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In this study, voltage was used as a disturbance factor to investigate the relationship between microbial community and methane (CH4) production flux in a microbial electrolytic cell coupled anaerobic digestion (MEC-AD). Metabolic flux analysis (MFA) was used to explore the relationship between the CH4 metabolic flux produced and the microbes. The results showed that both methane production flux and hydrogen production flux changed significantly upon voltage disturbance, while the voltage disturbance had little effect on acetic acid production flux. The maximum CH4 production flux under 0.6 V disturbance was 0.522±0.051, which increased by 77% and 32%, respectively, compared with that of the control group under 1.0 V (0.295±0.013) and under 1.4 V (0.395±0.029). In addition, an average of 15.7%±2.9% of H2 (flux) was used to reduce CO2 to produce CH4 and acetic acid, and an average of 27.7%±6.9% of acetic acid (flux) was converted to CH4. Moreover, the abundance of Lachnospiraceae significantly affected the flux of acetic acid. The flux of CH4 production is positively correlated with the abundances of Petrimonas, Syntrophomonas, Blvii28, and Acinetobacter, and negatively correlated with the abundances of Tuzzerella and Sphaerochaeta. The species that affected the flux of H2 and CH4 were similar, mostly belonging to Bacteroides, Clostridium, Pseudomonas and Firmicutes. Furthermore, the interspecies interaction is also an important factor affecting the MEC-AD methanogenesis flux.
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Acetates , Anaerobiosis , Bioreactors , Electrolysis , MethaneABSTRACT
Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase-9, Bax mRNA and protein were tested by RT-PCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase-9 and Bax mRNA and protein were significantly higher than those of other two groups (P<0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the down-regulation of BCR-ABL and the up-regulation of caspase-9 and Bax.
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Objective:To evaluate the safety and clinical effect of holmium laser enucleation of the prostate (HoLEP) for pati-ents with benign prostatic hyperplasia (BPH) .Methods:A total of 40 patients with BPH from Jan 2015 to Jul 2015 were underwent HoLEP.Pre-and postoperative complication were recorded and the differences of the clinical parameters were also compared, including peak urinary flow rate (Qmax), residual urine volume (PVR), International Prostate Symptom Score (IPSS), quality-of-life (QOL) .Results:A total of 34 patients were included after operation and 6 patients lost to follow-up. Qmax, PVR, IPSS, QOL after three months postoperatively were improved significantly compared with that of preoperative (P<0.01) .Complications included intraoperative bladder injury (1 case), postoperative stress incontinence (2 cases), urethral stricture (1 case) .No patients experienced the sign of TUR syndrome and blood transfusion for severe bleeding.Conclusion:HoLEP has the advantage of good therapeutic effects,wide scope of application,better safety performance,smaller risk and it’s worth popularizing in primary hospitals.
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Objective To explore the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and drug resistance of leukemic cell line NB4.Methods NB4 cells were divided into three groups: interference group, negative control group and untreated group.The infection efficiency of lentivirus for NB4 cells was detected by flow cytometry, and the expression of HOXA10 gene of NB4 cells at mRNA and protein level was detected by real-time PCR and Western blot.Cell survival was determined by MTF assay, and apoptosis and necrosis rates were detected by flow cytometry.Western blot was used to detect the influence of down-regulation HOXA10 gene on the multi-drug resistance-1 (MDR-1) protein.Results The ratio of GFP positive cells was up to 90 ％.HOXA10 gene mRNA and protein levels were decreased in interference group compared with control group.The inhibition rate of interference group was (52.12±4.02) ％, the apoptosis rate of interference group was (30.0±2.7) ％, and their differences in the interference group and in control groups (negative control group and untreated group) were significant (P ＜ 0.05).Western blot results showed that interfering HOXA10 gene significantly reduced the resistance gene MDR-1 expression level and reverse the drug-resistant of leukemia cells.Conclusions Lentivirns-shHOXA10 can steadily reduce the expression level of HOXA10, inhibit the leukemic cells proliferation, promote apoptosis and reverse drug-resistant.HOXA10 gene is expected to become a new target for reversing leukemia drug resistance.
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Objective To explore the prognostic value of N-terminal pro-brain natriuretic peptide ( NT-pro-BNP) levels in critically ill infants. Methods Eighty-one critically ill infants were enrolled from January 2013 to January 2014 in pediatric intensive care unit. The minimum of pediatric critical illness score ( PCIS) and the number of dysfunction organs were calculated within 24 hour after admission. According to PCIS,the critically ill infants were divided into extremely critical group(PCIS≤70,n=25),critical group (PCIS 71-80,n=30)and non-critical group(PCIS>80,n=26). According to the prognosis,the critically ill infants were divided into survival group (n=68)and death group(n=13). The serum NT-pro-BNP levels were determined on the first day,third day and convalescent phase. The relationships of serum NT-pro-BNP levels with PCIS and the number of dysfunction organs and prognosis were observed. Results The study showed statistical significances of serum NT-pro-BNP levels among the extremely critical group, critical group and non-critical group,whether on the first day,or on the third day and convalescent phase(P<0. 01). There were statistical significances of serum NT-pro-BNP levels among different stages of the disease in each group(P<0. 01). Compared with survival group,PCIS was significantly lower and the serum NT-pro-BNP levels and the number of dysfunction organs were significantly higher in death group. The serum NT-pro-BNP level on the third day was higher than that on the first day in death group ( P<0. 01 ) , while no significant difference was found in survival group. The serum NT-pro-BNP levels on the first day and the third day and PCIS were negatively correlated(r= -0. 59,P<0. 01;r= -0. 66,P<0. 01). The serum NT-pro-BNP levels on the first day and the third day and the number of dysfunction organs were positively correlated(r=0. 40,P<0. 05;r=0. 57,P<0. 01). Conclusion The serum NT-pro-BNP levels of the critically ill infants are correlated with disease severity,and can be useful for assessing the severity of critical illness.
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ObjectiveTo investigate the diagnostic value of N-terminal pro-brain natriuretic peptide (NT-proBNP) and MB isoenzyme of creatine kinase (CK-MB) for heart failure (HF) in pneumonia children.MethodsThe NT-proBNP and CK-MB were assayed in 132 pneumonia children with HF, 138 pneumonia children without HF and 62 healthy children were recruited into this study. A receiver operating characteristics (ROC) curve and a logistic regression model were employed to assess the diagnostic accuracy of NT-proBNP and CK-MB for HF in pneumonia children.ResultsPneumonia children with HF had higher blood NT-proBNP and CK-MB than those in pneumonia children without HF and healthy controls (P<0.01 for both). Pneumonia children with HF had higher blood NT-proBNP and CK-MB than the pneumonia children without HF. The area under curves (AUCs) of NT-proBNP and CK-MB for HF were 0.85 and 0.72, respectively. The AUC for their combinational usage was 0.87.ConclusionBoth NT-proBNP and CK-MB are effective markers as diagnostic adjuncts for HF in pneumonia children. Combination of NT-proBNP and CK-MB can improve the diagnostic accuracy for HF in pneumonia children.
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Objective To investigate the effect of rivaroxaban on prevention from deep venous thrombosis after hip replacement surgery .Methods 80 patients with lower limb parallel arthroplasty were randomly divided into observation group and control group from January 2014 to January 2015.40 cases in each group.Control group was given aspirin treatment, observation group was given rivaroxaban treatment, indicators changes were followed up and recorded.Results PT, APTT values of observation group one week after treatment were (35.79 ±2.16),(15.84 ±2.11) s, which were better than control group (45.64 ±2.12),(18.72 ±2.02) s (P<0.05).TT, D-dimer of observation group(13.48 ±2.10)s,(2.34 ±0.33)g/L after a week were better,than control group (16.38 ±1.79)s, (1.42 ±0.21)g/L (P<0.05) respectively.Conclusion Effect of rivaroxaban in prevent deep venous thrombosis after hip replacement surgery is exact without obvious adverses reaction, it is worthy of further research and application.
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Objective To screen siRNAs that can effectively inhibit Apollon gene expression and determine the cellular functions of those siRNAs.Methods A chemical synthesis method was used to synthesize 3 siRNA sequences against different sites of Apollon.They were transfected into the human breast cancer MCF-7 cells by using Lipofectamine 2000.mRNA level of Apollon was determined by reverse transcription-polymerase chain reaction (RT-PCR).Cellular immunity fluorescence quantitative analysis combined with confocal laser technology was used to determine the protein level of Apollon.Methyl thiazolyl tetrazolium bromide (MTT) assay and flow cytometry were used to determine the effects of siRNA targeting Apollon on proliferation and apoptosis of MCF-7 cells,respectively.Results Three pairs of siRNA could significantly inhibit Apollon mRNA expression,at the inhibition rates of (36.201±11.629) ％,(67.308±7.686) ％and (47.123±12.000) ％,respectively (P ＜ 0.05).After tranfection by siRNA2,Apollon protein fluorescence intensity was (14.97±2.08) ％ compared with control cells.The cell proliferation MCF-7 was inhibited by (73.361±2.118) ％and apoptosis was increased by (28.793±0.743) ％.Conclusions Screened siRNA2 effectively silences Apollon gene expression,effectively inhibits the proliferation and increases the apoptosis of MCF-7 cells.This provids the foundation for its clinical application in cancer therapy.
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Objective To design and screen small interefere RNA (siRNA) targeting of HOXA7,and to investigate the effect of the siRNA on human lung cancer LETP-a-2 cells proliferation and apoptosis in vitro.Methods Three pairs of siRNA targeting of HOXA7 and one pair of siRNA for negative control were transfected respectively into LETP-a-2 cells through cationic liposome.The mRNA and proteion expression levels of HOXA7 were observed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The effect of HOXA7 siRNA on growth and apoptosis of LETP-a-2 cells were measured by MTT and flow cytometry.Results All the three pairs of siRNA could inhibit HOXA7 expression effectively,among which siRNA2 got the best effects,the silence rates were (57.344±4.743) ％ on mRNA level and (52.219±0.550) ％ on protein leval.The proliferation was inhibited and the apoptosis was promoted by the siRNA targeting HOXA7 in LETP-a-2 cells,among which siRNA2 got the favourite results,the inhibitory rate was (48.144±4.992) ％ and the apoptosis rate was (26.613±0.612) ％.Conclusion The siRNA2 targeting of HOXA7 enrolls in promoting apoptosis and inhibiting grows of LETP-a-2 cells,indicating that manipulation of HOXA7 expression may be a potential therapeutic strategy for cancer.
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Background and purpose:Apollon gene is highly expressed in leukemia and other tumors. The study aimed to discuss whether RNAi technology can reverse multidrug resistance of chronic myeloid leukemia cell line K562 through constructing a eukaryotic vector of short hairpin RNA (shRNA) targeting at Apollon gene. Methods:The eukaryotic vector pGPHI-GFP-Neo-Apollon with shRNA targeting at Apollon gene was constructed and then transfected into K562 cells by LipofectamineTM2000, and G418 pressure selection. Reverse transcription-polymerase chain reaction (RT-PCR) and immunolfuorescence were used to detect the expression of Apollon mRNA and protein after Apollon was transfected stably in K562 cells. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP16) after transfection with shRNA-Apollon were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: pGPHI-GFP-Neo-Apollon carrier was constructed successfully and expressed stably in K562 cells, and after G418 screening, it silenced Apollon mRNA and protein expression effectively. According to the result of MTT, the sensitivity of K562 cells to VCR and VP16 increased significantly in the group of gene interference, with half of its inhibition concentration (half-inhibitory, IC50) value signiifcantly lower than the control group (P0.05). Conclusion:pGPHI-GFP-Neo-Apollon carrier can enhance the abilities of VCR and VP16 to induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells, it is hinted that RNA interference targeting Apollon gene may reverse the multidrug resistance of leukemia cells in some degree.
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Objective To investigate the effects of small interference RNA (siRNA) targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cell line.Methods Effective and specific siRNA oligo targeting HOXA9 was designed and compounded.It was transfected transiently into U937 cells by cationic liposome.The cells was divided into three groups:experimental group(siRNA targeting HOXA9 was transfected by liposome),negative control group (negative siRNA was transfected by liposome) and cell control group (add equal cells and medium).The expression of HOXA9 mRNA and protein were detected by reverse transcription PCR and Western blot.The cell proliferation was assessed by MTT.The apoptosis of each group were measured by Annexin V-FITC.Results Aftcr transfected by siRNA targeting HOXA9,the relative mRNA expression levels of HOXA9 in the experimental group,negative control group and cell control group were (22.980±0.548) ％,(82.371±1.517) ％ and (84.637±2.252) ％,respectively (P ＜ 0.05),and the relative protein expression levels were (50.377±2.773).％,(105.500±3.900) ％ and (111.392±3.905) ％,respectively (P ＜ 0.05).The inhibitory rates of cell proliferation and the apoptosis rates of the experimental group were significantly increased.The inhibitory rates of cell proliferation of 24 h,48 h and 72 h were (41.909±4.333) ％,(54.470±3.756) ％ and (65.835±1.024) ％,respectively,and the apoatosis rate was (26.800±2.081) ％.Compared with 2 controls,the experimental group differences had statistically significance (P ＜ 0.05).Conclusion siRNA targeting HOXA9 can effectively silence HOXA9 gene expression in U937 cell,suppress cell proliferation and induce cell apoptosis obviously,which providing experimental basis for clinical lenkemia therapy by targeting HOXA9 gene.
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Objective To obtain effective siRNA fragment of HOXA10 gene and verify its function,to supply experimental evidence for tumor prevention and curation by RNAi targeting to HOXA10 gene.Methods Three pairs of small interfering RNA targeting to the different sites of HOXA10 were designed and introduced into A549.The mRNA expression of HOXA10 of A549 was detected by semi-quantitative RT-PCR,the cell proliferation was assayed by MTT,the apoptosis was measured by flow cytometry.The most effective siRNA assay was screened and was tested the relationship between it and proliferation and apoptosis.Results The mRNA of HOXA10 was inhibited by three siRNAs in A549 cells,among which siRNA1 gave the strongest inhibiting of HOXA10 by ODR was (20.190±1.698) ％.The inhibitory rate of cell proliferation was (69.793±2.092) ％ and the apoptosis rate was (29.593±2.670) ％.Conclusion siRNA1 can specifically degrade HOXA10 mRNA and inhibit the proliferation of A549 cell and promote its apoptosis.
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Objective To investigate the effects of Livin antisense oligonucleotide (ASODN) on the proliferation and apoptosis of human leukemia (K562) cells. Methods Specific phosphorothioate ASODN and missense oligonucleotide (MSODN) target Livin mRNA were synthesized and transfected into K562 cells following cationic liposome. The proliferation inhibition of K562 cells was assessed by MTT. The apoptosis rate of each group was detected by Annexin V-FITC. The expression of Livin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Results ASODN at a final concentration of 600 nmol/Lcould inhibit the K562 cells proliferation (IR) was (52.99t2.67) % and the expressions of Livin mRNA (ODR)was (59.75±3.24) %, the apoptosis rate was apparently increased [(36.89±1.08) %] (P ＜0.01); but the difference between Lip-MSODN group, Lip control group and cell control group was not statistically significant (P ＞0.05).Conclusion Livin ASODN may decrease Livin gene expression, suppress K562 cells proliferation effectively, and induce significant apoptosis of K562 cells.
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Objective To investigate the influence of very late antigen-4 (VLA-4) antibody and VLA-4 antibody combined with VP_(16) in vitro on childhood leukemic cells' apoptosis and explore the protection of bone marrow stromal cells (BMSC) upon leukemic cells and its related mechanisms. Methods Leukemic bone marrow stromal cells were isolated by human lymphocyte separation medium and in vitro culture of BMSC (adherent) and leukemia cells (suspended) BMSC+leukemic cells group were as control. Then VLA-4 antibody and/or VP_(16) were added respectively to VLA-4 antibody group, VP_(16) group and VLA-4 antibody combined with VP_(16) group to detect the apoptosis of leukemic cells in different groups through Annexin V-FITC double-labeled flow cytometry and the expression of Survivin, bcl-2 genes in each group of leukemic cells detected by RT-PCR. Results The results showed by flow cytometry that compared with the control groups, for 12 h or 24 h, the early and total apoptosis rates of leukemic cells of the three experimental groups were significantly increased(P <0.05); the early and total apoptosis rates of leukemic cells treated with VLA-4 antibody combined with VP_(16) group was markedly increased, compared with the control group (P <0.05); the comparison of the early and total apoptosis rates for the three experimental groups between 12 h and 24 h was significantly different (P<0.01). Moreover, RT-PCR results showed that the expression of Survivin and bcl-2 genes of leukemic cells in three experimental groups was reduced in varying degrees and the reduction of VLA-4 antibody combined with VP_(16) group was the most obvious. Conclusion BMSC plays a protective role on leukemic cells, and VLA-4 antibody can block the adhesion between BMSC and leukemic cells promoting leukemic cells apoptosis and enhance the sensibility of apoptosis of leukemic cells induced by chemotherapeutics.
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Objective To study the effect of fish oil on calcium oxalate kidney stone formation. Methods The efficacy of unprocessed fish oil in the treatment of 20 hypercakiuric patients with urinary stones was evaluated, after administration of fish oil at dosage of 3000mg daioy of one week. Results Excretion of urinary calcium reduced significantly from (461.37?80.67)mg/d to (312.63?40.57)mg/d.The Mg2+ reduced from (136. 41?56. 12) mg/d to (108.43?18.96) mg/d. The Mg2+ /Ca2+ increased significantly from (0.29?0.08) to ( 0. 36?0.11)-Uric acid remained unchanged. Conclusion Unprocessed fish oil is safe and effective for the reduction of urinary calcium and might be used for the prevention of calcium stone recurrence.
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Cataract surgery is now often referred to as refractive surgery.It has come into the focus in recent years to choose the intraocular lens of appropriate power so as to achieve the expected postoperative refraction.Two main factors influence the measurement of intraocular lens power(IOL),the IOL power calculation formula and biological measurement of the ocular.The achievement of expected surgical outcome depends on the selection of the IOL power calculation formula and biological measurement instrument that suit the intraocular lens the specific patient.The article presents an overview of IOL power calculation with a focus on the above two aspects.
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Objective To investigate therapeutic effects of transurethral ureteroscopic pneumatic lithotripsy(TUPL).Methods A retrospective analysis was made on 320 cases of ureteral calculus treated with TUPL from September 2002 to August 2005.A Wolf F9.8 rigid ureteroscope and an EMS pneumatic ballistic lithotriptor were used.The probe was 0.8 mm or 1.0 mm in diameter.The pulse settings was 200~300 kPa.The fragmentation was conducted with single or multiple episode of treatment.Results The stone was pushed back to the renal pelvis in 6 cases and a conversion to extracorporeal shock wave lithotripsy(ESWL) was needed.The insertion of the ureteroscope was failed in 5 cases and a conversion to open surgery was required.The TUPL time was 2~10 min(mean,5.5 min) and the intraoperative blood loss was 3~15 ml(mean,7 ml).The stones were successfully fragmented on one session in 309 cases,the success rate being 96.6%(309/320).The length of postoperative hospital stay was 3~7 d(mean,4.5 d).There were 24 cases of mucosal injury of the ureter and 4 cases of false passage,all of which were cured with double-J catheter indwelling for 1 month.Follow-up examinations for 1~10 months(mean,4 months) showed stone clearance within 1 month and no ureteral stenosis or urinary infection.Conclusions Transurethral ureteroscopic pneumatic lithotripsy for the treatment of ureteral calculus is safe and effective.
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Objective To study the effect and safety of radiofrequency ablation (RFA) in the treatment of liver cancer.Methods The clinical data of prospectively and non-randomly selected 102 liver cancer patients were analysed. Results 102 patients with a total of 216 tumors were found in this series, the size of hepatomas were 1.5 to 14*!cm in diameter (average 4.9*!cm in diameter). In 67 of primary liver hepatocellular carcinoma, the tumors' size were smaller than 5*!cm in diameter in 21 cases; 35 with secondary liver tumors. RFA were performed under the guidance of ultrasonography in 43 cases, under CT in 50, under laparoscopy in 3, and 6 were performed during open surgery. All patients received ultrasonography and CT scanning of liver one month after the ablaton. The echoes of the tumors were stronger, and the area of the echo reached was larger than pre-ablation; blood flow in the tumor obviously reduced and even disappeared in sonography; CT showed that the shadow density in the tumor was diminished. Of the 102 patients, 14 underwent 18-FDG-PET imaging 30 days after the ablation, of them, 11 showed defect of radiation in the tumor, 3 showed residual locus in the tumor. 2 of the 3 cases underwent RFA again and the residual was disappeared completely, the another subject to ethanol injection because of advanced age. All the patients were followed up for 3-18 months. Except 6 patients with late metastastic liver cancer died within 1 year after RFA, the other 96 cases remained alive now, the 1-year survival rate was 94.1%. Conclusions RFA is a new ideal therapy for liver cancer. It is safe, effective, and good tolerance with little trauma.
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Objective To investigate the therapeutic effect of herceptin to rat bladder cancer. Methods Totally 50 SD rats were randomly divided into experimental group (n=40) and control group (n=10). The rats of experimental group received a perfusion into the bladder with N-methyl-N-nitrosourea (MNU) at 2.5 mg every time, once per 3 weeks for 4 times in all, while those of the control group underwent a perfusion with normal saline instead of MNU. In 1 week after the last perfusion, the bladders were slited for gross observation and the biopsies were carried out for pathological observation with HE staining. Immunohistochemical SP method and RT-PCR were applied to detect HER-2 protein and mRNA expression in tumor tissues. Then 16 rats with HER-2 positive bladder cancer were randomly and equally divided into treatment group (herceptin, 20 mg/kg by intraperitoneal injection, once per week for 6 weeks) and non-treatment group (normal saline, 0.5 ml). Tumor weight, size and shape were measured and calculated for the inhibitory rate of herceptin. HE staining was carried out for the morphology of the tumor mass. The expression of HER-2 protein was detected with immunohistochemical SP method, and apoptosis in bladder tumor tissues with TUNEL method. Results The results of inhibitory rate was 50.0%, and it showed that the differences were statistically significant in treatment group compared with non-treatment group (P