Development and verification of whole column imaging detection-capillary isoelectric focusing method for determination of isoelectric point of pertactin / 中国生物制品学杂志

@#Objective To develop and verify a whole column imaging detection-capillary isoelectric focusing(WCID-CIEF)method for the determination of isoelectric point(pI)of pertactin(PRN).Methods The WCID-CIEF method for the determination of PRN antigen was developed by optimizing the parameters such as the focusing time and final concentration of samples in the WCID-CIEF process,and verified for the specificity,accuracy,repeatability,intermediate precision,durability and inter-batch consistency.Results The optimal focusing time of WCID-CIEF for the determination of PRN antigen pI was 1 min at 1 500 V and 3 min at 3 000 V. The optimal final concentration of PRN antigen was 300 μg/mL. The PRN antigen pI was about 6. 035,and the blank matrix showed no interference peak in the position of each peak of antigen. The method had good specificity,accuracy,repeatability,intermediate precision,durability and consistency among batches.Conclusion The developed WCID-CIEF method is suitable for the pI detection and charge heterogeneity analysis of PRN antigen,which can provide basis for the characterization of PRN antigen and reference for the quality control in the process of development and production of related vaccines.

Gene mutation profiling and clinical significances in patients with renal cell carcinoma

Abstract Objectives The pathological mechanisms of patients with Renal Cell Carcinoma (RCC) remain defined. This study aimed to evaluate relationships between the landscape of gene mutations and their clinical significance in RCC patients. Methods Tissue and peripheral blood samples of 42 patients with RCC were collected and performed for the Next Generation Sequencing (NGS) with Geneseeq PrimeTM 425-gene panel probes. Their landscapes of gene mutation were analyzed. We also carried out an evaluation of Tumor-Node-Metastasis (TNM) staging, RENAL nephelometry score, surgery, and targeted drug treatment of patients. Then we compared the correlations of landscape in gene mutations and the prognosis. Results The most common gene alternations, including BAP1, PBRM1, SETD2, CSF1R, NPM1, EGFR, POLE, RB1, and VHL genes, were identified in tissue and blood samples of 75% of patients. EGFR, POLE, and RB1 gene mutations frequently occurred in relapsed and metastatic patients. BAP1, CCND2, KRAS, PTPN11, ERBB2/3, JAK2, and POLE were presented in the patients with > 9 RENAL nephelometry score. Univariable analysis indicated that SETD2, BAP1, and PBRM1 genes were key factors for Disease-Free Survival (DFS). Multivariable analysis confirmed that mutated SETD1, NPM1, and CSF1R were critical factors for the Progression Free Survival (PFS) of RCC patients with target therapy. Conclusions Wild-type PBRM1 and mutated BAP1 in patients with RCC were strongly associated with the outcomes of the patient. The PFS of the patients with SETD2, NPM1, and CSF1R mutations were significantly shorter than those patients without variants.

Paeoniflorin-6'-O-benzene sulfonate ameliorates progression of adjuvant-induced arthritis by inhibiting interaction between Ahr and GRK2 of fibroblast-like synoviocytes / 中国药理学与毒理学杂志

OBJECTIVE Aryl hydrocarbon receptor (Ahr) is thought to be a crucial factor that regulates immune responses, which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis (RA). The results of our group in recent years have shown that CP-25, a novel ester derivative of paeoniflorin, has a good effect on improving RA animal models. However, whether the anti-arthritis effect of CP-25 is related to Ahr remains unclear. METHODS CP-25 treatment ameliorated adjuvant-induced arthritis (AA), a mouse model of RA, by inhibiting Ahr-related activities in fibroblasts like synoviocytes (FLS). AA rats were treated with CP-25 or paroxetine from day 17 to 33 after immunization. RESULTS CP-25 alleviated arthritis symptoms and the pathological changes, decreased the expression of Ahr in the synovium and FLS of AA rats. Besides, treatment with CP-25 reduced the proliferation and migration of MH7A caused by Ahr activation. In addition, we also demonstrated that CP-25 down-regulated the co-expres?sion and co-localization of Ahr and G protein-coupled receptor kinase 2 (GRK2) in MH7A. CONCLUSION The data pre?sented here demonstrated that CP-25 suppressed FLS dysfunction in rats with AA, which were associated with reduced Ahr activation and the interaction between Ahr and GRK2.

Expression of lncRNA PVT1 in colorectal cancer tissues and cells and its effect on chemo-sensitivity to cisplatin and the possible mechanisms / 中国肿瘤生物治疗杂志

@#Objective: To study the regulatory effects and possible mechanism of long non-coding RNA plasmacytoma variant translocation 1 (lncRNA PVT1) on chemotherapy sensitivity to cisplatin (DDP) of colorectal cancer (CRC)．Methods: A total of 112 pairs of matched cancer and adjacent non-cancerous tissues were obtained from the CRC patients who underwent surgical resection in the First Affiliated Hospital of Xinxiang Medical University betweenApril 2006 and March 2011.All specimens were confirmed by pathological examinations. Tumor tissues and corresponding adjacent non-cancerous tissues from 30 cisplatin-sensitive CRC patients and 30 cisplatin-resistant patients were selected. Human CRC cell lines (HT29, SW480, HCT116, RKO and LoVo) and normal colonic epithelial cell line NCM460 were also collected for this study; and DDP-resistant RKO/DDP and LoVo/DDP cell lines were constructed. siPVT1, siNC, LV-PVT1 and LV-NC were transfected into LoVo and RKO cells or LoVo/DDP and RKO/DDP cells using lipofectamineTM2000. The expression of lncRNA PVT1 in CRC tissues and cells was tested by Real-time qPCR. CCK-8 assay, flow cytometry and WB were performed to test the effect of PTV1 knockout or enforcement on cell proliferation, apoptosis and expressions of apoptosis-related proteins, respectively. The CRC subcutaneous transplanted xenograft model was established on athymic nude mice to study the effect of PVT1 over-expression on tumor growth and DDP resistance. Results: PVT1 was highly expressed in the cancer tissues and CRC cells, and its expression was positively associated with cisplatin resistance of CRC. After knockdown of PVT1, the proliferation of cisplatinresistant CRC cells was significantly suppressed, while the apoptosis was significantly enhanced (P<0.05 or P<0.01); Mechanically, the levels of drug resistance-associated molecules, including MDR1 and MRP1, as well as the expression of anti-apoptotic Bcl-2 were significantly downregulated whereas the levels of pro-apoptotic Bax and cleaved caspase-3 were increased in PVT1-silenced DDP-resistant CRC cells. Over-expression of PVT1 reversely increased proliferation and decreased apoptosis of CRC cells (P<0.05 or P<0.01). In addition, PVT1 over-expression in CRC cells significantly promoted DDP-resistance in vivo (P<0.05). Conclusion: Collectively, knockdown of PVT1 expression can significantly suppress cell proliferation and promote apoptosis of DDP-resistant CRC cells. Overexpression of PVT1 can significantly promote the growth of CRC cells in vitro and transplanted xenograft in vivo. PVT1 regulates endogenous apoptosis pathways and further promotes the sensitivity of CRC cells to cisplatin chemotherapy via inhibiting the expressions of MDR1 and MRP1.

Suicidal status and its risk factors of people living with HIV/AIDs / 上海交通大学学报（医学版）

Although antiretroviral therapy appears to prolong the lifespan of people living with HIV/AIDS (PLWH), but the suicidal risk of these people is still significantly higher than the general population. Some PLWH had suicidal ideation, attempted suicide, and suicidal plan. The reasons for the high risk among these people mainly include as follows. ①The demographic factors include gender, sexual orientation, age, religion, race, etc. ②The social psychological factors are stressors (stressful or traumatic life events, social or interpersonal problems and mental illness), stigma, hiding HIV history, etc. Social or interpersonal problems include discrimination, social isolation, lack of social support, etc. Mental diseases include depression, anxiety disorders, substance use disorders, etc. In order to reduce the suicidal risk of PLWH, the government and related organizations can research and improve these social psychological factors.

Familial progressive hyper- and hypopigmentation: a report on a Chinese family and evidence for genetic heterogeneity

Abstract Background: Familial progressive hyper- and hypopigmentation (FPHH) is a rare genodermatosis that is characterized by diffuse hyper- and hypopigmented spots on the skin and mucous membranes. It is caused by a pathogenic mutation of the KITLG gene. Objectives: To investigate the clinical features and mutation of the KITLG gene in a Chinese family with FPHH. Methods: Histopathological and immunohistochemical analysis of lesions from the proband was performed. The KITLG gene was screened for the presence of mutations. Results: A Chinese family containing 14 individuals with FPHH was described, and the proband was a 5-year-old girl showing diffuse hyper- and hypopigmented lesions on her extremities and trunk. Histopathological and immunohistochemical staining for S100 and HMB45 of skin biopsy specimens from the hyperpigmented areas showed a striking increase in melanin throughout the epidermis, especially in the basal cell layer, and staining of hypopigmented area specimens displayed lower levels of melanin in the epidermis. Mutation analysis of the KITLG gene was performed, but no mutation was found. Study limitations: The new pathogenic gene was not found. Conclusion: A family with FPHH was described. Analysis revealed that its members did not have any mutations of the KITLG gene, which provided evidence for genetic heterogeneity of this genodermatosis.

Effect of Qibai Pingfei capsule medicated serum on protein expressions of KATP channel in pulmonary arterial smooth muscle cells via nitric oxide / 中国中药杂志

To investigate the ATP-sensitive potassium channel (KATP channel) protein expressions during different periods under hypoxia condition and explore the effect of Qibai Pingfei capsule medicated serum (hereinafter referred to as QBPF) on the correlation between the protein expressions of KATP channel and nitric oxide in rat pulmonary arterial smooth muscle cells(PASMCs). Qibai Pingfei capsules were given to SD rats via continuous gavage for 10 days to obtain QBPF. Primary rats PASMCs were cultured by the direct adherent culture method. Western blot was applied to detect the protein expression levels of KATP channel (Kir6.1 and SUR2B) in PASMCs. Then the noncompetitive inhibitor of NO synthase--Nω-nitro-L-arginine methyl ester(L-NAME) and KATP channel inhibitor--glyburide(GLYB) were applied respectively to evaluate the effect of QBPF on the protein expressions of KATP channel. The protein expressions of Kir6.1 and SUR2B were increased after 6-hour hypoxia treament, peaked at the 24-hour hypoxia treament, and decreased in both 48-hour and 72-hour hypoxia groups. Especially, QBPF could further up-regulate the Kir6.1 and SUR2B protein expressions under 24-hour hypoxia condition; however, such up-regulation effect could be blocked by KATP channel inhibitor GLYB and NO specific inhibitor L-NAME, indicating that QBPF played the role of opening KATP channel. The regulatory mechanism was probably associated with up-regulating KATP channel protein expression via NO relative pathway, involving pulmonary vasodilation, and thus relieving the occurence and development of COPD.

Cluster analysis of 2007 survey data of current status of the national brick-tea-born fluorosis / 中国地方病学杂志

ObjectiveCluster analysis method was applied to analyze the data of brick-tea-born fluorosis,to clarify the provincial and municipal classification hierarchy in nation-wide epidemic areas and to guide the prophylaxis and treatment of the disease.MethodsPrevalence survey database of the 2007 national brick-tea-born fluorosis was analyzed.Detection rate of second degree and above clinical skeletal fluorosis as clustered index,clustering analysis of all survey data was carried out,clustering figure was drawn,and analysis of variance and multiple comparison was done.ResultsThe provincial level of the nation-wide epidemic areas of the brick-teaborn fluorosis could be classified into three categories:the first category including Tibet and Sichuan province,the second category including Qinghai province,and the third including Inner Mongolia,Gansu,Xinjiang and Ningxia provinces; The city-level they were also could be classified into three categories:the first category including Aba,Shigatse region and Lhasa,the second including Wulanchabu,Hulun Buir,Baotou,Qinghai state,Guoluo state,Haixi state,Xining city,Haidong state,Ganzi state,Longnan city,Jiuquan city,Urumqi city and the state of Bortala,and the other cities were the third category cities.The differences of clinical skeletal fluorosis detection rate between the first,second,and third provincial and municipal categories(35.18％,16.21％,5.22％,41.16％,19.64％,4.19％) were statistically significant (all P ＜ 0.05).ConclusionsBrick-tea-born fluorosis can be classified by using cluster analysis at the provincial and municipal levels.Cluster analysis results can provide a scientific basis for effective implementation of the prophylaxis and treatment of brick-tea-born fluorosis.

Research progress on olfactory receptor / 浙江大学学报·医学版

The olfactory perception is the process that the olfactory receptor is activated by odorous molecules, which induce the transduction of signal in the cell and the chemical information is transduced into electrical impulses. After the changed signal is transmitted to the brain, the whole perception process completes. OR gene belongs to the multigene family. The coded olfactory receptor proteins belong to the G-protein-coupled receptor (GPCR) superfamily and therefore are invariably seven-transmembrane domain(7TM) protein. Olfactory receptor protein plays an important role in olfactory perception and signal transduction process.

Clinical application of anatomy of blood vessel arches in ileum: Report of 45 cases / 中国医师杂志

ObjectiveTo observe anatomy of the blood vessel arches and their blood circulation in the ileum and further establish surgical strategy for preventing excessive tonicity and ischemia of the ileum after anastomosis of the ileum pouch and the anal canal.MethodsThe blood vessel arches were dissected in the ileum of 45 corpses and their blood circulation routes were observed.ResultsThe 2-tier blood vessel arches were found in 2 cases (4％), the 3-tier blood vessel arches in 35 cases (78％), the 4-tier blood vessel arches in 5 cases (11％), and the 5-tier blood vessel arches in 3 cases (7％).ConclusionsUnder the right colic artery, curing the superior mesenteric artery and vein can release the length of the small intestines, prevent tonicity and ischemia of the ileum after anastomosis of the ileum pouch and the anal canal, and supply enough blood to the ileum by the 2 -5 tier blood vessels.

Research on frequency of application with modern Chinese herbal medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the relationship between 500 kinds of commonly used Chinese herbal medicine and the classification of their efficacies in Chinese Materia Medica in relation to the common diseases listed in Internal Medicine.</p><p><b>METHODS</b>Database retrieval frequency of the quantitative statistical method was adopted. First, the 8 980 kinds of Chinese herbal medicine recorded in Chinese Materia Medica were used as the original search objects, and 4 493 kinds which were cited in more than five articles were picked out and then rechecked for further title citations. Second, as judged based on the Criterion, the numbers of articles which included the medicines in the line of standards were examined. As a result, 500 species of Chinese herbal medicine were singled out based on their retrieval frequency and were then used for compilation of the classification statistics according to their efficacy and the common diseases in Internal Medicine.</p><p><b>RESULTS</b>From the classification of Chinese medicines, herbs with wide efficiency and a meek nature had higher frequencies, but those which were not appropriate as decoctions had relatively lower frequencies. However, according to the average frequency, the Chinese herbal medicine for nourishing qi and tonifying blood, at 36,346 times and 34,544 times, respectively, were the most commonly used. Analyzed from the frequency of application of the Chinese medicine in the treatment of common diseases, most of the top 10 kinds of Chinese herbal medicine with the highest frequencies generally coincided with the 500 selected medicines. In addition, the Chinese medicines with clear pharmacological efficiency were easily isolated and purified to be made into injections, although other forms are more commonly used.</p><p><b>CONCLUSION</b>The results of the research objectively reflected the current applications of Chinese herbal medicine, and could be used as references in teaching, research, clinical applications, and in compiling and increasing the drugs in textbooks and Pharmacopoeia.</p>

Analysis on the present running status of water-improving project in Anda city,Heilongjiang province in 2008 / 中国地方病学杂志

Objective To understand the running status and distribution of water-improving pmjects in Anda city.Methods The running and management of the water-improving projects were investigated and located the position by Global Positioning Systerm(GPS)in Anda city.Results Among 317 water-improving projects,16.09%were either long-term projects or in poor management or had already stopped usage,77 projeets were broken,accounting for 24.29%;all inadequate supply of equipment and pipeline,83 projects had never been started,accounting for 26.18%;now,106 projects were running,accounting for 33.44%.In only 46 projects,the water fluoride concentration was lower than 1.00 mg/L,accounting for 14.51%,as a result 36 thousand people benefited.Conclusions The running status of water-improving projects was unacceptable,most of them stopped running and endemic fluorosis control was still severe in Anda city.

Effect of sustained release of recombinant rat insulin-like growth factor-1 from poly (lactide-CO-glycolide ) microspheres on bone formation in the peri-implant areas in Goto-Kakizaki rats with type 2 diabetes / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the effect of sustained release of recombinant rat insulin-like growth factor-1(rrIGF-1) from poly (lactide-CO-glycolide) (PLGA) microspheres on bone formation in the peri-implant areas in Goto-Kakizaki rats with type 2 diabetes.</p><p><b>METHODS</b>Type 2 diabetes models were successfully established in 20 male Goto-Kakizaki rats, which were then randomly divided into treatment group (sustained release of rrIGF-1 from PLGA microspheres were loaded on the peri-implant areas, n=10) and diabetic group (loaded with isodose placebo from PLGA microspheres, n=10). Another ten male SD rats served as control group (did not sustain any loading). Titanium implants were inserted into the tibias of 30 diabetic and normal animals. Four, 5, and 8 weeks after implantation, local blood samples around the implants were obtained for the determination of serum osteocalcin (OCN), serum bone specific alkaline phosphatase (B-ALP), and serum procollagen I carboxyterminal propeptide (PICP) with enzyme linked immunosorbent assays.</p><p><b>RESULTS</b>Four weeks after implantation, OCN, B-ALP, and PICP were significantly lower in both treatment group and diabetic group than in control group(both P<0.05). Five weeks after implantation, serum OCN and B-ALP levels of the diabetic group were significantly lower than those of the other two groups (all P<0.05). Serum PICP levels of both diabetic group and treatment group were significantly lower than that of control group(both P<0.05). The OCN level in the trealment group was significantly higher in the post-operative 5th week than in the post-operative 4th week, while the PICP levels in the diabetic group were significantly lower than those in the treatment group and control group in the post-operative 8th week (both P<0.05).</p><p><b>CONCLUSION</b>Sustained release of rrIGF-1 from PLGA microspheres loaded on the local peri-implant areas can promote bone formation in the peri-implant areas in Goto-Kakizaki rats with type 2 diabetes.</p>

Expression of Smad4 in prostate cancer LNCaP and ARCaP cell lines with different metastatic potentials and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the expression of Smad4, the core of TGF-beta/Smads signal transduction pathway in different prostate cancer cell lines, and explore their molecular mechanism of bone metastatic potential.</p><p><b>METHODS</b>The Millicell polycarbonate filter coated with matrigel was used to confirm the invasive potency of LNCaP and ARCaP cell lines (IF11 and IA8). The expressions of the Smad4 protein and mRNA in these prostate cancer cells with different metastatic potentials were detected by Western blotting and RT-PCR, respectively.</p><p><b>RESULTS</b>ARCaP cell lines (IF11 and IA8) exhibited a stronger potency of invasion than LNCaP (P < 0.01). The Smad4 protein and mRNA highly expressed in the LNCaP cell line that was well-known with a low metastatic potential, but not in the ARCaP (IF11 or IA8) cells with high metastatic potentials (P < 0.01).</p><p><b>CONCLUSION</b>Smad4 expresses differently in LNCaP and ARCaP cell lines with different metastatic potentials and, as a tumor suppressive gene, its deficient expression may play an important role in the invasion and metastasis of advanced prostate cancer.</p>

Transforming growth factor beta upregulates the expression of invasion and metastasis associated proteins in prostate cancer LNCaP cell lines in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the effect of the transforming growth factor beta (TGF-beta) on the expression of invasion and metastasis associated proteins in the prostate cancer LNCaP cell line in vitro.</p><p><b>METHODS</b>The prostate cancer cell line LNCaP was treated with TGF-beta in vitro. Western blotting was used to detect the expression of the "invasion and metastasis" associated proteins E-Cadherin, N-Cadherin and Vimentin.</p><p><b>RESULTS</b>The expression of N-Cadherin and Vimentin of the LNCaP cells treated with TGF-beta for 12 hours was significantly upregulated, but not that of E-Cadherin.</p><p><b>CONCLUSION</b>TGF-beta may induce epithelial-mesenchymal transition (EMT) of LNCaP cells which might be of importance in promoting prostate cancer cells invading to ambient tissues and metastasizing to distant organs.</p>

Over-expression of hypoxia-inducible factor 1 alpha increases angiogenesis of LNCaP cells in vivo / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the influence of hypoxia-inducible factor 1 alpha (HIF-1alpha) on angiogenesis in prostate carcinoma in vivo and to investigate its molecular mechanism.</p><p><b>METHODS</b>LNCaP/HIF-1alpha and LNCaP cells were cultured, the level of PSA in the supernatant of the culture medium detected by ELISA assay before and after the transfection, and the cellular cycle measured by flow cytometry. Nude mouse models of subcutaneous tumor were established with LNCaP/HIF-1alpha and LNCaP cells, the tumor growth observed, and tumor specimens collected for immunohistochemical staining.</p><p><b>RESULTS</b>Compared with the LNCaP cells, LNCaP/HIF-1alpha cells showed an obviously decreased PSA level (t = 8.243, P < 0.05) and enhanced proliferous activity. The tumorigenesis rate increased and the tumorigenesis time advanced in the LNCaP/HIF-1alpha group of the nude mice. Immunohistochemistry displayed higher expressions of VEGF, iNOS and Ang-2 in the LNCaP/HIF-1alpha than in the LNCaP group.</p><p><b>CONCLUSION</b>The over-expression of HIF-1alpha can up-regulate VEGF and iNOS involved in angiogenesis in vivo and contribute to the invasive potency of LNCaP cells. HIF-1alpha may have no influence on Ang-2 either in vitro or in vivo, while the expression of Ang-2 is regulated by other factors in vivo.</p>

Expressions of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma and their correlation / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the levels of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma tissues and explore their relationship.</p><p><b>METHODS</b>We enrolled in this study 42 patients with bladder transitional cell carcinoma, including Ta-T1 (n = 18), T2-T4 (n = 24), G1 (n = 12), G2 (n = 19), G3 (n = 11), metastasis (n =26) and non-metastasis (n = 16). Another 5 cases of normal bladder tissues were taken as controls, and the levels of MMP-2 and COX-2 mRNA were detected by RT-PCR.</p><p><b>RESULTS</b>The relative expressions of COX-2 mRNA were 1.038 +/- 0. 484 in Ta-T1, 1.489 +/- 0.584 in T2-T4, 0.920 +/- 0.442 in G1, 1.338 +/- 0.584 in G2 and 1.632 +/- 0.515 in G3, all significantly higher than that of the controls (0.460 +/- 0.224, P < 0.05). And the corresponding relative levels of MMP-2 mRNA were 1.107 +/- 0.384, 1.604 +/- 0.425, 0.971 +/- 0.370, 1.445 +/- 0.378 and 1.755 +/- 0.387, also significantly higher than that of the latter group (0.423 +/- 0.227, P < 0.05). The COX-2 and MMP-2 mRNA levels in the tumor tissues with and without metastasis were 1.591 +/- 0.455 vs 0.815 +/- 0.430 and 1.676 +/- 0.339 vs 0.927 +/- 0.228, (P < 0.01), respectively, with a positive correlation between the mRNA level of COX-2 and that of MMP-2 (r = 0. 703, P < 0.01).</p><p><b>CONCLUSION</b>MMP-2 and COX-2 mRNA are highly expressed in bladder transitional cell carcinoma tissues and their expressions are positively correlated with the degree of malignancy. MMP-2 and COX-2 might play a synergetic role in the pathogenesis and progression of bladder transitional cell carcinoma.</p>

Over-expression of hypoxia-inducible factor-1 alpha increases angiogenesis of LNCaP cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of hypoxia-inducible factor-1 alpha (HIF-la) on angiogenesis in human prostate cancer cells.</p><p><b>METHODS</b>Human prostate cancer cells of the line LNCaP were cultured and transfected by the recombinant plasmid pcDNA3. 1(-)/HIF-1alpha containing the gene HIF-1alpha with the Lipofectamine 2000 system. The positive clone cells were selected by G418 and confirmed by Western blotting and immunofluorescence (LNCaP/HIF-1alpha cells). The expressions of VEGF, iNOS and Ang-2 were detected by Western blotting.</p><p><b>RESULTS</b>The expression of HIF-1alpha in the LNCaP/HIF-1alpha cells was distinctly higher than that in the LN-CaP cells. Compared with the LNCaP cells, the expressions of VEGF and iNOS were up-regulated, whereas Ang-2 remained unchanged in the LNCaP/HIF-1alpha cells.</p><p><b>CONCLUSION</b>The over-expression of HIF-1alpha can induce an increase in angiogenesis proteins and improve the angiogenesis potency of prostate cancer.</p>

Apoptosis and expression of caspase-3 protein in germ cells of varicocele rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the relationship between the expression of caspase-3 in testicular germ cells of rats with experimental left varicocele (ELV) and apoptosis of germ cells.</p><p><b>METHODS</b>Twenty-four male SD rats were randomly divided into three groups with eight animals each: sham-operation group (SOG), 30-day post-operation group (PG1) and 60-day psot-operation group (PG2). ELV model was established by the partial ligation of the left renal vein. To detect apoptosis of germ cells and expression of caspase-3, TUNEL assay and immunohistochemistry (SABC) were used respectively.</p><p><b>RESULTS</b>The number of caspase-3 positive germ cells per tubular cross section in left and right testes of rats in SOG, PG1, PG2 were 0.1175 +/- 0.0129, 0.2463 +/- 0.0421, 0.2938 +/- 0.0511 and 0.1650 +/- 0.0192, 0.2538 +/- 0.0219, 0.2775 +/- 0.0343, respectively. Compared with SOG, the expression of caspase-3 in bilateral testes of rats in PG1 and PG2 were increased, and the differences were statistically significant(P = 0.0115 and P = 0.0144).</p><p><b>CONCLUSION</b>Expression of caspase-3 protein increased in germ cells of rats with ELV, which may be one of the molecular mechanisms related to excessive testicular germ cell apoptosis.</p>

The cloning, high level expression in Escherichia coli of human beta-defensin 3 and its antimicrobial activity analysis / 生物工程学报

In recent years, Bacterial resistance is more and more serious for the irrational use of antibiotics produces resistant strains and other reasons. Human are trying to solve the problem from different ways, including the study of antimicrobial peptides. Defensin is one of the most important of antimicrobial peptides. A novel antimicrobial peptide, human beta-defensin 3, was isolated and demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes. The total RNA was extracted from human tonsil and the hbetaD-3 specific cDNA sequence was amplified with RT-PCR. After sequenced, the target DNA fragment was cloned into pQE-80L vector together with the DNA fragment encoding carrier protein DHFR. The recombinant vectors were transformed into E. coli M15 and the expression was induced based on the optimal values of the IPTG concentration incubation temperature and induction time determined in the previous section. The expressed proteins were analyzed by SDS-PAGE and Western-blotting. The mass of the recombinant protein was about 40% of total bacteria protein. Isolate and purify the target protein. The recombinant hbetaD-3 fusion proteins possess the antimicrobial activity to staphylococcus aureus, multiresistant staphylococcus aureus (only vancomycin-sensitive) and Candida albicans in the assay of drug susceptibility. Advanced study can be continued based on our experiments.