SUMO specific peptidase 3 regulates autophagy in mouse testicular Sertoli cells / 上海交通大学学报（医学版）

Objective: To investigate the regulation of autophagy by SUMO specific peptidase 3 (SENP3, normally called SUMO specific protease 3) in mouse testis. Methods: Immunofluorescence was used to detect the localization of SENP3 in spermatogenic cells and Sertoli cells of testis. Senp3 wild type (Senp3+/+) mice and Senp3 gene knockout heterozygous (Senp3+/-) mice were subjected to starvation treatment to induce autophagy. Testicular tissue proteins were extracted, and the extent of autophagy was detected by Western blotting. The extent of autophagy of Sertoli cells was detected and compared with that of spermatogenic cells in testis by transmission electron microscopy and immunofluorescence. Results: SENP3 mainly localized in the nucleus of Sertoli cells. Compared to Senp3+/+ mice, the extent of starvation-induced autophagy in Sertoli cells of Senp3+/- mice increased. Conclusion: SENP3 can inhibit autophagy in Sertoli cells during nutrient deficiency, which may play a role in controlling the extent of autophagy.

Identification and Characterization of Alternaria iridiaustralis Causing Leaf Spot on Iris ensata in China

In 2016, a severe leaf spot disease was found on Iris ensata Thumb. in Nanjing, China. The symptom was elliptical, fusiform, or irregularly necrotic lesion surrounded by a yellow halo, from which a small-spored Alternaria species was isolated. The fungus was identified as Alternaria iridiaustralis based on morphological characteristics. The pathogenicity tests revealed that the fungus was the causal pathogen of the disease. Phylogenic analyses using sequences of ITS, gpd, endoPG, and RPB2 genes confirmed the morphological identification. This study is the first report of A. iridiaustralis causing leaf spots on I. ensata in China.

Chinese Pediatric Reference Intervals for Serum Cortisol on IMMULITE 2000

Clinical interpretation of the test results for cortisol based on continuous reference intervals with appropriate partitions improves pediatric diagnosis; however, these values are available only for Caucasians. To develop the pediatric reference intervals for Chinese population, we examined the serum cortisol levels in 1,143 healthy Chinese children aged 4–18 years (566 boys and 577 girls), using an IMMULITE 2000 Immunoassay System (Siemens Healthcare GmbH). Phlebotomy was performed at 7–9 a.m. for 284 boys and 287 girls and at 1–3 p.m. for the others. They were divided into four age groups according to the Clinical and Laboratory Standards Institute guideline EP28-A3c, with the last group further stratified according to sampling time. Separate reference intervals of 49.6–323.7, 70.9–395.3, and 90.1–448.7 nmol/L were established for children aged 4–8, 9–12, and 13–15 years, respectively. Further, reference intervals of 118.2–464.7 and 71.4–446.7 nmol/L were established for morning and afternoon cortisol levels, respectively, in children aged 16–18 years. Further studies are necessary to transfer and validate these reference intervals in other analytical systems and pediatric populations, and to allow for broader applications.

Relationship between hepatitis B virus DNA and hepatitis B virus large envelope protein / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the relationship between HBV DNA and hepatitis B virus large envelope protein (HBV-LP) in patients with chronic hepatitis B.</p><p><b>METHODS</b>Serum HBV DNA was detected by RT-PCR and the HBV-LP was detected by enzyme linked immunosorbent assay in 320 serum samples collected from patients with chronic hepatitis B.</p><p><b>RESULTS</b>There were no significant difference between positive rate of HBV-LP and that of HBV DNA in different HBeAg patterns (P>0.05). Serum HBV-LP levels were closely correlated with HBV DNA copies (r=0.949).</p><p><b>CONCLUSION</b>Serum HBV-LP is a reliable serological marker that can reflect the replication of HBV.</p>

Study on the association of the gene polymorphism of thrombin activatable fibrinolysis inhibitor with coronary heart disease in Han population / 中华检验医学杂志

0.05)in the frequency of alleles and genotypes between controls and coronary heart disease.In additional,at the 325 position,the TAFI antigen of the Thr325Thr was higher[(114.89?2.53)%]than that of the other genotype(Thr325Ile and Ile325Ile),there was significant difference between the TAFI antigen of the Thr325Thr and the others(P 0.05).But the TAFI activity of the Ile325Ile was lower(3.08?3.63 ?g/ml)than that of the other genotypes(Thr325Ile and Thr325Thr),there was significantly difference between the TAFI activity of the Thr325Thr and the other(P

IL-3 Gene-Modified Bone Marrow Stromal Cells Accelerate Hematopoiesis Recovery of Mice Received High Dose Chemotherapy / 中国肿瘤生物治疗杂志

A main complication of chemotherapy in cancer patients is hematopoiesis suppression. Microenviroment transplantation using bone marrow stromal cells (BMSCs) has been demonstrated to be a potent method in recovery of hematopoiesis in animal models. Based on hematopoiesis-supportive ability of BMSCs and high potency of IL-3 in hematopoiesis stimulation, BMSCs were studied as a cellular delivery system for IL-3 gene transfection to promote hematopoiesis recovery of mice after high dose chemotherapy. BMSCs were transfected with recombinant adenovirous containing murine IL-3 gene(MOI = 10), the level of mIL-3 secreted by gene-modified BMSCs was 110U/ml/10~6 cells/ 24h in vitro. The mice were injected with high dose cyclophosphamide(200mg/kg) i.p. and after 24 hours the IL-3 gene-modified BMSCs(2 x 10~6/mouse) were transplanted intrasplenically. White blood cell counts in peripheral blood of mice received intrasplenic injection of IL-3-BMSCs were kept at a high level within two weeks after chemotherapy. The pathological sections of spleens and bone marrow showed significant recovery of hematopoiesis, compared with that of mice received chemotherapy only. The data indicated the feasibility of IL-3 gene-modified BMSCs transplantation in the acceleration of hematopoiesis recovery after chemotherapy.

Combined Use of CD Suicide Gene and Lymphotactin Gene Therapy Elicit Potent Antitumor Effects / 中国肿瘤生物治疗杂志

Adenovirus harboring E. coli. cytosine deaminase gene (AdCD) and adenovirus encoding with lymphotactin gene (AdLtn) were used for gene therapy in vivo. BALB/c mice were inoculated subcutaneously with CT26 colon adeno-carcinoma cells and 3 days later received combined injection of AdCD and/or AdLtn followed by continuous injection with 5-fluorocytosine(5-FC) 300mg/kg. The results demonstrated that mice received combined therapy developed tumors most slowly and survived longest when compared with mice treated with AdCD/5-FC, AdLtn, AdlacZ/5-FC or PBS. To further explain the immunological mechanism of the antitumor effects by the combined therapy, we found that combined treatment with suicide gene and Ltn gene therapy achieved maximal cytotoxic effects of nature killer cells and specific cy-totoxic T lymphocytes. FACS analysis of the tumor mass demonstrated that AdCD/5-FC in combination with AdLtn therapy increased the expression of H-2K~d and B7-1 expression on tumor cells. The CD4~+ and CD8~+ cells infiltrated in the tumor mass after combined therapy were significantly increased when measured by FACS analysis. Our results demonstrated that combined transfer of suicide gene and lymphotactin gene induce nonspecific and specific antitumor immunity of the host and elicit more potent antitumor effect.

More Efficieat Induction of Autitumor Response by Combined Suicide Gene and Interleukin 2 Gene Transfer Mediated by Recombinant Aden-ovirus / 中国肿瘤生物治疗杂志

Antitumor effect of combined transfer of suicide gene and cytokine gene was evaluated in the present study. Adenoviruses expressing E. coli. cytosine deaminase (AdCD) and adenoviruses expressing murine interleukin 2 (AdTL2) were used for the treatment of tumor-bearing mice. The mice were inoculated s. c. with FBL-3 leukemia cells and 3 days later received intratumoral injection of AdCD in the presence or absence of AdIL2 followed by intraperitoneal 5-fluorocytosine (5FC) administration. The results demonstrated that tumor-bearing mice treated with AdCD/5FC in combination with AdTL2 showed more .potent inhibition of tumor growth and survived much longer as compared with mice treated with AdCD/5FC, AdEL2, AdlacZ/5FC or PBS. It was illustrated that the tumor mass showed obvious necrosis and inflammatory cell infiltration, and more CD4+ and CD8+ T cells infiltrated into the tumor after combined therapy. The splenic NK and CTL activities increased significantly in mice after combined transfer of CD gene and EH gene. Our results demonstrated that combined transfer of suicide gene and IL-2 gene could inhibit the growth of established tumor in mice significantly and induce antitumor immunity of the host efficiently.

Induction of Tumor Cell Apoptosis by Adenovirus-Mediated E. coli Cyto-sine Deaminase Gene Transfection and Its Bystander eftect / 中国肿瘤生物治疗杂志

Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-

hG-CSF cDNA Cloning and the Construction and Applicaltion in Gene Therapy of Its Retroviral Vector / 中国肿瘤生物治疗杂志

The hG -CSF cDNA was cloned by RT-PCR and confirmed by sequencing, which contains the full length of hG-CSF encoding region and parts of 5 '、3' non - coding region. Then the hG - CSF retroviral vector pLGSN was constructed by orientationally inserting the hG-CSF cDNA into the EcoRI/XhoI cloning sites of pLXSN vector. Packaged with CRE and CRIP packaging cell lines which are considered to be unlikely to produce helper viruses, the final pLGSN retrovirion titer reached 1. 1 ?106 CFU/ml. During constitutive passaging, the CRIP - LGSN cell clone produced relatively stable tilers of pLGSN retrovirion, ranging from 6. 8?105CFU/ml to 1. 1?106CFU/ml. By infecting the murine fibroblast cell line NIH3T3 with pLGSN retrovirion, a cell clone designated as NIH3T3 -G -CSF was obstained, secreting 168U/ml G-CSF . The integration and expression of hG-CSF gene in this cell clone were confirmed by Southern and Northern blotting analyses. Western blotting has also detected specifically the hG-CSF protein in the condensed supernalants from NIH3T3-G-CSF cells . After packaging the hG-CSF-secreting fibroblasts with collagen and implanting them into synergenic mice peritoneally , we detected a certain levels of G-CSF in the sera of mice, which suggested the implanted NIH3T3-G-CSF fibroblast cells could constitutively express and release hG-CSF in vivo. Our data showed the constructed hG-CSF retroviral vector could be used to further investigate the fibroblasl-mediated hG-CSF gene therapy .