in the three dimensional space of 7-, 8-, 9-, 10-11 year old groups were 0.46, 0.68, 1.03 and 0.61 respectively, except 7-year old group, there were significant differences in somatotype of boys and girls between each age groups (P ＜ 0.01). ②The average value of somatotype of Han boys in rural area was 3.4-2.5-3.2, which belonged to medial type; that of girls was 4.0-2.3-3.4, which beglonged to ectomorphic endomorphic type. Endomorphic factor and ectomorphic factor were the dominant factors both in boys and girls, whereas mesomorphic factor was relatively low; the frequency of somatotype kept changing as the increase of age. ③Compared with the data at home and abroad, the endomorphic factor of the Han children in rural area in Huludao City was relatively high, while the mesomorphic factor was relatively low, and the ectomorphic factor was almost the same.CONCLUSION: The somatotype of Han children in rural area in Huludao City develops better. Their heights are tall, whereas their skeletal muscles are not so muscular. Somatotype development of children is diversified with age.

AIM: To investigate the role of post-hemorrhagic shock mesenteric lymph (PHSML) in the enhancementof vascular permeability .METHODS: Eighteen Wistar rats were randomized into sham group , shock group,and shock plus mesenteric lymph drainage (shock +drainage) group.The rats in shock group and shock +drainagegroup were routinely subjected to hemorrhagic shock and hypotension [(40 ±2) mmHg] was maintained for 90 min, andthen the fluid resuscitation was performed.Mesenteric lymph was drained in the rats in shock +drainage group from resuscitationfinished to 6 h, for the observation of PHSML drainage on the vascular permeability in multiple tissues of hemorrhagicshock rats.Afterwards, human umbilical vein endothelial cells (HUVECs) were incubated with the PHSML in vitro to observethe effects of PHSML on the morphology and permeability of HUVECs .RESULTS: The degree of blue color and concentrationsof Evens blue in the lung, myocardium, kidney, liver, spleen and small intestine were significantly increased inthe shocked rats than that in sham group, while the ratios of the dry weight to the wet weight were decreased .The mesentericlymph drainage reversed these changes .Meanwhile, 4% and 10% of PHSML at 0 ~3 h and 3 ~6 h after resuscitation,and lipopolysaccharide (10 mg/L) all caused the damage of HUVECs, decreased the viability and trans-endothelial electricalresistance of HUVECs, and increased the permeability of HUVECs to fluorescein isothiocyanate -labeled albumin. CONCLUSION: PHSML is a vital factor in the enhancement of vascular permeability .

Objective To analyze the differences of size and charge heterogeneities between origi-nal humanized anti-TNF-αantibody and four similar biotherapeutic products ( SBP ) .Methods The size exclusion chromatography ( SEC-HPLC ) and weak cation exchange chromatography ( WCX-HPLC ) were used to analyze the size and charge heterogeneities , respectively.Carboxypeptidase B (CpB) treatment was employed to analyze the source of charge heterogeneity of the antibody products .Results Four SBPs showed the same pattern with the originator in SEC-HPLC, and no significant difference with the percentage of mono-mer was observed .The percentages of the aggregates of SBP-3 and SBP-4 were a little higher than those of the originator .The charge distribution of SBPs was significantly different from the originator ′s, especially in the basic region .The results from the samples treated with CpB indicated that the difference of charge distri -bution in the basic region might be caused by the C-terminal lysine variants .Conclusion Four SBPs showed similar size heterogeneity with the originator , but significant differences with charge heterogeneity were observed among them .The study suggested that more attention should be paid to the charge heterogene -ity analysis of the biosimilar products .

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.

Ginsenoside Rb3 (GRb3) is one of the main components in plasma of Panax quinquefolius Saponin of stem and leaf (PQS), which can be into human plasma. Previous studies have found PQS has estrogen-like vascular protective effects. In the present study, we investigated the estrogen-like protective effect of GRb3 on oxidative stress and dysfunction of endothelial cells induced by oxidized low-density lipoprotein. The activities of SOD, NOS and the contents of MDA in the cell lysate were examined by enzyme method or spectrophotometry. The NO and ET-1 concentrations in the cell culture supernatant were measured by ELISA method. The iNOS and eNOS mRNA expression were measured by real time RT-PCR, while the phosphorylation levels of Akt was measured by Western blotting. The results showed that GRb3 could enhance the activity of SOD, reduce the content of MDA, increase the level of NOS, NO, ET-1 and iNOS mRNA expression while decrease the eNOS mRNA expression and the phosphorylation level of Akt. These effects were blocked by estrogen receptor antagonist ICI182780. GRb3 can play a role in protecting vascular endothelial cells by estrogen receptors, the protective mechanism is similar to 17-β estrodiol.

Objective To establish a HPLC method for simultaneous determination of seven active components in Fufang Danshen tablets and Fufang Danshen dripping pills. Methods These seven compounds were analyzed simultaneously with a Zorbax C18 column by gradient elution using acetonitrile-0. 1% phosphoric acid solution as mobile phase, the flow rate was 1 mL·min-1 and the detection wavelength was set at 203, 270 and 281 nm, respectively. Results All the seven components showed good linear relation between peak area and concentration of the test, and the average recoveries were between 95. 1%-100. 4%. Tanshinone ⅡA was not detected in samples of dropping pills. Conclusion The HPLC method to determine the components including tanshinone ⅡA, salvianolic acid B, propanoid acid, protocatechuic aldehyde, notoginsenoside R1 , ginsenoside Rg1 and ginsenoside Rb1 of the two different Danshen preparations has been established, and it has the advantages of simplicity, high precision, good repeatability, and can be used for the quality control of two kinds of Fufang Danshen preparations. The content of tanshinone Ⅱ A in Fufang Danshen tablet was distinctly higher than that of dropping pills.

Objective To investigate the molecular mechanisms of effect of glycogen synthase kinase -3beta( GSK3β) on proliferation of human osteosarcoma cells .Methods Normal osteoblast hFO and osteosarcoma cell lines were examined for GSK 3βexpression and activity by Western blot and in vitro kinase assay ( NIRKA) . The effects of small molecule GSK3βinhibitors on cell proliferation and apoptosis were examined .Depletion of en-dogenous GSK3βby GSK 3βsiRNA detected the expression and phosphorylation of p 27 and its downstream cy-clinD1-CDK-Rb pathway factor by Western blot .Human osteosarcoma cell xenografts ,in athymic mice model , were treated with DMSO as control or with GSK 3βinhibitor SB-216763 or AR-A014418 by intraperitoneal in-jection,3 times a week.The tumor growth and body weight were observed in nude mice .Results Osteosarcoma cell lines showed increased GSK 3βexpression and decreased serine 9 phosphorylation compared with normal oste-oblast cells.Inhibition of GSK3βresulted in attenuated cell proliferation and increased apoptosis in most osteosar-coma cell lines in vitro and in vivo in MG 63 xenografts in rodents but not in hFOB cells .We decreased endoge-nous GSK3b activity,tumor growth was inhibited in SB216763,AR -A014418 group compared with control group.There was statistical significance(P<0.05).GSK3βinhibition in osteosarcoma cells was associated with decreased p27 expression, Rb expression and phosphorylation level of decline , CDK2, 4, 6 protein level de-creased,the upregulation of cyclin D1 expression but the phosphorylation level of no effect .Conclusion In this research,we demonstrate that deregulated GSK 3βsustains osteosarcoma cells survival through modulation of p27and cyclinD1-CDK-Rb pathway.The result will open up a potential target for clinical treatment of osteosar -coma.