BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.

Objective To evaluate the safety and effectiveness of the treatment by selective β 1 receptor blockers on patients with chronic obstructive pulmonary disease (COPD).Methods Eighty cases of COPD Ⅲ (stable period) inpatient with or without coronary heart disease were collected in The Second People's Hospital of Shanghai from September 2012 to November 2013.The patients were randomly divided into testing group (Metoprolol treatment group) and control group (regular treatment group) with 40 cases for each group.Metoprolol group therapy based on the use of conventional metoprolol tablets,an initial dose of metoprolol 12.5 mg/d,titrated to the appropriate dose based on heart rate and tolerance of the morning resting heart rate of 55 to 60 times/min that reached the target dose of metoprolol continuous medication for 12 months.Blood gas analysis were recorded before and after treatment,pulmonary function,and 6 min walk test (6MWT) and were chronic lung disease Assessment Test (CAT) Rating.The control group was administrated regular treatment while the testing group added small dose of Metoprolol with titration to an appropriate dose on this basis.12 months in a row,and assessed the end stage.Results (1) After the application of selective β receptor blockers on testing group,no statistically significant difference (P＞0.05) in the values of FEV1 in anticipation value％ (testing group:(45.45 ± 4.68) ％ vs.(43.32 ± 4.84) ％;control group:(44.23 ± 4.68) ％ vs.(42.58 ±4.24)％),PaO2(testing group:(75.92± 10.78) mmHg vs.(74.86± 11.21) mmHg;control group:(70.23 ±6.45) mmHg vs.(72.36±7.28) mmHg) and PaCO2(testing group:(46.28±8.28) mmHg vs.(47.46±10.22) mmHg);control group:(44.54 ± 8.89) mmHg vs.(42.36 ± 7.45) mmHg) before and after treatment.But the 6MWD (testing group:(287 ± 23) m vs.(384± 34) m;control group:(284 ± 25) m vs.(295 ±21) m) and COPD appraisal test(CAT) (testing group:(21±7) score vs.(17±6) score);control group:(22 ±5) score vs.(20± 6) score) had improved significantly compared with that before treatment,with significant difference(t=4.903,4.784;P＜0.05).Conclusion Selective β receptor blockers have no effect on the airway resistance of COPD patients and reduction on pulmonary function.It can also increase the exercise tolerance and enhance the living quality for improving clinical prognosis.

Objective To evaluate the efficiency of second trimester screenings for Down syndrome using alpha-fetoprotein and β-human chorionic gonadotropin duplex.MethodsPregnant women of south Zhejiang were screened for Down syndrome fetuses by maternal alpha-fetoprotein and β-human chorionic gonadotropin duplex during second trimester.The high-risk women underwent prenatal diagnosis by amniocentesis,cell culture and chromosome analysis.The newborns followed up by the maternal and child tertiary health care network and suspected to have Down syndrome were diagnosed by peripheral blood chromosome analysis.Statistical analysis was performed using two-sample t test and x2 test.Risk probability of Down Syndrome was calculated by random screening software. Results From Oct.2007 to May 2009,1130 of 32 188 singleton pregnant women in second trimester received prenatal screening were discovered with high risk(≥1 ∶ 270).Prenatal diagnosis was performed in 90.79％ cases (1026/1130) of high risk women and seven fetuses were diagnosed as Down syndrome by amniotic fluid chromosome analysis,and the pregnancies were terminated.Among the other 104 cases without prenatal diagnosis one Down syndrome baby was delivered.Six of 31 058 pregnancy women with low risk delivered Down syndrome babies with the incidence of Down syndrome of 0.19‰ (6/31 058).Detection rate of second trimester screenings for Down syndrome using alpha-fetoprotein and β-human chorionic gonadotropin duplex was 57.14％(8/14).False positive rate was 3.48％ (1122/32 188).Positive predictive value was 7.08‰(8/1130).During the same period,there were 23 813 pregnant women who didn't receive screening and 15 fetuses with Down syndrome were diagnosed after birth.There was no statistical difference in the prevalence rate of Down syndrome between those pregnant women who received prenatal screening or not [0.43‰ (14/32 188) vs 0.63‰ (15/23 813),x2 =1.004,P＞0.05].The prevalence of Down syndrome was 0.52‰ (29/56 001) in this area. ConclusionsThe prenatal screening and diagnosis could reduce the birth rate of Down syndrome patients.However,detection rate,false positive rate and positive predictive value of which were lower than reports in other studies.It's possible that the reference data might be not suitable for Chinese.

BACKGROUNDCathepsin S and its endogenous inhibitor cystatin C are implicated in the pathogenesis of atherosclerosis, especially in the plaque destabilization and rupture leading to acute coronary syndrome. However, whether circulating cathepsin S and cystatin C also change in association with coronary plaque morphology is unknown yet.

METHODSWe recruited 98 patients with unstable angina (UA, n = 6) or stable angina (SA, n = 2) who had a segmental stenosis resulting in > 20% and < 70% diameter reduction in one major coronary artery on coronary angiography. Thirty-one healthy subjects served as controls. Intravascular ultrasound (IVUS) was used to evaluate plaque morphology. Plasma cathepsin S and cystatin C were measured as well.

RESULTSAt the culprit lesion site, plaque area ((7.85 +/- 2.83) mm(2) vs (6.53 +/- 2.92) mm(2), P = 0.027), plaque burden ((60.92 +/- 11.04)% vs (53.87 +/- 17.52)%, P = 0.025), remodeling index (0.93 +/- 0.16 vs 0.86 +/- 0.10, P = 0.004) and eccentricity index (0.74 +/- 0.17 vs 0.66 +/- 0.21, P = 0.038) were bigger in UA group than in SA group. Plasma cathepsin S and cystatin C were significantly higher in patients than in controls (P < 0.01). Plasma cathepsin S was higher in UA group ((0.411 +/- 0.121) nmol/L) than in SA group ((0.355 +/- 0.099) nmol/L, P = 0.007), so did the plasma cystatin C ((0.95 +/- 0.23) mg/L in UA group, (0.84 +/- 0.22) mg/L in SA group; P = 0.009). Plasma cathepsin S positively correlated with remodeling index (r = 0.402, P = 0.002) and eccentricity index (r = 0.441, P = 0.001), and plasma cystatin C positively correlated with plaque area (r = 0.467, P < 0.001) and plaque burden (r = 0.395, P = 0.003) in UA group but not in SA group.

CONCLUSIONSPlasma cathepsin S and cystatin C increased significantly in UA patients. In angina patients, higher plasma cathepsin S may suggest the presence of vulnerable plaque, and higher plasma cystatin C may be a clue for larger atherosclerotic coronary plaque.

Adult ; Aged ; Aged, 80 and over ; Cathepsins ; blood ; Coronary Artery Disease ; blood ; diagnostic imaging ; pathology ; Cystatin C ; blood ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Ultrasonography, Interventional ; methods

BACKGROUNDThere is no consensus regarding the performance for endorectal ultrasonography (ERUS) at every stage of rectal cancer. Thus, the purpose of our study was to further assess the value of ERUS in the preoperative staging of rectal cancer.

METHODSA retrospective study was performed with 44 consecutive patients (mean age: (63.3 ± 10.2) years) who underwent surgical treatment for endorectal carcinoma and were preoperatively evaluated using Biplane ERUS between September 2008 and December 2010. We compared the ERUS staging with the pathological findings based on surgical specimens.

RESULTSERUS staging agreed with the histologic staging in 39 of the 44 (88.6%) patients: the agreement on the depth of transmural invasion was good (κ = 0.73; 95%CI: 0.60 - 0.86, P = 0.000). The detection sensitivities of rectal cancer with ERUS were as follows: T1 85.7%, T2 87.5%, T3 88.9%, and T4 100.0% with specificity values of T1 97.3%, T2 92.9%, T3 96.2%, and T4 97.6%. ERUS correctly staged patients with T1 95.5%, T2 90.9%, T3 70.5%, and T4 97.7%. The positive predictive value of ERUS was lowest for T4 (75%), but highest for T3 (94.1%) followed by T2 (87.5%) and T1 (85.7%); the negative predictive values of ERUS from high to low were ordered as T4 (100%), T1 (97.3%), T2 (92.9%), and T3 (92.6%). The percentage of total over-staged cases was 4.5% and the under-staged cases was 6.8%. The extent of perirectal lymph node metastases was determined with a sensitivity of 68.4% (13/19), specificity of 80.0% (20/25), and diagnostic accuracy of 75.0% (33/44).

CONCLUSIONBiplane ERUS has a high diagnostic accuracy for tumoral invasion of the rectal wall at every T stage, but relatively low diagnostic accuracy for lymph node metastases.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Rectal Neoplasms ; diagnostic imaging ; pathology ; Retrospective Studies ; Ultrasonography

OBJECTIVE To observe whether human CD4 + T cells could be activated by immuno-globulin D (IgD) via IgD receptor(IgDR)-Lck. METHODS Human CD4+ T cells were purified from peripheral blood mononuclear cells (PBMCs) with microbeads. The viability of T cells were detected by CCK-8. The binding affinity and expression of IgDR on T cells were detected by flow cytometry. The protein expression of IgDR, Lck and P-Lck were analyzed by western blot. RESULTS IgD could concentration-dependent bind to IgDR on CD4+ T cells. The expression of IgDR was increased in response to treatment with IgD in a time- dependent and concentration- dependent manner. Stimulating by IgD resulted in enhanced phosphorylation of Lck compared with that in the medium control sample. The expression of Lck was not changed. As inhibitor of PTK, Herbimycin A or A770041, which combined with IgD could significantly inhibit phosphorylation of Lck(Tyr394). The proliferation promoting effect of IgD was blocked by Herbimycin A or A770041. IgD could stimulate CD4+ T cell activation and proliferation through upreg?ulating activating tyrosine residue of Lck (Tyr394) phosphorylation. CONCLUSION These results demon?strate that IgD exaggerates CD4+T cell activities, which may be through promoting Lck phosphorylation.

OBJECTIVETo determine the expression level changes of survivin, a inhibitor of apoptosis protein, followed by activation of insulin receptors in human hepatocellular carcinoma HepG2 cell line, and to investigate the signalling pathway involved in the regulation.

METHODSHuman hepatocellular carcinoma HepG2 cells were treated with insulin alone or pre-treated with LY294002, a specific inhibitor of PI3K signalling pathway, to determine whether blocking PI3K signaling can attenuate the up-regulation of survivin expression. Real time RT-PCR and Western blot analysis were used to measure survivin mRNA and protein changes before and after treatment, respectively.

RESULTSWithout serum supplement, HepG2 cells expressed a small amount of survivin. Insulin induced survivin expression in a dose- and time-dependent fashion. Survivin expression was blocked if cells were pre-treated with LY294002 prior to insulin stimulation.

CONCLUSIONInsulin induces survivin expression via PI3K signalling pathway, suggesting that to interfere the key gene in this signalling pathway may block survivin expression, therefore, promoting apoptosis in hepatocellular carcinoma cells.

Apoptosis ; Chromones ; pharmacology ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; Insulin ; administration & dosage ; pharmacology ; Microtubule-Associated Proteins ; genetics ; metabolism ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; RNA, Messenger ; metabolism ; Signal Transduction ; Up-Regulation

To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.
Animals ; Brain Ischemia ; drug therapy ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

Adult ; Antigens, CD34 ; Antiviral Agents ; therapeutic use ; Bone Marrow Cells ; virology ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; genetics ; physiology ; Humans ; Immunoglobulins ; therapeutic use ; Lamivudine ; therapeutic use ; Liver Transplantation ; Male ; Middle Aged ; Monocytes ; Postoperative Period ; Virus Replication

OBJECTIVETo investigate the inhibitory effect of lycium pigment on lipopolysaccharide (LPS)-induced uveitis in rats and its mechanism.

METHODThe rat uveitis model was established by 30-day oral administration of lycium pigment (50, 100, 200 mg x kg(-1)) and footpad injection of LPS. Ocular tissues were collected for a histopathological inspection. The protein, nitric oxide and ADMA in aqueous humor, level of inducible nitric oxide synthase (iNOS) in retina, activities of serum total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), and content of malondialdehyde (MDA) were determined by using Western blot, ELISA and biochemical methods.

RESULTAccording to the pathological study, lycium pigment (50, 100, 200 mg x kg(-1)) could notably reduce the inflammatory cell infiltration around corpus ciliare matrix of uveitis rats, and the concentration of protein and nitric oxide, and increased ADMA in aqueous humor. Lycium pigment (100, 200 mg x kg(-1)) could significantly inhibit the expression of iNOS in ocular tissues. In addition, lycium pigment (100, 200 mg x kg(-1)) also decrease the activities of serum T-AOC, SOD, GSH-PX, and the content of lipid peroxide MDA.

CONCLUSIONLycium pigment has the inhibitory effect on LPS-induced uveitis in rats. Its mechanism is related to the regulation of nitric oxide/ADMA pathway and the improvement of oxidation resistance.

Animals ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Lycium ; chemistry ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Pigments, Biological ; administration & dosage ; Plant Extracts ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Uveitis ; chemically induced ; genetics ; metabolism ; prevention & control