Objective To establish antibody titre quantitation method by ELISwithoustandard substance .MethodThe tesgroupof non-specifi,negative control ,specifiand total antibodiewere sefodetecting serantibodieby ELIS.Aftelineafitting of the time-absorbance datof early developing ,the fitted slopewere used athe velocity of absorbance changing , which were denoted by ν0 ,νC,νS,νT.Based on thathe ν-value and the concentration ? of determinand were linearelationship while the substrate waexcessive ,the function with parameteof ν-valueforeflecting the multiple proportionof antibodiecon-centrationbetween specifiand negative control groupcould be deduced .The assessmenof specifiIgG antibodiein serof KM mice immunized with fish collagen waused aan instance .ResultThe function C/C= (ν-ν0 )/(νC-ν0 ) could calculate the mul-tiple proportion(titre) of antibodieconcentrationbetween specifiand negative control group.Conclusion The above method of antibody titre quantitation isuitable fosemi-quantitative analysiwithoustandard substance .

ObjectiveTo find out if the immune system derived thyroid stimulating hormone(TSH) β splice variant(TSHβ-Ⅴ) would be regulated by circulating thyroid hormone levels to get a further understanding of the function and mechanism of this TSHβ-Ⅴ in thyroid homeostasis.MethodsA total of 20 weaning Balb/c mice (half male and half female) were selected and randomly divided into two groups according to their body mass and gender(n =10).Mice of control group were fed with common diet and deionized water.Mice of the low-iodine(LI) group were fed with low-iodine diet(containing iodine 20 - 40 μg/kg,iodine-intake about 0.25 μg/d) and deionized water.The experimental period was 3 months.At the end of the experiment,mice were executed and the blood was collected to observe the levels of TSH and thyroid hormone by chemiluminescence immunoassay (CIA) ; bone marrow (BM),peripheral blood(PBL),thyroid gland and pituitary were collected to assay the TSHβ-Ⅴ mRNA expression by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR).ResultsThe serum free thyroxine(FT4) and total thyroxine(TT4) levels in LI group of mice[(0.47 ± 0.70)nmol/L,(2.41 ± 0.28)pmol/L] were significantly lower than that of the control group of mice [(55.2 ± 3.68) nmol/L, (32.72 ± 1.02) pmol/L,t =43.81,86.04 、all P ＜ 0.01 ] and the serum total triiodothyronine(TT3) and free triiodothyronine(FT3) reduction in LI group of mice[ (0.76 ± 0.08)nmol/L,(4.01 ± 0.40)pmol/L] were significantly lower than that of the control group of mice [ (1.10 ± 0.06)nmol/L,(5.40 ± 0.38)pmol/L,t =9.81,7.5 1,P ＜ 0.01 ].Iodine insufficiency strongly elevated the serum TSH in LI group of mice[ (35.67 ± 17.39)mU/L] than that in control group of mice[ (0.24 ± 0.10)mU/L,t =- 6.11,P ＜ 0.01 ].The mRNA levels of TSH β-Ⅴ in BM (9.62 ± 0.60) and in PBL( 9.25 ± 0.83 ) of LI group of mice were lower than those in control group of mice (7.69 ± 0.36,7.11 ± 0.41,t =6.77,5.64,P ＜ 0.01),while the mRNA level of TSH β-Ⅴ in pituitary of LI group of mice (1.99 ± 0.61) was increased compared with that in control group of mice (5.75 ± 0.98,t =- 8.02,P＜ 0.01).Compared with control group of mice(9.12 ± 0.62),the level of thyroid TSH β-Ⅴ mRNA in LI group of mice (9.32 ± 0.91 ) was not significantly changed (t =0.45,P ＞ 0.05).There was no detectable native TSHβ in BM,PBL and thyroid.The mRNA level of native TSHβ in pituitary in LI group of mice( - 7.17 ± 1.78) was dramatically elevated compared to that in control group of mice( - 1.43 ± 0.51,t =- 7.60,P ＜ 0.01 ).ConclusionsThe mRNA levels of TSHβ-Ⅴ are suppressed in BM and PBL in low iodinediet induced hypothyroidism mice,which suggest that immune system derived TSHβ-Ⅴ may be more important thannative TSHβ in immune-thyroid regulation.

Objective To revise the Symptom Distress Scale for postoperative patients with pituitary tumor and to test its reliability and validity. Methods On the base of previous qualitative interview and literature review, Delphi consultation was performed to identify items of the Symptom Distress Scale for postoperative patients with pituitary tumor. By convenience sampling method, totally 191 patients from four first-class ternary hospitals in Jiangsu province were investigated effectively by this scale. Results A scale of 4 factors and 16 items was identified by expert interviews, item analysis, exploratory factor analysis and the four factors could explain 69.812%of the variance. The Cronbachαcoefficient of the scale was 0.920, the content validity index was 0.915, and the interrater reliability was 0.860. Conclusions Symptom Distress Scale for postoperative patients with pituitary tumor has good reliability and validity to assess the symptom distress of pituitary tumor patients after operation.

OBJECTIVE To investigate the clinical significance of human papillomavirus in paraffin-embedded cervical cancer and precancerous lesion tissue by gene clip technology.METHODS 153 Patients with paraffin-embedded examples.DNA was extracted from paraffin-embedded tissues and amplified by polymerase chain reaction(PCR).RESULTS The positive rate of high-risk HPV of inflammation was 8.33%,CINⅠ45.83%,CINⅡ/CINⅢ 87.50% and invasive cancer 92.21%.The HPV infection rate of squamous cell carcinoma was 94.12%.The HPV infection rate of adenocarcinoma was 88.46%.Among all the patients with cervical cancer and CIN,the infection rate of HPV16,the most genotype,was 88.98%.The infection rate of HPV18,the second most subtype,was 33.06%.In addition,the minority were infected HPV52、33、59、68.Among 48 cases of cervical squamous cell carcinoma,the infection rate of HPV16,HPV18 was 93.73% and 27.08% respectively.Among 23 cases of cervical adenocarcinoma,the infection rate of HPV16,HPV18 was 82.61% and 52.17% respectively.On the other hand,all the patients with cervicitis were HPV single infection.The HPV multiple infection rate of CINⅠ,CINⅡ/CINⅢ,cervical cancer was 20.00%,28.57%,36.62% respectively.CONCLUSIONS Gene chip technology can detect multiple HPV genotypes in paraffin-embedded tissues with high sensitivity and specificity,which is useful in the pathogenesis and prevention of cervical cancer.

OBJECTIVETo compare the therapeutic effects of acupuncture and Motilium on diabetic gastroparesis (DGP).

METHODSSeventy-two cases of DGP were randomly divided into a treatment group and a control group. The treatment group were treated with needling method for harmonizing spleen-stomach as main, with Quchi (LI 11), Hegu (LI 4), Zhongwan (CV 12), Zusanli (ST 36), Fenglong (ST 40), Yinlingquan (SP 9), Sanyinjiao (SP 6), Xuehai (SP 10), Diji (SP 8), and others selected, twice each day, 10 days constituting one course, with an in terval of 2 days; the control group were treated with oral administration of Motilium 10 mg, 3 times each day, 30 min before meals.

RESULTSThe total effective rate of 91.7% in the treatment group was better than 77.8% in the control group (P < 0.05).

CONCLUSIONThe therapeutic effect of the needling method for harmonizing spleen-stomach on diabetic gastroparesis is better than that of Motilium.

Acupuncture Therapy ; methods ; Adult ; Diabetes Complications ; therapy ; Female ; Gastroparesis ; therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology

OBJECTIVETo investigate the changes of NF-kappa B binding activity, the expression of PPARr and their correlation in the liver of rats with fatty liver disease (FLD) induced by different pathogenic factors and to investigate the molecular mechanism of the inflammation in FLD.

METHODS40 Wistar rats were randomly divided into 4 groups of ten each: normal group, alcohol group, fat-rich diet group, alcohol adding fat-rich diet group. The rats were sacrificed at the end of the 16th week from the starting day of the experiment. Serum and liver specimens were collected. Histological specimens were stained with HE, SudanIV, and Masson and then studied microscopically. The ultrastructural changes were also checked under an electron microscope. NF-kappa B binding activity and the expression of PPARr mRNA were determined by electrophoretic mobility shift assay (EMSA) and RT-PCR respectively. The correlations between NF-kappa B binding activity and the expression of PPARr and the biochemical indexes were analyzed.

RESULTSSteatosis, inflammation, necrosis and fibrosis were present in livers of the rats of all the experimental groups, and were most severe in the alcohol adding fat-rich diet group. NF-kappa B binding activity was markedly increased in the livers of the alcohol group (142+/-16.32) and of the alcohol adding fat-rich diet group (238+/-19.14) in comparison to the livers of the normal (73+/-9.24, F = 6.36, 17.93) and those of the fat-rich diet group (84+/-10.38, F = 5.96, 16.20). Binding activity was higher in the alcohol adding fat-rich diet group than that in the simple alcohol group, but there was no difference between those of the fat-rich diet and normal groups. The level of PPARr mRNA was lower in the livers of the alcohol, fat-rich diet, alcohol adding fat-rich diet groups (0.2530+/-0.069, 0.3647+/-0.082, 0.1226+/-0.054) than that of the controls (0.8097+/-0.094) (F = 15.43, 7.24, 21.45). NF-kappa B binding activity was correlated positively with the level of serum TNF alpha (r = 0.527, 0.639) and the content of MDA in the liver homogenates (r = 0.723, 0.537), but negatively with the expression of PPARr in the livers of the alcohol and the alcohol adding fat-rich diet groups (r = -0.568, -0.891).

CONCLUSIONThe enhanced nuclear factors NF-kappa B binding activity and decreased expression of PPARr play a pivotal role in the inflammatory response of FLD induced by alcohol and fat-rich diet. It may provide a new idea for treating FLD effectively.

Animals ; Fatty Liver ; metabolism ; Liver ; metabolism ; Male ; NF-kappa B ; metabolism ; PPAR gamma ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

OBJECTIVETo investigate the distribution of ER isoforms in endometriosis and eutopic endometrium.

METHODSTissue samples of patients with ovarian endometriosis, treated in People's Liberation Army General Hospital from January 2004 to December 2006, were retrieved. A total of 60 cases of ovarian endometriotic cysts with their corresponding eutopic endometrium (30 cases of proliferation phase and 30 of secretary phase eutopic endometrium) and 30 cases of normal endometrium (15 proliferative and 15 secretary phase endometrial samples respectively) were included. Expressions of ERalpha and ERbeta were analyzed using immunohistochemistry and the expression ratio was statistically analyzed by using SPSS 12.0 software.

RESULTSExpressions of both ERalpha and ERbeta in epithelial cells were positively correlated with that of the stromal cells. The expression of ERalpha in eutopic endometrium (73.3% in epithelium and 76.7% in stroma) was significantly higher than that in ovarian endometriotic cysts (43.3% in epithelium and 46.7% in stroma), or normal control (56.7% in epithelium and 50.0% in stroma, respectively, each P < 0.05. However, the expression of ERbeta (90.0% in epithelium and 76.7% in stroma) was higher in ovarian endometriotic cysts than that in the eutopic endometrium (68.0% in epithelium and 63.3% in stroma respectively, P < 0.05), and ERbeta expression in eutopic endometrium was higher than that in the normal control endometrium (36.7% in epithelium and 26.7% in stroma, respectively, P < 0.05). The expressions of both ERalpha and ERbeta changed periodically in eutopic and normal endometrium, whereas ERalpha and ERbeta level were less variable in the ectopic endometrium. The expression of ERbeta was statistically higher than that of ERalpha (P < 0.05) in ectopic endometrium, whereas no significant difference was seen between the two isoforms in the eutopic or normal endometrium.

CONCLUSIONSBoth ERalpha and ERbeta have higher expression levels in eutopic endometrium of patients with ovarian endometriotic cysts. ERbeta is predominantly expressed in endometriotic cysts, where the expression of ERalpha is limited. The different distribution of ERalpha and ERbeta may play an important role in the development of ovarian endometriosis.

Adult ; Choristoma ; pathology ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Epithelium ; Estrogen Receptor alpha ; analysis ; Estrogen Receptor beta ; analysis ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Protein Isoforms ; analysis ; Receptors, Estrogen ; analysis ; Stromal Cells ; metabolism

This study was aimed to investigate the effects of glycyrrhetinic acid (GA) on proliferation, apoptosis and survivin mRNA expression in human myeloma cell line U266 in vitro. Cell proliferation was assayed by MTT method. Both cell apoptosis and cell distribution in cell cycle were analyzed by using flow cytometry. Scanning electron microscopy was used to observe the morphological changes in U266 cells induced by GA. Expression of survivin mRNA was detected by quantitative real-time reverse transcription-polymerase chain reaction. The results showed that GA inhibited proliferation of U266 cells in a time- and dose-dependent manners in vitro. GA presented apoptosis induction potency to U266 cells, obvious changes in morphology of U266 cells was observed under scanning electron microscope. The cells were arrested at the G(0)/G(1) phase, showing the accumulation in G(0)/G(1) phase, reduction of cells in G(2)/M phase and S phase. GA could down-regulate the expression of survivin gene in U266 cells in a dose-dependent manner. It is concluded that GA can inhibit proliferation of U266 cells in a time- and dose-dependent manners and induce apoptosis of this cell line in vitro through arresting G(0)/G(1) phase and down-regulating expression of survivin.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Glycyrrhetinic Acid ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology

OBJECTIVETo study the role of serum 25-hydroxyvitamin D in the early diagnosis of vitamin D deficiency rickets.

METHODSConcentrations of serum 25(OH)D, calcium, phosphorus and alkaline phosphatase were measured in normal control (n=73), suspected rickets (n=45) and confirmed rickets groups (n=65). Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of serum 25(OH)D for rickets.

RESULTSSerum 25(OH)D levels in the suspected and confirmed rickets groups were 83±30 and 72±31 nmol/L respectively, which was lower than in the normal control group (112±37 nmol/L) (P<0.01). There was no significant difference between the suspected and confirmed rickets groups (P>0.05). Vitamin D deficiency rates in the suspected and confirmed rickets groups were higher than in the control group (P<0.01). The ROC curve area of serum 25(OH)D for the diagnosis of rickets was 0.760 (95%CI 0.692-0.820, P<0.01), and the optimal operating point was 90.70 nmol/L (sensitivity 68.49%, specificity 72.73%). There was no significant difference in levels of calcium, phosphorus and alkaline phosphatase between the three groups (P>0.05).

CONCLUSIONSSerum 25(OH)D levels in infants with suspected and confirmed rickets are significantly reduced and this may reflect vitamin D deficiency . Therefore, it may be useful to check serum 25(OH)D levels in screening for rickets.

Female ; Humans ; Infant ; Male ; ROC Curve ; Rickets ; blood ; diagnosis ; Vitamin D ; analogs & derivatives ; blood