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In recent years, adaptive laboratory evolution (ALE) has emerged as a powerful tool for basic research in microbiology (e.g., molecular mechanisms of microbial evolution) and efforts on evolutionary engineering of microbial strains (e.g., accelerated evolution of industrial strains by bringing beneficial mutations). The ongoing rapid development of next-generation sequencing platforms has provided novel insights into growth kinetics and metabolism of microbes, and thus led to great advances of this technique. In this review, we summarize recent advances in the applications of long-term and short-term ALE techniques mainly for microbial strain engineering, and different modes of ALE are also introduced. Furthermore, we discuss the current limitations of ALE and potential solutions. We believe that the information reviewed here will make a significant contribution to further advancement of ALE.
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High-Throughput Nucleotide Sequencing , Laboratories , MutationABSTRACT
Objective To explore the expression and clinical value of major histocompatibility complex class-Ⅰ related chain A (sMICA) molecule in serum of patients with renal tumor.Methods From March 2013 to July 2013,60 patients with renal tumor,including 37 male patients and 23 female patients were enrolled in this study as experimental group.The mean age was 46 years (range 34-76 years).The pathological diagnosis included renal cell carcinoma in 48 cases and renal angiomyolipoma in 12 cases.The stage classification included T1 stage in 20 cases,T2 stage in 14 cases,T3 stage in 10 cases and T4 stage in 4 cases.Lymphatic metastases were found in 11 cases and metastases in other organs were found in 4 cases.Another 20 healthy volunteers were enrolled as control group,including 10 male and 10 female.The mean age was 31 years (range 24-50 years).The ELISA method was used to detect the soluble MICA's (sMICA) level in serum.And the results were compared with tumor's malice,TNM pathology stages,metastasis.In 15cases with renal cell carcinoma,the expression of MICA molecule in tumor masses and paraneoplastic masses was measured by immunohistochemical (IHC) method.The quantitative expression of MICA-mRNA was detected by RT-PCR in 9 tumor masses and 3 paraneoplastic masses.Results The level of sMICA in renal malignant tumor group was (348.5±32.5) pg/ml,while the sMICA's level in benign renal tumor groups was (289.3±30.4) pg/ml and that in the control group was (168.4±43.2) pg/ml.The level of sMICA in malignant group is statistically higher than that in benign group and control group (P＜0.05).The level of sMICA in T1 、T2 、T3 and T4 stage was (304.3±27.4),(308.4±26.8),(368.3±33.4),(378.4±43.4) pg/ml,respectively.Insignificant difference only demonstrated between T1 and T2 stage.The level of sMICA in those patients with and without lymphatic metastasis was (326.2±32.4),(319.4±32.5) pg/ml,respectively (P＞0.05).Significant difference in the sMICA level could also be observed between patients with other organ metastasis (373.4±45.4) pg/ml and those without metastasis (346.4±31.5) pg/ml (P＜0.05).The IHC results revealed that high expression of MICA molecule in tumor cell.However,this oppsite result was demonstrated in cells located in paraneoplastic tissues.In the results of RT-PCR,the MICA-mRNA level (2.03) in tumor masses was significantly higher than that in pareneoplastic masses (0.77) (P＜0.05).Conclusions MICA highly expressed in renal tumor,and its expression correlates with tumor's malice,TNM pathologic stages,and metastasis.
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Objective To explore renal transplantation model in non-human primate cynomolgus monkeys.Methods 50 non-human primates' kidneys were transplanted into the lower part of the abdomen with end-to-side anastomosis of renal artery to aorta and renal vein to inferior vena eava,and with end-to-end anastomosis of ureter to bladder.Results In the 50 cases,1 case death as accident of anesthesia;7 cases with postoperative complications,and all with creatinine sudden rise,after ultrasonic examinations showed that 2 cases with renal vein thrombosis,and 5 cases appeared urinary leakage.All animal models were without surgical infections,and with normal serum creatinine,urine output.Conclusion Non-human primate animal kidney transplantation model establishment method is reliable,but should pay attention to the the surgical technique training,complications prevention.The model is valuable for application in the research of immune tolerance,heterogeneous transplant.
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ObjectiveTo observe the effect of rapamycin on the proliferation and cell cycle of prostate cancer cell lines PC3,and to investigate the mechanism that it inhibits the growth and metastasis of prostate cancer cells.MethodsPC3 cells were cultured in vitro,and treated with different concentrations (10,25ng/ml) of cycle raparnycine.MTT was used to measure the change of proliferation of PC3 cells.Flow cytometry was used to measure the changes of cell cycle and apoptosis of PC3 cells.Nude mice were used to detect the effect of rapamycin on metastasis.ResultsThe proliferation of PC3 cells was significantly inhibited by rapamycin,and a time- and concentration-dependent relationship was shown,the inhibited rate was(42.23 ± 0.78 ) ％ after 36 h in the group of 25 ng/ml ( P ＜ 0.05 ).Flow cytometry analysis showed rapamycin significantly inhibited the cell cycle,prompted the apoptosis,and increased the number of cells in G0/G1 phase at 36 h with a rate of cell staying at Go/G1 [ (92.17 ± 0.69 ) ％ ] ( P ＜ 0.05 ).The weight of tumors in nude mice in the control group was significantly greater than that in RPM group[ (3.41± 0.28)g vs ( 1.19 ± 0.23 )g] ( P＜ 0.05 ),and metastatic sites of the lung and liver in the control group were significantly mote than the RPM group [ 100％ (7/7) vs 14.29％ ( 1/7 ) ] ( P ＜ 0.05 ).Conclusions Rapamycin significantly inhibits the proliferation and metastasis of osteosarcoma.The rapamycin-based regimen is valuable for clinical application.
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Objective To evaluate the safety and efficacy of colour doppler in guiding the percutaneous nephrolithotomy(PCNL) for Staghom calculi.Methods The clinical records of 46 patients with renal calculi who underwent PCNL were retrospectively analyzed.Patients' mean age was 43 years old,and the range of diameter of stone was 3.0 -7.5 cm.Among these cases,29 cases had complete staghorn calculus.One case had isolated kidney stone.And the other 6 patients had open surgery history.Using Colour Doppler guidance,the percutaneous nephrolithotomy for renal calculi was conducted.Results F22 percutaneous channel was successfully established in 55 sides of 46 patients,with the first and the second phase surgeries of 41 and 14 sides respectively.Single-,double- and three-channel PCNL were performed in 38,16,and 1 sides respectively.Abdominal x-ray conducted at 3 -4 days post operation revealed residual stones on 18 sides,with the range of 0.4 -2.0 cm in diameter.Second-phase lithotripsy was conducted on 14 sides of patients.After the first and the second phases of surgery,4 sides having residual stones ranging 0.6 - 1.0 cm,underwent extracorporeal shock wave lithotripsy.Ater the third-phase surgery,the diameters of residual stones were much less than 0.5 cm and patients were treated with medication.The total rate of clearance was 87.0％ (48/55).The duration of surgery was 65 - 160 minutes and 95 minutes on average.One patient having delayed bleeding was cured with selective renal artery embolization.There were no complications such as nephrectomy,deaths,pleural or intestinal damage during the period of study.Patients were followed up for 3 to 18 months.Six of 9 patients with renal insufficiency recovered after surgery.The serum creatinine in the remaining 3 patients ranged 185 -220 μmol/L Conclusion Colour Doppler-guided percutaneous nephrolithotomy for renal calculi is safe and effective.
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Objective To investigate the treatment of recipients carrying hepatic virus after renal transplantation. Methods Of the 14 patients, 8 carried HBV and 4 HCV, and the other 2 both HBV and HCV. HBV DNA or RNA was negative before transplantation.Results During the follow-up period of 3 to 20 months, 10 patients had liver dysfunction with higher ALT and AST, but negative for HBV DNA and/or HCV RNA. After adjusting the dosage of immunosuppressants and treatment of liver protection, liver function of these patients all restored to normal level.Conclusion In the hepatic virus carriers receiving renal transplantation, liver dysfunction caused by drug-induced liver damage or hepatic viral injury should be distinguished and corresponding treatment should be given in time.
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Objective To investigate the expression of complement split product C4d and its significance in renal allograf tissue of chronic rejection in rats.Methods The healthy closed population Wister rats and SD rats were used as donator and acceptor in renal transplantation.The chronic rejection model of renal transplant in rats was established and the rats were divided into 2 groups.The rats in experimental group were given Mycophenolate mofetil(MMF)(10 mg/kg) and those of the control group were given nothing except CsA(5 mg?kg~(-1)?d~(-1)) for 10 days.At the 12th week of renal transplantation,the allograft was tested by light microscope,and the pathological changes of renal grafts and the expression of C4d in peritubular capillaries were observed.Results On the 12th week of renal transplantation,the morphology changes of chronic rejection was observed in the experimental group and obvious C4d deposition was detected in peritubular capillaries of renal allograft tissue,with significant difference compared with those of the control group(all P
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Objective To explore the significance of complement split product C4d in monitoring chronic rejection of renal allografts in rats.Methods Healthy closed population rats were used as donors and SD rats as recipients. The Wistar to SD model of graft rejection was developed. All the 42 recipients were randomly divided into 2 groups: group A receiving nothing except CsA (5mg?kg~ -1 ? d~ -1 ) in the first 10 days after operation, and group B receiving MMF (10 mg/kg) and CsA after operation. On the 3rd, 5th and 10th week, all the allografts were tested by light microscopy and immunofluorescence. Pathological changes of the kidneys and the expression of C4d in allograft tissue were observed.Results From the 3rd week, the rats in group A showed light pathological changes of chronic rejection and they became more and more obvious as time increased. Pathological changes occurred in group B at the 5th week and lighter than in group A. At the same time, C4d deposition in PTC was obviously observed on the 3rd week in group A, and on the 10th week C4d widespread deposition in allografts.Conclusion The deposition of complement split product C4d in allografts appears earlier than pathological change of chronic rejection, which can be regarded as a significant indicator to predict chronic rejection of renal allografts.