Wild-type p53-induced Phosphatase 1 Deficiency Exacerbates Myocardial Infarction-induced Ischemic Injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Myocardial infarction (MI) is a major disease burden. Wild-type p53-induced phosphatase 1 (Wip1) has been studied extensively in the context of cancer and the regulation of different types of stem cells, but the role of Wip1 in cardiac adaptation to MI is unknown. We investigated the significance of Wip1 in a mouse model of MI.</p><p><b>METHODS</b>The study began in June 2014 and was completed in July 2016. We compared Wip1-knockout (Wip1-KO) mice and wild-type (WT) mice to determine changes in cardiac function and survival in response to MI. The heart weight/body weight (HW/BW) ratio and cardiac function were measured before MI. Mouse MI was established by ligating the left anterior descending (LAD) coronary artery under 1.5% isoflurane anesthesia. After MI, survival of the mice was observed for 4 weeks. Cardiac function was examined by echocardiography. The HW/BW ratio was analyzed, and cardiac hypertrophy was measured by wheat germ agglutinin staining. Hematoxylin and eosin (H&E) staining was used to determine the infarct size. Gene expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) was assessed by quantitative real-time polymerase chain reaction (qPCR), and the levels of signal transducers and activators of transcription 3 (stat3) and phosphor-stat3 (p-stat3) were also analyzed by Western blotting. Kaplan-Meier survival analysis, log-rank test, unpaired t-test, and one-way analysis of variance (ANOVA) were used for statistical analyses.</p><p><b>RESULTS</b>Wip1-KO mice had a marginally increased HW/BW ratio and slightly impaired cardiac function before LAD ligation. After MI, Wip1-deficient mice exhibited increased mortality (57.14% vs. 29.17%; n = 24 [WT], n = 35 [Wip1-KO], P< 0.05), increased cardiac hypertrophy (HW/BW ratio: 7 days: 7.25 ± 0.36 vs. 5.84 ± 0.18, n = 10, P< 0.01, and 4 weeks: 6.05 ± 0.17 vs. 5.87 ± 0.24, n = 10, P > 0.05; cross-sectional area: 7 days: 311.80 ± 8.29 vs. 268.90 ± 11.15, n = 6, P< 0.05, and 4 weeks: 308.80 ± 11.26 vs. 317.00 ± 13.55, n = 6, P > 0.05), and reduced cardiac function (ejection fraction: 7 days: 29.37 ± 1.38 vs. 34.72 ± 1.81, P< 0.05, and 4 weeks: 19.06 ± 2.07 vs. 26.37 ± 2.95, P< 0.05; fractional shortening: 7 days: 13.72 ± 0.71 vs. 16.50 ± 0.94, P< 0.05, and 4 weeks: 8.79 ± 1.00 vs. 12.48 ± 1.48, P< 0.05; n = 10 [WT], n = 15 [Wip1-KO]). H&E staining revealed a larger infarct size in Wip1-KO mice than in WT mice (34.79% ± 2.44% vs. 19.55% ± 1.48%, n = 6, P< 0.01). The expression of IL-6 and p-stat3 was downregulated in Wip1-KO mice (IL-6: 1.71 ± 0.27 vs. 4.46 ± 0.79, n = 6, P< 0.01; and p-stat3/stat3: 1.15 ± 0.15 vs. 1.97 ± 0.23, n = 6, P< 0.05).</p><p><b>CONCLUSION</b>The results suggest that Wip1 could protect the heart from MI-induced ischemic injury.</p>

Effects of simvastatin on the proliferation of HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of simvastatin on the proliferation, cell cycle and expression of cyclin-dependent kinase inhibitor p21 protein in human hepatocellular carcinoma (HepG2) cells in vitro.</p><p><b>METHODS</b>HepG2 cells were administrated with simvastatin. Proliferation of the cells was detected by MTT assay, cell cycle was measured by flowcytometry and the cyclin-dependent kinase inhibitor p21 protein expression was detected by immunocytochemistry. The results were evaluated by factorial design and one-way analysis of variance.</p><p><b>RESULTS</b>Simvastatin inhibited HepG2 cells growth in vitro (F(concentration) = 1264, P value less than 0.001; F(time) = 17.466, P value less than 0.001; F(concentration*time) = 35.053, P value less than 0.001) and could arrest HepG2 cells in G0/G1 phase of cell cycle. However, apoptosis of HepG2 cells was not obvious. Simvastatin could also increase cyclin-dependent kinase inhibitor p21 protein expression (F = 512.133, P value less than 0.001).</p><p><b>CONCLUSION</b>Simvastatin can inhibit the growth of HepG2 cells in vitro, which may be explained by its effects of enhancing cyclin-dependent kinase inhibitor p21 protein expression and arresting HepG2 cells at G0/G1 phase of cell cycle.</p>

Construction of cholecystokinin transgenic mouse and its effects on food intake / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cholecystokinin (CCK) is one of the richest neuropeptides in the mammalian brain, which is mainly distributed in the cerebral cortex, hippocampus, thalamus and caudate-putamen. CCK is implicated in a variety of behavioral functions such as food intake, learning, memory, anxiety, pain and neuroprotection. The current research results for CCK are obtained mainly through injection of CCK peptide into the body. The key issues of whether CCK can regulate diet by a central pathway and whether there are long-term regulation effects on diet are still unresolved. In this study, the effects of CCK on food intake in transgenic mice were investigated.</p><p><b>METHODS</b>Transgenic mice were created by microinjection of the PDGF-CCK construct into male pronucleus of the zygotes. The genomic phonetype of transgenic mice were identified by PCR. The expression of PDGF-CCK was analyzed by Western blotting. Body weight, plasma glucose, cholesterol and triglycerides were assayed and analyzed.</p><p><b>RESULTS</b>Two PDGF-CCK transgenic independent lines were established and exhibited a high-levels brain-specific transgene expression compared with that of nontransgenic littermate controls. The food intake of male CCK transgenic mice was decreased by 5% - 10% with the same levels of water consumed compared with wild type mice. The food intake in female mice was not obviously changed. In the transgenic mice the bodyweight was lower and plasma glucose was higher compared with the nontransgenic littermate controls.</p><p><b>CONCLUSIONS</b>The high expression of the CCK gene in the brain can decrease body weight and increase plasma glucose. The differences in food intake between the males and females require further study.</p>

Screening and cloning gene of a hepatocyte protein interacting with HCV NS5ATP4A protein / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen and clone hepatocyte protein interacting with hepatitis C virus NS5ATP4A protein for studying its biological functions.</p><p><b>METHODS</b>Bait plasmids of hepatitis C virus NS5ATP4A were constructed. After verifying that hepatitis C virus NS5ATP4A protein could be steadily expressed in AH109 yeast strain, yeast-two hybrid assay was performed by mating AH109 with Y187 which pre-transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X-a-gal activity. Nineteen yeast colonies which grew on QDO and had a-gal activity were obtained, and then the library plasmids were extracted and sequenced.</p><p><b>RESULTS</b>Seven genes were screened out and one of them was a formerly unknown gene. They were associated with RNA synthesis, protein translation, cell cycling and tumor immunity.</p><p><b>CONCLUSION</b>NS5ATP4A binding proteins were successfully screened, which offers new clues for further studying the signal transduction pathway of NS5ATP4A and the pathogenic mechanism of HCV.</p>

Effects of human alpha-mannosidase Man2c1 transgene on growth and metastasis of transplanted tumor in mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effect of human alpha-mannosidase Man2c1 transgene on tumor growth and metastasis in mice.</p><p><b>METHODS</b>Hepatoma cell H22 or squamous epithelial carcinoma cell S180 was subcutaneously inoculated into the right armpit of mice (wild type mice and 28#, 35#, and 54# transgenic mice). Tumor size was measured every week. Mice were sacrificed on day 9 or 10 and then the tumors were exercised and weighted. Tumors and lungs were fixed in formaldehyde and sectioned. The sections were stained with hematoxylin/eosin and examined under microscope. The red blood cells in spleen were destroyed by Tris-NH4Cl. Natural killer (NK) cell activity was detected with Yac-1 cell as target.</p><p><b>RESULTS</b>H22 and S180 tumors grew faster in all the three transgenic mice (28#, 35#, and 54#) than in wild type mice. The average size and weight of tumors between the transgenic mice and wild type mice were significantly different (P<0.05). Most tumors in the transgenic mice invaded the surrounding tissues. In contrast, nearly all the tumors in wild type mice were capsulized. Three of 10 28# transgenic mice, 5 of 10 35# transgenic mice, 3 of 10 54# transgenic mice, and 1 of 10 wild type mice showed lung metastasis of H22 tumor. Two of 6 28# transgenic mice, 3 of 6 35# transgenic mice, 1 of 6 54# transgenic mice, and 0 of 6 wild type mice showed lung metastasis of S180 tumor. No difference of NK activity in spleen cells was observed between the transgenic mice and wild type mice.</p><p><b>CONCLUSIONS</b>hMan2c1 transgene promotes growth, invasion, and metastasis of transplanted H22 and S180 tumors in mice. hMan2cl transgene does not affect NK activity in splenocytes.</p>

Secretory Expression of the Fusion Protein IFN?-HSA in Pichia pastoris / 中国生物工程杂志

Overlapping PCR technology was employed to splice IFN?and HSA genes in vitro. The spliced gene was inserted into Pichia pastoris secretory vector pPIC9K. The IFN?-HSA gene was designed for secretory expression under the control of promoter AOX1 and Mat a signal peptide in pPIC9K. The recombinant plasmid pPIC9K/IFN?-HSA was linearized by restriction enzyme SalI and transformed into Pichia pastoris KM71 by electroporation. The recombinant strains identified by G418 selection and confirmed by PCR analysis were induced by methanol to express fusion protein IFNp-HSA. SDS-PAGE and Western blot analysis of the fusion protein showed that the expressed fusion protein IFNp-HSA with an apparent 90kDa molecular weight had the antigenicity of HSA. The specific activity of culture supernatant was about 640IU/ml assayed by the standard amiviral activity test on WISH cells challenged with VSV virus.

Cyclosporin A treatment of 83 children with nephrotic syndrome of different pathological types / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of cyclosporin A (CyA) therapy in 83 children with nephrotic syndrome of different pathological types.</p><p><b>METHODS</b>Eighty-three children enrolled in this study were all hospitalized children with idiopathic nephrotic syndrome, aged 3 to 14 yrs (average 8.3 yrs) and included 52 males and 31 females. There were 35 cases with steroid-dependent, 17 with steroid resistant and 26 with frequent relapses. CyA was given to each patient with dosage of 5 mg/(kg.d) during the corticosteroid was diminished. The renwal biopsy was performed in all patients before the administration of CyA. The duration of CyA therapy lasted for about 3 to 6 months. The plasma concentration of CyA was monitored.</p><p><b>RESULTS</b>Eighty-three children with nephrotic syndrome of different pathological types were treated with CyA, including 42 cases of minimal change nephrotic syndrome (MCNS), 31 cases of mesangioproliferative glomerulonephritis (MsPGN), 5 cases of membranoproliferative glomerulonephritis (MPGN) and 4 cases of focal segmental glomerular sclerosis (FSGS). All the 83 patients tolerated well to the CyA treatment. Forty-five cases got complete remission, 23 partial remission, 15 cases no change after one month treatment with CyA in the hospital. The overall response rate was 82%. Patients with different renal pathological types showed different responses. Among them, MCNS and MsPGN exhibited the best response rates of 86% and 84%, respectively; MPGN cases showed a lower response rate and FSGS cases showed the lowest rate. The response time was 7 to 45 days. The blood concentration of CyA was monitored for 1 week and 2 weeks after the drug was given. The effective drug concentration was maintained at 100 to 200 microg/L, and the course lasted for 3 to 6 months. During the follow-up of 83 cases, in 17 of 68 cases the disease relapsed when therapy was tapered or discontinued. The relapse rate was 25%. The results indicated that CyA would be effective to the relapsed cases. The serum creatinine increased temporarily after administration of CyA in 5 cases, N-acetyl-beta-D-glucosaminidase (NAG) in 8 cases and eventually reached the normal range after the adjustment of dosage. The side effects included anorexia, nausea, vomiting and so on.</p><p><b>CONCLUSION</b>CyA is one of the effective substitutes for the treatment of nephrotic syndrome, especially for the cases with MCNS and MsPGN. And CyA could control refractory nephrotic syndrome effectively and rapidly. The clinical effect was related to the blood concentration of CyA and pathological types.</p>

Effects of lidocaine on adhesion molecules expression in lung injury after ischemic shock in rats / 中华麻醉学杂志

Objective To study the effect of lidocaine on the expression of CD11b/CD18 and MPO in lung injury after hemorrhagic shock in rats. Methods Eighty male Wistar rats weighing 230-270 g were randomly divided into 4 groups: sham operation group ( group Ⅰ, n = 8 ) received operation without shock, shock group ( group Ⅱ, n = 8 ) received hemorrhagic shock without resuscitation , normal saline treatment group (group Ⅲ, n = 32 ) received normal saline after shock, lidocaine treatment group ( group Ⅳ, n = 32 ) received lidocaine after shock. The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg? kg-1 . Hemorrhagic shock was induced by withdrawing blood from right femoral artery at the rate of 2.0 ml?kg-1? min-1 , MAP was maintained at 40 mm Hg for 60 min before resuscitation. Direct arterial MAP and HR were continuously monitored. Group Ⅳwas received lidoacaine at the beginning of resuscitation with a bonus dose 2.0 mg? kg-1 and then followed continuous infusion1.0mg?kg-1?h-1 for 2 h. Group III was received the same dose of normal saline. Flow cytometric analysis was used to assess the expression of CD11b/CD18 on leukocyte and the change of myeloperoxidase (MPO) in the lung was studied at the end of shock (groupⅡ ) and at 2,4, 8, 12 h after resuscitation (group Ⅲ, group Ⅳ) . The rats were sacrificed and the piece of lung was immediately removed for electron microscopic examination. Result In group Ⅲand group Ⅳ ,the expression of CD11b/CD18 on PMNs and MPO in lung were increased significantly compared with Group Ⅰ. Microscopic examination indicated the longer time of reperfusion ,the more serious injury of the lung. And in groupⅣ ,the changes of CD11b/CD18 and MPO were reduced as compared with group Ⅲ (P