Artificial neural network-genetic algorithm-based optimization of aerobic composting process parameters of Ganoderma lucidum residue.

The rapid development of traditional Chinese medicine enterprises has put forward higher requirements for the resource utilization of traditional Chinese medicine residues (TCMR). Aerobic composting of TCMR to prepare bio-organic fertilizer is an effective resource utilization method. In this study, a back-propagation artificial neural network (BPNN) model using composting factors as inputs (C/N, initial moisture content, type of inoculant, composting days) and the humic acid content as the output was constructed based on the orthogonal test data. BPNN-GA (a genetic algorithm) was used for extreme value optimization, and the optimal composting process parameter combination was obtained and verified. The results show that the combination of orthogonal testing and BPNN can effectively establish the relationship between the composting process parameters and humic acid content. The R2 value was 0. 9064. The optimized parameter combination is as follows: C/N,37.42; moisture content,69.76%; bacteria,no; and composting time,50 d.

Patients with SERPINC1 rs2227589 polymorphism found to have multiple cerebral venous sinus thromboses despite a normal antithrombin level: A case report.

BACKGROUND: The hereditary antithrombin (AT) deficiency caused by SERPINC1 gene mutation is an autosomal dominant thrombotic disorder. An increasing number of studies have shown that mutations in the SERPINC1 rs2227589 polymorphic site are correlated with a risk of venous thromboembolism (VTE) at common sites, such as lower extremity deep venous thrombosis and pulmonary thromboembolism. Currently, there are no reports of cerebral venous sinus thrombosis (CVST), a VTE site with a low incidence rate and rs2227589 polymorphism. CASE SUMMARY: Here, we report a Chinese CVST case with a mutation of the SERPINC1 rs2227589 polymorphic site, which did not cause significant AT deficiency. In a 50-year-old male patient presenting with multiple cerebral venous sinus thromboses no predisposing factors were detected, although a relative had a history of lower extremity deep venous thrombosis. We performed sequencing of the SERPINC1 gene for the patient and his daughter, which revealed the same heterozygous mutation at the rs2227589 polymorphic site: c.41+141G>A. CONCLUSION: The results showed that more studies should be conducted to assess the correlation between rs2227589 polymorphism and CVST.

Metformin therapy and prostate cancer risk: a meta-analysis of observational studies.

OBJECTIVE: Several observational studies have shown that metformin therapy may modify the risk of prostate cancer. We carried out a meta-analysis of relevant studies evaluating the effect of metformin therapy on prostate cancer risk. METHODS: We searched pubmed database (January 1966-February 2014) for case-control and cohort studies that assessed metformin therapy and prostate cancer risk. Two authors independently assessed eligibility and extracted data. Summary RRs was calculated using fixed-effects model or random-effects model. Heterogeneity among studies was examined using Q and I(2) statistics. RESULTS: We included six cohort studies and four case-control studies in the present meta-analysis, comprising 863,769 participants and 39,073 prostate cancer cases. The pooled RR of prostate cancer in relation to metformin therapy was 0.92 (95% CI: 0.84-1.02, P = 0.112). When we stratified the various studies by study type, we found that metformin therapy was associated with a significant reduced risk of prostate cancer among cohort studies (RR = 0.92, 95% CI [0.87, 0.96], P<0.001); however, no significant association was detected among case-control studies (RR = 0.95, 95% CI [0.78, 1.16], P = 0.632). There was also no indication of publication bias as suggested by Begg's test (P = 0.421) and Egger's test (P = 0.627). CONCLUSION: Our findings indicate that metformin therapy is not significantly associated with lower prostate cancer risk.

[Improving Agricultural Safety of Soils Contaminated with Polycyclic Aromatic Hydrocarbons by In Situ Bioremediation].

In order to reduce the risk of enrichment of polycyclic aromatic hydrocarbons (PAHs) in crops, reduce the potential hazards of food-sourced PAHs to human and increase the agricultural safety of PAHs contaminated soils, the bio-augmented removal of polycyclic aromatic hydrocarbons (PAHs) was investigated through in situ remediation by introducing Rhodobacter sphaeroides (RS) into the agricultural soil contaminated by PAHs. The 50-times diluted RS was sprayed on leaf surface (in area B) or irrigated to roots (in area D). The treatment of spraying water of the equal amount was taken as the control (A) and the wheat field without any treatment as the blank (CK). Treatments were conducted since wheat seeding. Soil and wheat samples were collected in the mature period to analyze the changes of community structure of the soil microorganisms and the concentration of PAHs in soils and investigate the strengthening and restoration effects of RS on PAHs contaminated soils. Compared to the CK Area, the areas B and D revealed that the variation ratio of phospholipid fatty acids (PLFAs) that were the biomarker of soil microorganisms was 29.6%, and the ratio of total PAHs removed was increased 1.59 times and 1.68 times, respectively. The dry weight of wheat grain of 50 spikes was increased by 8.95% and 12.5%, respectively, and the enrichment factor of total PAHs was decreased by 58.9% and 62.2% respectively in the wheat grains. All the results suggested that RS reduced enrichment of PAHs in wheat grains and increased wheat yield, which had great exploitation and utilization potentiality in repairing and improving the agricultural safety of the soils contaminated with PHAs.

Vasectomy and the risk of prostate cancer: a meta-analysis of cohort studies.

BACKGROUND: The relationship of vasectomy to prostate cancer has great public health significance. However, the results of observational studies were conflicting. To determine whether vasectomy is associated with the risk of prostate cancer, we performed a meta-analysis of cohort studies. METHODS: A literature search was carried out using Pubmed, Embase, Cochrane Libraryl, and China National Knowledge Infrastructure (CNKI) between January 1966 and July 2013. Before meta-analysis, between-study heterogeneity and publication bias were assessed using adequate statistical tests. Fixed-effect and random-effect models were used to estimate summary relative risks (RR) and the corresponding 95% confidence intervals (CIs). Potential sources of heterogeneity were detected by meta-regression. Subgroup analyses and sensitivity analysis were also performed. RESULTS: A total of nine cohort studies contributed to the analysis. There was heterogeneity among the studies but no publication bias. Pooled results indicated that vasectomy was not associated with a significant increase of total prostate cancer risk (RR = 1.07, 95% CI [0.79, 1.46]). When stratified the various studies by geographic location, we found a significant association between vasectomy and increased PCa risk among studies conducted in the USA (RR = 1.54, 95% CI [1.23, 1.93]), however, there was no significant association between vasectomy and PCa risk among studies conducted in non-USA countries (RR = 0.74, 95% CI [0.50, 1.09]). Furthermore, sensitivity analysis confirmed the stability of the results. CONCLUSIONS: In conclusion, the present meta-analysis of cohort studies suggested that vasectomy was not associated with increased risk of prostate cancer. More in-depth studies are warranted to report more detailed results, including stratified results by age at vasectomy, tumor grade, and tumor stage.

Association of metabolic syndrome and benign prostatic hyperplasia in Chinese patients of different age decades.

OBJECTIVES: To evaluate the relationship between metabolic syndrome (MetS) and annual prostate growth rates in Chinese patients of different age decades with benign prostatic hyperplasia (BPH). METHODS: We retrospectively analyzed the clinical data obtained from 1,052 Chinese men with BPH. Overnight fasting venous blood specimens were collected and serum levels of prostate-specific antigen, fasting blood glucose, high-density lipoprotein cholesterol, total cholesterol and triglyceride were recorded. We divided age into four groups: 50 ≤ age ≤ 60, 60 < age ≤ 70, 70 < age ≤ 80 and 80 < age ≤ 90. Pearson's correlation coefficient was used to test the linearity of the relationships between each of the MetS components and prostate volume and annual prostate growth rates generally and in different age decades. RESULTS: The median total prostate volume (69.01 ml) and median annual prostate growth rate (1.92 ml/year) were significantly higher in the MetS group compared with the non-MetS group (57.26 ml and 1.23 ml/year). Significant positive correlations were also found in total prostate volume and different age decades, while negative correlations were seen in annual prostate growth rate and different age decades. CONCLUSIONS: MetS is associated with an increased risk of total volume and annual prostate growth rate in BPH patients of different age decades.

[Comparison between transgenic insect-resistant cotton expressing Cry1Ac protein and its parental variety in rhizospheric fungal diversity].

The dynamics of rhizospheric fungal diversity and biomass at different sampling stages associated with two transgenic insectresistant cottons expressing Cry1Ac protein and their control varieties were studied under greenhouse conditions, followed by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real-time polymerase chain reaction (Q-PCR), in order to evaluate the ecological security of planting transgenic cotton expressing Cry1Ac protein. The results indicated that the fungal superior bands in rhizosphere of transgenic Bt cotton were similar with that of control cotton at four sampling stages, the more obvious difference in the blurred bands among transgenic Bt cotton, JM20 and SHIYUAN321 was detected. The rhizospheric fungal biomass of transgenic Bt cotton SGK321 was significantly lower than that of its parental control cotton at seedling stage, while the slight decrease in fungal biomass of transgenic Bt cotton XP188 was detected at boll forming stage, the ill-defined decrease, even growing tendency in two transgenic Bt cottons was detected at other stages. However, the difference of rhizospheric fungal community compositions and biomass was not only existed between transgenic cotton and its control, but also between SHIYUAN321 and JM20, and the same phenomenon was also detected between transgenic Bt cotton SGK321 and XP188. Hence, Bt protein is not the only incentive resulting in the difference in fungal community composition and diversity, the decrease in biomass between transgenic cotton and untransgenic cotton, different cotton varieties has an effect on them.

Suppression of bladder cancer growth by adeno-associated virus vector-mediated combination of HSV-TK and endostatin in vitro.

BACKGROUND: Gene therapy may offer a new tool for the treatment of bladder cancer. Previously, we have shown a significant antitumor effect in bladder cancer xenografts in a nude mouse model using intratumoral herpes simplex virus thymidine (HSV-TK) and endostatin gene monotherapy. OBJECTIVES: Given the high vascularity of human bladder cancer and the ability of HSV-TK or endostatin monotherapy to eradicate the tumors, we decided to test a novel combination of cytotoxic and antiangiogenic gene therapy using HSV-TK and endostatin adeno-associated viruses (AAV) in vitro. METHODS: We constructed the plasmid AAV-TK-IRES-Endostatin (pAAV-TIE) and packaged the AAV particles containing gene fragments of HSV-TK and endostatin. The combination anticancer effect of recombinant AAV-TIE (rAAV-TIE) was measured in vitro while rAAV-HSV-TK and rAAV-Endostatin were used as control groups. RESULTS: The inverted terminal repeat sequences were amplified using only one primer and the fragment between two ITRs of pAAV-TIE measuring about 4 kb, which indicated a stable sequence of pAAV-TIE. Three clear bands representing the AAV capsid proteins VP1, VP2, and VP3 could be seen on both lanes against a very low background, which demonstrated that chloroform extraction could effectively extract contaminants from rAAV stock without significant loss of the rAAV. In vitro, our results found that the transduction efficiency, measured from GFP-transduced tumors, was about 62%. The combination therapy led to an obvious apoptosis of bladder tumor cells compared with single HSV-TK or endostatin treatment. CONCLUSIONS: We concluded that the inhibition of angiogenesis using endostatin gene transfer, together with the cytotoxic HSV-TK gene therapy, resulted in a significant antitumor effect in vitro compared to the single gene based therapy in BTCC.

Potent antitumor activity of the combination of HSV-TK and endostatin by adeno-associated virus vector for bladder cancer in vivo.

BACKGROUND: Gene therapy may offer a new tool for the treatment of bladder cancer. Previously, we have shown a significant antitumor effect in bladder cancer xenografts in a nude mouse model using intratumoral herpes simplex virus thymidine (HSV-TK) and endostatin gene monotherapy. METHODS: Given the high vascularity of human bladder cancer and the ability of HSV-TK or endostatin monotherapy to eradicate the tumors, we decided to test a novel combination of cytotoxic and antiangiogenic gene therapy using intratumorally delivered HSV-TK and endostatin adeno-associated viruses (AAV). We constructed plasmid AAV-TK-IRES-Endostatin (pAAV-TIE) and packaged the AAV particles containing gene fragments of HSV-TK and endostatin. The combined anticancer effect of recombinant AAV-TIE (rAAV-TIE) was measured in vivo with rAAV-HSV-TK and rAAV-Endostatin as the control groups. RESULTS: The inverted terminal repeat sequence was amplified using only one primer and the fragment between two ITRs of pAAV-TIE measuring about 4 kb, which indicated a stable sequence of pAAV-TIE. Three clear bands representing the AAV capsid proteins VP1, VP2, and VP3 could be seen on both lanes against a very low background, which demonstrated that chloroform extraction could effectively extract contaminants from rAAV stock without significant loss of the rAAV. In vivo, our results showed that the tumors in mice injected with the rAAV-TIE not only took significantly longer to emerge but also that their growth, once established, was significant slower than that of tumors grown with single HSV-TK or endostatin treated animals. CONCLUSIONS: We concluded that the inhibition of angiogenesis using endostatin gene transfer, together with the cytotoxic HSV-TK gene therapy, resulted in a significant antitumor effect compared to the single gene based therapy in BTCC.

The adeno-associated virus-mediated HSV-TK/GCV suicide system: a potential strategy for the treatment of bladder carcinoma.

Novel treatment strategies such as gene therapy are warranted in view of the failure of current treatment approaches to cure a high percentage of patients with advanced bladder cancers. The emergence of cancer gene therapy potentially offers a number of exciting treatments. The majority of approaches involve strategies to suppress the function of activated oncogenes to restore the expression of functional tumour suppressor genes or to initiate tumour self-destruction. One gene therapy approach against tumours that holds great promise is suicide gene therapy. Herpes simplex virus thymidine kinase (HSV-TK) phosphorylates ganciclovir (GCV), which in turn interacts with cellular DNA polymerase and interferes with DNA synthesis to cause death of rapidly dividing cells. The development of an effective delivery system is absolutely critical to the usefulness and safety of gene therapy. At present, the adeno-associated virus (AAV) vector has the most promising potential in view of its non-pathogenicity, wide tropisms and long-term transgene expression in vivo. Gene therapy studies using different serotypes of recombinant AAV (rAAV) as delivery vehicles have proved rAAVs to be an effective modality of cancer gene therapy. In the present study, we investigated the suppression effect of AAV-mediated HSV-TK/GCV system on the bladder cancer cells and in mice xenograft models of bladder cancer. Our data demonstrate that rAAV-HSV-TK system controlled tumour cell growth and achieves strong antitumour efficacy in vivo. These findings provide a foundation for the development of potential targeted clinical therapies for bladder cancer in humans.

Potent antitumour activity of the combination of HSV-TK and endostatin armed oncolytic adeno-associated virus for bladder cancer in vitro and in vivo.

BACKGROUND: Gene therapy may offer a new tool for the treatment of bladder cancer. Previously, we have shown a significant antitumour effect in bladder cancer xenografts in a nude mouse model using intratumoural herpes simple virus thymidine (HSV-TK) and Endostatin gene therapy. OBJECTIVES: Given the high vascularity of human bladder cancer and the ability of HSV-TK or Endostatin monotherapy to eradicate the tumours, we decided to test a novel combination of cytotoxicity and antiangiogenisis gene therapy using intratumourally delivered HSV-TK and Endostatin adeno-associated viruses (AAVs). METHODS: We constructed plasmid AAV-TK-IRES-Endostatin (pAAV-TIE) and packaged the AAV particles contain gene fragments of HSV-TK and Endostatin. The function of HSV-TK and Endostatin was evaluated separately in vitro via T24 bladder tumour cells and human umbilical vein endothelial (HUVEC) cells. The combination anticancer effect of recombinant AAV-TIE (rAAV-TIE) was measured in vivo while rAAV-HSV-TK and rAAV-Endostatin as control groups. RESULTS: In vitro, rAAV-TIE was found to induce a significant increase in apoptosis in HUVEC cells equally as rAAV-Endostatin and confirmed that the inhibition of endothelial cells mediated by rAAV-TIE was associated with the apoptotic process. rAAV-TIE was found to induce a significant increase in apoptosis in T24 cells equally as rAAV-HSV-TK and confirmed that the inhibition of T24 cells mediated by rAAV-TIE was associated with the apoptotic process too. In vivo, our results showed that the tumours in mice injected with the rAAV-TIE not only took significantly longer to emerge but also that their growth, once established, was significant slower than that of tumours grown, compared with single HSV-TK or Endostatin treated animals. CONCLUSIONS: We concluded that the inhibition of angiogenesis using Endostatin gene transfer, together with the cytotoxicity HSV-TK gene therapy, resulted in a significant antitumour effect compared to the single gene based therapy in BTCC. The results warrant further development of the combination gene therapy, and suggest that this approach, directed towards systemic efficiency, could be used as an additional treatment for human BTCC.

Suppression of bladder cancer growth in mice by adeno-associated virus vector-mediated endostatin expression.

Novel treatment strategies such as gene therapy are warranted in view of the failure of current treatment approaches to cure a high percentage of patients with advanced bladder cancers. Testing of the hypothesis that blocking the angiogenic switch may keep tumour growth in check has been facilitated by the discovery of endogenous inhibitors of angiogenesis and has also added another research dimension to the field of cancer gene therapy. Consequently, the concept of targeting the tumour vasculature with anti-angiogenic agents has emerged as an attractive new strategy in the treatment of cancer. Targeted biological therapies that selectively interfere with tumour angiogenesis could improve survival among patients with bladder cancer. Endostatin is a tumour-derived angiogenesis inhibitor and is the first endogenous inhibitor of angiogenesis to be indentified in a matrix protein. Gene therapy represents an attractive approach to treat cancers and other chronic diseases. The development of an effective delivery system is absolutely critical to the usefulness and safety of gene therapy. At present, the adeno-associated virus (AAV) vector has the most promising potential in view of its non-pathogenicity, wide tropisms and long-term transgene expression in vivo. Gene therapy studies using different serotypes of recombinant AAV (rAAV) as delivery vehicles have proved rAAVs to be an effective modality of cancer gene therapy. In the present study, an IgG fragment was inserted at the start of the sequence coding for endostatin with the aim of enabling continuous secretion of endostatin the serum. We also investigated the suppression effect of AAV-mediated endostatin expression on endothelial cells and in mice xenograft models of bladder cancer. Our data demonstrates that rAAV-endostatin controlled tumour cell growth and achieves strong anti-tumour efficacy in vivo.

[Effects of double targeting gene therapy by suicide gene combined with endostatin gene in treatment of bladder cancer: an experimental study].

OBJECTIVE: To investigate the effect of double targeting gene therapy by using recombinant adeno-associated virus-thymidine kinase (TK)-internal ribosome entry site (IRES)-endostatin (ES) (rAAV-TIE). METHODS: Bladder cancer cells of the line T24 were cultured and transfected with rAAV-ES, rAAV-MCS (blank virus), and rAAV-TIE respectively. 72 hours later the levels of ES in the supernatants were measured by ELISA and annexin V apoptosis test kit was used to examine the apoptosis. Human umbilical vein endothelial cells (HUVECs) were transfected with rAAV-ES, rAAV-TIE, and rAAV-MCS respectively. MTT method and flow cytometry were used to detect the apoptosis of the HUVECs. Balb/c nude rats were inoculated subcutaneously with T24 cells. Twenty rats with tumor were randomly divided into 4 equal groups to be treated by rAAV-MCS, rAAV-TK, rAAV-ES, or rAAV-TIE, and 5 rats were used as control group. Four weeks later, blood samples were collected to detect the ES level by ELISA. The tumors were taken out to undergo microscopy to calculate the microvessel density(MVD). RESULTS: 72 h after transfection, ES could be detected in the supernatants of the T24 cells transfected with rAAV-ES, and rAAV-TIE. The apoptotic rates of the T24 cells transfected with rAAV-TK and rAAV-TIE were 34.12% and 36.91% respectively, significantly higher than those of the T24 cells transfected with rAAV-MCS and of the control group (3.08% and 0.84%, all P < 0.05). Transfection of rAAV-ES and rAAV-TIE increased the apoptotic rate of the HUVECs time-dependently. Nine days after the transfection rAAV-ES, rAAV-TK, rAAV-TIE, the tumor volumes of the rAAV-ES, rAAV-TK, and rAAV-TIE groups were (0.75 +/- 0.08), (0.71 +/- 0.11), and (0.52 +/- 0.09) cm(3) respectively, all significantly lower than those of the rAAV-MCS group and control group [(1.27 +/- 0.13) and (1.24 +/- 0.17) cm(3) respectively, all P < 0.05]. CONCLUSION: rAAV-TIE effectively inhibits the tumorigenesis and angiogenesis in bladder cancer. Double targeting gene therapy against bladder cancer can be achieved by using rAAV.

[Correlation of Ureaplasma urealyticum and Chlamydia trachomatis infections with male sterility: a meta-analysis of randomized control trials].

OBJECTIVE: To evaluate the correlation of Ureaplasma urealyticum (Uu) and Chlamydia trachomatis (Ct) infections with male sterility. METHODS: Data from CBMA and CNKI were searched and studies were made for the correlation of Uu and Ct infections with male sterility by retrieval strategy worked out according to the Collaborative Review Group search strategy. The results were expressed as odds ratio (OR) and 95% confidence intervals (95% CI). The results of 28 clinical controlled trials from 1994 to 2005 were analyzed by software RevMan 4.2. Odds ratio (OR) was applied to the evaluation of the correlation between Uu infection and male sterility. RESULTS: Eighty-eight relative trials were retrieved, of which 28 were included in the Meta-analysis. The combined ORs of Uu and Ct infections to male sterility were OR(Uu): 4.73 (95% CI: 3.77-5.94) and OR(Ct): 4.59 (95% CI: 3.24-6.50). CONCLUSION: Uu and Ct infections are very important risk factors of sterility in Chinese men.

Can intravesical bacillus Calmette-Guérin reduce recurrence in patients with superficial bladder cancer? A meta-analysis of randomized trials.

OBJECTIVES: To determine whether intravesical bacillus Calmette-Guérin (BCG) administration reduces recurrence after transurethral resection of superficial bladder cancer using a meta-analysis. METHODS: Published data of randomized clinical trials comparing transurethral resection plus intravesical BCG to either resection alone or resection plus another treatment were analyzed, considering possible confounding factors such as disease type, maintenance therapy, and others. Both the fixed effect model and the randomized effect model were applied, and the odds ratio (OR) with its 95% confidence interval (CI) was used as the effect size estimate. RESULTS: We searched 176 trials, eliminated 151 of them, and identified 25 trials with recurrence information on 4767 patients. Of 2342 patients undergoing BCG therapy, 949 (40.5%) had tumor recurrence compared with 1205 (49.7%) of 2425 patients in the non-BCG group. In the combined results, a statistically significant difference in the OR for tumor recurrence between the BCG and no BCG-treated groups was found (randomized combined effect OR 0.61, 95% CI 0.46 to 0.80, P <0.0001). Stratified by BCG maintenance and disease type, the combined results of the individual reports showed statistical significance for BCG maintenance (OR 0.47, 95% CI 0.28 to 0.78, P = 0.004) and treatment of papillary carcinoma (OR 0.50, 95% CI 0.33 to 0.75, P = 0.0008). Chemotherapy and BCG plus chemotherapy/immunotherapy were not better than BCG alone. CONCLUSIONS: Adjuvant intravesical BCG with maintenance treatment is effective for the prophylaxis of tumor recurrence in superficial bladder cancer. For patients with papillary carcinoma, adjuvant intravesical BCG with maintenance therapy should be offered as the treatment of choice.