Pseudolaricis Cortex and its adulterants were identified using DNA barcoding technique in this study. The sequences of ITS2 of Pseudolaricis Cortex and its five adulterants including 37 samples were analyzed. Se-quences were assembled using CodonCode Aligner 3.5.7. K2P distances were calculated and NJ tree was per-formed applying MEGA5.0. The ITS2 secondary structure was predicted using the ITS2 database and website. The results showed that the haplotypes of ITS2 regions of Pseudolaricis Cortex were the same as that of the original plant leaves. The inter-specific K2P distances of ITS2 were obviously higher than the intra-specific ones. Sam-ples of Pseudolaricis Cortex were clustered into a single clade in the NJ tree. The secondary structure of ITS2 of Pseudolaricis Cortex was significantly different from its adulterants. Therefore, ITS2 could powerfully discriminate Pseudolaricis Cortex and its adulterants.

Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.

Objective To study the effects of extracellular-signal regulated kinase mitogenactivated protein kinase (ERK-MAPK) signaling pathway inhibition on histone phosphorylation and the related gene expression in human colorectal cancer cells.Methods Two human colorectal cancer cell lines (SW1116 and HCT116) were cultured and treated with gradient(0,20,40/μmol/L) doses of ERK-MAPK signaling pathway inhibitor U0126.Cell viability was determined by cell counting kit 8 (CCK-8) assay.Cell cycle distribution was assessed by flow cytometry.The expression levels of histone H3 kinases including ribosomal S6 serine-threonine kinase (RSK-2) and mitogen-and stressactivated protein kinase 1 and 2 (MSK1 and MSK2),and the levels of histone H3 (Ser10) phosphorylation and c-Fos protein were detected using Western blotting.Results Treatment of these two human colorectal cancer cell lines with ERK-MAPK inhibitor resulted in a time and dose-dependent inhibition of cell proliferation significantly. Proliferation rate of HCT116 was reduced to 47% at 72 hours after 40/μmol/L U0126 treatment. Cell cycle analysis showed that the percentage of phase G0/G1 cells significantly increased (P<0. 01) and the percentage of phase S cells decreased (P<0.01) after treatment with ERK-MAPK inhibitor. The expression of MSK1 and RSK2 reduced obviously in both of human colorectal cancer cell lines treated with U0126, which resulted in a 28% and 40% reduction of levels of MSK1 and RSK2 as compared with control HCT116 cells respectively,while no detectable change in the expression of MSK2 was found. Consistent with this, the expression level of histone H3 (ser10) phosphorylation was markedly down-regulated by ERK-MAPK inhibitor, and the related protein c-Fos expression decreased accordantly. Conclusions Decreased ERK-MAPK signaling pathway may reduce histone H3 (Ser10) phosphorylation via suppression of the activity of histone H3 kinase including MSK1 and RSK2, but not MSK2, consequently decrease the expression of c-Fos protein, which results in the inhibition of colorectal cancer cells proliferation.

Objective To investigate the effect of ADAR2 on the expression of MAVS and its mechanism.MethodsThe RT-qRCR was used to detect the expression of ADAR gene family, the expression level of ADAR2 and MAVS in cells.The effect of ADAR2 on the 3'UTR region of MAVS was detected by Pyrosequencing.To detect the expression of luciferase by dual luciferase reporter plasmid assay;The expression of ADAR2 and MAVS were detected by Western blot.Results In the ADAR family, the abundance of ADAR1 was the highest, followed by ADAR2, but the expression of ADAR3 was poor, which was almost impossible to detect(P<0.05).ADAR2 played a critical role in RNA editing of chr20:3869744 sites on the 3'UTR region of MAVS(P<0.001).On the 3'UTR editing site of MAVS, the luciferase activity of the edited G allele was significantly lower than that of the normal A allele(P<0.01).At the level of transcription and translation, ADAR2 significantly inhibited the expression of MAVS(P<0.05).Conclusions ADAR2 by editing MAVS` 3'UTR on the chr20:3869744 site, which makes the occurrence of A to G replacement, inhibits the expression of MAVS.

Objective To study the association of multi-drug resistance with mutations in mar gene in clinical isolates of Shigella.Methods Fifty-four clinical isolates of Shigella were collected.Susceptibility tests of tetracycline(TE),chloramphenieol(C),ampicillin(AM),cipr ofloxacin(CIP),and norfloxacin(NOR)were performed in a11 isolates.Hexane and cyclohexane were used to study the organic solvent tolerance of all clinical isolates.marOR genes of these strains were analyzed by potymerase chain reaction-single strand conformation polymorphism(PCR-SSCP)and were sequenced. Results Thirty-five multi-drug resistant strains were identified,and the rate of multi-drug resistance was 64.8%.Thirty-eight of 54 strains were tolerant to organic solvent,including 33 multi-drug resistance strains,3 drug resistant strains and 2 sensitive strains. marOR gene was found in all strains by PCR, and 5 multi-drug resistant strains,(14.29%)carrying marOR gene mutations in multi-drug resistant strains were found by SSCP analysis.DNA sequence analysis of marOR gene revealed that there was a 4 base deletion from base 1376 to 1379 in marO gene that resulted in frame-shift mutation.However,this deletion mutation didn;t exist in the standard strain S51250 and sensitive strains.There were 10 point mutations of marR gene in multi-drug resistant strains,2 of them resulting in amino acid changes:1752(G→A)Gly→ser,1854(T→c)Tyr→His.The rest mutations were all nonsense,and some of them occurred in sensitive strains or in many other strains.Conclusion The marO gene mutation may play an important role in the regulation of multi-drug resistance in Shigella spp.

A growing number of people with unilateral breast cancer have chosen to undergo contralateral prophylactic mastectomy in order to prevent the occurrence of second primary breast cancer and thus avoid associated treatment and death. Contralateral prophylactic mastectomy has been considered beneficial in high-risk populations, such as premenopausal BRCA1/2 mutation carriers. As a result, contralateral prophylactic mastectomy acceptance in patients with no such mutation is controversial. Contralateral prophylactic mastectomy can reduce the risk by up to 95%. Therefore, it is very important to evaluate the risk of contralateral breast cancer and to make appropriate surgical treatment. A review of risk factors for contralateral breast cancer and the benefits of contralateral prophylactic mastectomy in patients with unilateral breast cancer is presented.

Objective To analyze the effect of eukaryotic expression vector containing sense and antisense DNA methyltransferase (DNMT1) gene on the transcript level of tumor associated genes in human colon cancer cell line. Methods Recombinant plasmid, including sense DNMT1 (HMT) and antisense DNMT1 (THM) gene, were constructed and transfected into SW1116 cell by using the lipofectamine. Then G418 filtration was performed. The expression of DNMT1 protein was examined by Western blotting. The transcription level of hMLH1, hMSH2, c myc and p16 INK4A genes were detected by RT PCR. Results The sense and antisense eukaryotic expression vectors were successfully constructed and then the constructed recombinant plasmids were transfected into SW1116 cell. The protein levels of DNMT1 have been up regulated and down regulated in SW1116 cells transfected with HMT and THM plasmids, respectively. The mRNA level of hMLH1, hMSH2, c myc gene were down regulated in the sense DNMT1 transfected cell. The mRNA level of hMSH2 was up regulated in the antisense DNMT1 transfected cell. However, the transcription level of p16 INK4A gene could not be associated with DNMT1 in SW1116 cell.Conclusion DNMT1 can regulate the expression of the tumor associated genes in human colon cancer cell line SW1116.

Objective To evaluate the clinical effect of proximal femoral nail anti-rotation Ⅱ (PFNA Ⅱ) internal fixation in the treatment of intertrochanteric fracture.Methods From September 2012 to April 2016,the clinical data of 37 patients with intertrochanteric fracture who were treated by PFNA Ⅱ internal fixation in Yuehua Hospital were retrospectively analyzed.Results All 37 patients were successfully operated,and the operation time was 40 ～120 min,with an average of 66 min.The amount of blood loss was 30 ～ 110 mL,with an average of 50 mL.During operation and after operation,X ray showed a good position and steady internal fixation.Three days after operation,patients exercised diseased side hip joint.Seven days after operation,patients exercised sitting on the bed.Fourteen days after operation,suture was dismantled and patients could get out of bed and walk with no weight or some weight.Lower limb deep venous thrombosis (DVT) was not observed.Through operative multiple reexamination,the fracture was healed and hip joint function was good.Conclusion PFNA Ⅱ has advantages of convenient operation,less trauma,stronger internal fixation,rapid recovery.It is an ideal method to treat intertrochanteric fracture.

Objective To access evaluate the efficacy and safety of on-line hemodiafiltration for treating a-cute cerebrovascular disease (ACVD) patients complicated with hyperosmolar nonketotie diabetic coma (HNDC).Method Totally 11 patients of ACVD complicated with HNDC were observed for this prospective control study.All of them underwent on-line hemodiafiltrafion for 90 minutes using Fressnius 4008S Hemodialysis Machine withONLILNEplus TM and F60 one hour after final diagnosis was made. The bicarbonate ultrafiltrate rate was set at 500ml/min and blood flow at 150 ～ 180 ml/min; and the substitute fluid were infused with post-dilution at 50 ～ 60ml/min. The symptom of brain edema and cardiac insufficiency during the same course of treatment was observed.Blood were taken from the patients to detect serum kalium, serum glucose, serum natrium, BUN and plasma os-motic pressure at one hour before treatment and six hours after treatment, respectively. The change of in conscious-ness and adverse effects were evaluated at 24 hours after treatment. Results All patients were treated successful-ly. The blood glucose, serum natrium, serum kalium, BUN and plasma osmotic pressure were decreased after on-line hemodiafiltration. Among the 11 patients, consciousness was improved obviously in 8 patients, 3 patientsdied, accounting for 73% of successful effectiveness. Conclusions The on-line hemodiafiltration was effectiveand safe for treating ACVD patients complicated with HNDC.

Objective To compare the efficacy of continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD) on polycystic kidney disease (PKD) patients with end-stage renal disease (ESRD). Methods Retrospective analysis was made on 29 patients with PKD who carded out dialysis therapy for over 3 months in our department from January 2001 to December 2007. They were divided into the CAPD group (10 cases, 34.5%) and HD group (19 cases, 65.5%). Ten cases of non-PKD CAPD patients were randomly selected as the control, who matched the CAPD group in terms of age and gender. The patient information was recorded, such as general data, initial dialysis data, comphcations, survival time, quit of dialysis or death, etc. Kaplan-Meier method and Log-rank test were adopted to analyze the survival rate. Results The survival rates of 1-, 3- and 5-year for the CAPD group were 90%, 75% and 25% respectively, while for the HD group were 94.4%, 67.6%, and 48.3%, and for the control were 83.3%, 44.4% and 22.2% respectively, with no significant differences among 3 groups (P>0.05). group and the control were quite similar. The incidence of peritonitis for the CAPD group (0.62 times/patient year) was similar to that for the control (0.30 times/patient year)(P>0.05). The duration of the lust peritonitis[(23.5±4.0) months vs (20.0±15.8) months] and the catheter exit-site infection (0 time vs 1 time) for two groups were similar as well (P>0.05). One patient had hernia in CAPD group and no patient in control group had hernia. The incidence of peritoneal dialysate leakage was similar between these two groups. In the HD group, two patients (10.5%) had cerebral hemorrhage resulting in death, and 10 patients (52.6%) had cystic hemorrhage, 5 out of whom underwent operation due to repeated cystic hemorrhage and 2 cases received unilateral nephrectomy because of severe hemorrhage. No patient in CAPD group had cerebral hemorrhage but 1 patient (10%) had cystic hemorrhage and recovered after conservative treatment. The hemorrhage complication incidence of CAPD group was significantly lower than that of HD group (P<0.05). Conclusions The prognosis and complication incidence in PKD and non-PKD patients treated with CAPD are similar. The prognosis of PKD patients treated with CAPD or HD is also similar, and the risk of hemorrhage complications of PKD patients treated with CAPD may be decreased compared with those treated with HD. PKD patients can choose HD or PD as the initial therapy of ESRD unless existence of hernia or intolerance. PKD is not the contraindication of PD.