factors for quality of life in patients over 12 months after diagnosis of non-small cell lung cancer / 上海交通大学学报（医学版）

Objective • To examine quality of life and its influencing factors in patients over 12 months after diagnosis of non-small cell lung cancer (NSCLC). Methods • A total of 111 patients included in the study completed the self-reported measures. Quality of life was measured using Functional Assessment of Cancer Therapy-Lung. Other measures included socio-demographic characteristics, disease- and treatment-related information, and emotional feelings. Results • The factors influencing the quality of life of NSCLC patients were investigated from two approaches: treatment and subjective feeling. Univariate Logistic regression analysis found the risk factors for low quality of life included sense of low self-esteem and loneliness, bone metastasis, pathological type, chemotherapy cycle, age, cohabitant type and financial burden. Conclusion • The factors associated with quality of life in lung cancer patients are diverse and extensive, and it warrants attention on psychological support and symptom management for these patients.

Establishment and characterization of A549 tumor monoclonal cell line with UCHL1 gene deletion / 生理学报

The purpose of this study was to investigate the effects of ubiquitin C-terminal hydrolase-L1 (UCHL1) on non-small cell lung cancer cell line A549. UCHL1 gene knockout A549 cell line was constructed by CRISPR-CAS9 gene editing technique. The mRNA and protein levels of UCHL1 were examined by RT-PCR and Western blot, respectively. Cell proliferation and cycles were analyzed by CCK-8 method and flow cytometry, respectively. The sensitivity of A549 cells to cisplatin was detected by CCK-8 method. Migration ability of A549 cells was detected by scratch assay and Transwell test, and p-Erk expression level was assessed by Western blot. The results showed that UCHL1 gene knockout A549 cells were successfully constructed by CRISPR-CAS9 gene editing technique. After UCHL1 gene knockout, there was no significant change in cell proliferation and cell cycle ratios in A549 cells. UCHL1 gene knockout A549 cells exhibited decreased sensitivity to cisplatin and migration activity, as well as increased p-Erk expression level. These results suggest that the loss of UCHL1 gene function may reduce the sensitivity and migration ability of A549 cells, and this effect may be related to the activation of Erk1/2 signaling pathway.

Impact of multifactor intensive intervention on self management, risk factor control and outcome of post percutaneous transluminal coronary intervention patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To test the efficacy of multifactor intensive intervention for post percutaneous transluminal coronary intervention (post-PCI) outpatients on self management, risk factor control and outcome.</p><p><b>METHODS</b>A total of 263 patients with coronary heart disease (CAD) discharged from our cardiac center were randomized into usual care (4 CAD lectures focusing on the 2(nd) CAD prevention and patients-oriented outpatient visit) and intensive intervention (4 CAD lectures focusing on the 2(nd) CAD prevention, CAD outpatient visit twice a month, monthly telephone instructions on risk factor control and optimal medication). Patients were followed for 12 months and 250 patients completed follow-up.</p><p><b>RESULTS</b>There were more patients achieved a LDL-C level of less than 2.6 mmol/L in intensive intervention group than in usual care group (71.2% vs. 48.3%, P < 0.01). The percentages of patients taking dietary control (55.3% vs. 26.2%, P < 0.01) and physical exercises (64.4% vs. 39.0%, P < 0.01), receiving beta-adrenergic receptor blocker (75.0% vs. 50.8%, P < 0.01) and statins (72.0% vs. 54.2%, P < 0.01) were significantly higher while cardiovascular event rate (5.9% vs. 0%, P = 0.005)was significantly lower in intensive intervention group than in usual care group.</p><p><b>CONCLUSION</b>Multifactor intensive intervention is helpful on improving the second prevention for post-PCI coronary heart disease patients.</p>

Nongenomic action and mechanism of 17β-estradiol in cytosolic calcium concentration in delayed implantation mouse endometrial stromal cells / 生理学报

To investigate the existence of nongenomic action of 17β-estradiol (E(2)) in the delayed implantation mouse endometrial stromal cells, the changes in intracellular calcium concentration ([Ca(2+)](i)) and the upstream of calcium signal in vitro were detected. The experiment was composed of two parts. Firstly, the change in [Ca(2+)](i) in endometrial stromal cells induced by E(2) under different conditions was detected. The mice were divided into 6 groups as follows: 0.1% dimethylsulfoxide (DMSO) control group, 1×10(-8) mol/L bovine serum albumin (BSA) control group, 1×10(-8) mol/L E(2) group, 1×10(-8) mol/L E(2) conjugated with BSA (E(2)-BSA) group, 1×10(-8) mol/L E(2) + calcium-free medium group, 1×10(-8) mol/L E(2) + 5 mg/mL tamoxifen group, with 4 mice in each group. The endometrial tissue was obtained from delayed implantation mice at pregnant day 7, and digested by incubation of tissue minces in Hankos balanced salts (HBSS, pH 7.2), which contained glucose (1 g/L), and collagenase I (0.125%), for 1 h at 37 degrees C. The stromal cells were preloaded with 2.5 mmol/L Fluo-3/AM, a fluorescent probe of calcium, for 30 min. A confocal laser scanning microscope, which fixed the wave length of excitation and emission at 488 nm and 526 nm, respectively, was used to detect the change in [Ca(2+)](i). Secondly, the mechanism of E(2) effects in endometrial stromal cells was investigated. Immunofluorescent method was used to detect the change in phosphorylation of phospholipase C (PLC) before and after the stromal cells were treated with E(2) for 5 min, 15 min, and 30 min. Seven delayed implantation mice were used. The results were as follows. [Ca(2+)](i) increased immediately and reached the maximum at 15 min after the stromal cells were treated with 1×10(-8) mol/L E(2) and returned to the normal level at 30 min. In E(2)-BSA group and E(2) + calcium-free medium group the same results were obtained as that in E(2) group, but there was no increase of [Ca(2+)](i) in DMSO and BSA groups. Tamoxifen, a traditional antagonist of estrogen receptor, did not inhibit the increase in [Ca(2+)](i) induced by E(2). Immunofluorescent results showed that the change in phosphorylated-PLC level had the same trend as [Ca(2+)](i) after the cells were treated with E(2). Compared with that in the control group, the immunofluorescent intensity increased at the beginning and achieved the maximum at 15 min (P<0.001), then declined to the normal level at 30 min. These results suggest that the rapid response of [Ca(2+)](i) induced by E(2) in the endometrial stromal cells in delayed implantation mice is possibly carried out through a nongenomic pathway, and the transmembrane signal transduction is related to the phosphorylation of PLC in this process.

The progesterone-induced expression of cyclin G1 and its effect on the proliferation of mouse uterine epithelial cells / 生理学报

The aim of the present study is to investigate the effect of progesterone-induced expression of cyclin G1 on the proliferation of endometrial epithelial cells. To obtain mouse endometrial epithelial cells, the uteri were isolated from ovariectomized mice which were injected subcutaneously with 100 ng estradiol per day for two days. Then the uteri were digested by dispase and pancreatin respectively. Endometrial epithelial cells were cultured in DMEM/F12 containing 6% fetal bovine serum, and divided into four groups when they grew to confluence. Each of the groups was treated as follows: Group E was treated with 0.01 micromol/L estradiol only, group P was treated with 1 micromol/L progesterone, group EP was treated with both 0.01 micromol/L estradiol and 1 micromol/L progesterone, and group C was treated with 0.01% DMSO for control. Immunocytochemistry was used to examine the expression of cyclin G1 protein. MTT assay was used to evaluate metabolic activity of cells. Flow cytometry was used to check the number of cells distributing in each phase of the cell cycle. The result of immunocytochemistry showed that there was no expression of cyclin G1 protein in group C and group E, while cyclin G1 was obviously expressed in group P and group EP and localized in nucleus. In the MTT assay, compared with group C, the viability of group E significantly increased, while that of both group P and group EP decreased significantly. The results of flow cytometry were in accordance with those of MTT, which showed that compared with group C, group E had a higher proportion of cells in S phase, while group P, as well as group EP had a lower proportion of cells in S phase but a higher proportion in G1 phase and G2/M phase. These results indicate that progesterone could induce cyclin G1 expression in the primary culture of mouse endometrial epithelial cells, meanwhile inhibit the proliferation of cells and block the cell cycle progression. Thus, progesterone-induced expression of cyclin G1 is probably a negative factor in regulating cell cycle, which is involved in the inhibitory effect of progesterone on the proliferation of endometrial epithelial cells.

Immunogenicity of recombinant human zona pellucida-3 peptides expressed in E. coli and efficacy of their antisera to inhibit in vitro human sperm-egg binding / 生理学报

The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides.

Integrins mediate the increase of concentration of intracellular free calcium in mouse eggs / 生理学报

For studying the effect of integrin on the [Ca(2+)](i) of mouse eggs and its transmembrane signaling mechanism, zona-free mouse eggs were loaded with calcium probe Fluo-3/AM and the intensity of fluorescence of the eggs treated with different factors was measured through laser confocal microscopy. The results showed that the [Ca(2+)](i) of zona-free mouse eggs was increased when the eggs were treated with RGD peptide, fibronectin (Fn) and anti-mouse integrin subunit alpha(6) and beta(1) monoclonal antibodies, respectively. The [Ca(2+)](i) of the mouse eggs was also increased when the eggs were placed in calcium-free medium and treated with RGD peptide or Fn. The changes in the mouse egg [Ca(2+)](i) caused by RGD and Fn were similar to those caused by sperm. However, the concentration of Ca(2+) of the zona-free mouse eggs pretreated with tyrosine kinase inhibitor was not increased when the eggs were treated in the same way, and, neither was the intracellular calcium increased in those eggs pretreated with PKC inhibitor when the eggs were treated with RGD peptide. It is therefore suggested that the occupancy of integrins on the membrane of mouse eggs by their ligands mediates the release of Ca(2+) and then the increase in the [Ca(2+)](i) of eggs, which is one of the early events of egg activation. The tyrosine kinase signaling pathway and PKC are involved in this process as well.

Expression and purification of three fusion proteins containing a single B-cell epitope (beta5, beta9 or beta8) of human chorionic gonadotropin beta subunit / 生物工程学报

The possibility of using a subunit or fragment of human chorionic gonadotropin (hCG) as an immunogen for birth control has been actively explored for many years. This protein homone is produced by the fertilized egg and is required for implantation of the blastocyst into the maternal uterus and the maitenance of pregnancy. In previous studies, several bio-synthesized hCG chimeric peptides (CP) that contain three linear B-cell epitopes (beta5, beta9 and beta8) of beta-hCG subunit together with various foreign 'promiscuous' T-cell epitopes were constructed and expressed as potential new hCG vaccine immunogens. In order to detect antibodies to each of the individual B-cell epitopes present in the animal antiserum raised against the hCG CPs, we decided to construct three recombinant proteins, each contains a single target B-cell epitope (betaE) of beta-hCG. Two sets of DNA fragments were chemically synthesized encoding the beta5, beta9 and beta8 epitopes (betaE) 45 approximately 52, 113 approximately 116 or 133 approximately 144 of beta-hCG subunit and were inserted into the downstream of streptavidin (Stv) gene in pTSA18 separately, with or without an extra TAA codon at the 3'-terminals of the genes. SDS-PAGE analysis revealed that only Stv-betaE (-beta5, -beta9 or -beta8) fusion genes set with the TAA codon can be expressed in E. coli BL21 (DE3) pLysS strain at high level after 1mM IPTG induction for 4 hours. Additionally, these fusion proteins can all be recognized by specific polyclonal antiserum (RS-4157) generated upon immunization with the loop peptide 38 approximately 57 of beta-hCG, monoclonal antibody (mAb) FB12 to beta9 epitope and mAb OT3A that specially recognizes reporter sequence 133 approximately 139 of beta8 epitope 137 approximately 144. Each of the proteins can be purified to 95% relative homogeneity using an improved method of preparative gel polyacrylamide gel electrophoresis. The yields were 5 mg per 1 L culture. The three target Stv-betaE fusion proteins will be useful in determining the immunogenicity of designed hCG CPs and hCG vaccines, including hCG DNA vaccines.

Effects of TNF-alpha receptor blocking peptide on adjuvant arthritis in rats / 药学学报

<p><b>AIM</b>To study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.</p><p><b>METHODS</b>The model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.</p><p><b>RESULTS</b>The model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.</p><p><b>CONCLUSION</b>The TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.</p>