Auditory Cortex ; metabolism ; Humans ; Magnetic Resonance Spectroscopy

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

AlM:To study the treatment effect of traditional Chinese medicine in the treatment of high myopia macular hemorrhage, using Chinese medicine etiology and pathogenesis, syndrome differentiation treatment, and provide the basis for the clinical treatment. METHODS: Eighty patients ( 135 eyes ) with high myopia macular hemorrhage were selected in the hospital from January 2012 to september 2014 as treatment group, and applied traditional Chinese medicine treatment. Forty-five patients (64 eyes) with the same period, as the control group, received routine western medicine treatment. After 1mo treatement, the treatment effect and vision improvement situation of two groups were observed, and after 6mo follow-up, the relapse was observed.RESULTS: The total effective rate of treatment group was 85. 19% (115/135), higher than the control group 78. 13% (50/64) (P<0. 05). The average corrected visual acuity of treatment group was 0. 48±0. 11, higher than the control group 0. 36 ± 0. 09, the difference was statistically significant (P<0. 05). The average diopter and macular bleeding scope of the treatment group were -9. 81±0. 85D and 0. 51 ± 0. 27PD, lower than the control group -10. 76 ± 0. 91D and 0. 78 ± 0. 23PD, the differences were statistically significant ( P < 0. 05 ). The eye ground hemorrhage absorption time of treatment group was 25. 34±2. 28d, less than the control group 29. 72 ± 2. 13d, the difference was statistically significant (P<0. 05). The bleeding again of the control group 7. 81% ( 5/64 ), higher than the treatment group was 5. 19% (7/135), the difference was statistically significant (P<0. 05).CONCLUSlON: Evidence-based treatment of traditional Chinese medicine for high myopia macular hemorrhage has good clinical effect, can shorten the treatment time, and is beneficial to the recovery of postoperative vision, worthy of clinical popularization and application.

Objective To detect the gene mutation of thyroid hormone receptor β ( TRβ ) in a family with thyroid hormone resistance syndrome.Methods The genomic DNA was extracted from peripheral blood leukocytes of the patient and his 5 family members.The exons 1-10 ofTRβ gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.Results Two members of this family were confirmed to have the C y A transition mutation at nucleotide 1642 site within exon 10 of TRβ gene,which was a missense mutation causing the substitution of Proline to Threonine (P453T).The mutation was Heterozygous.Conclusions It was confirmed that the patient has TRβ gene mutation P453T in exon 10.The mutation may lead to the occurrence of thyroid hormone resistance syndrome.

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

BACKGROUND: Pancreas-kidney transplantation is an effective treatment for diabetes combined with final stage renal disease. However, as the patients suffer diabetes for a long period of time, and cardiovascular system disease is complex, pre- and post-transplantation treatment is very important for successful pancreas-kidney transplantation.OBJECTIVE: To discuss immunosuppressant, coagulant, perioperative and postoperative treatment during pancreas-kidney transplantation to provide some clinical experience for pancreas-kidney transplantation.METHODS: Clinical data of 5 cases undergoing simultaneous pancreas-kidney transplantation in Department of Urinary Surgery, the 309 Hospital of Chinese PLA General Hospital between 2003 and 2008 were retrospectively analyzed to summarize the application of immunosuppressants and anticoagulant drugs and perioperative clinical monitoring focus. RESULTS AND CONCLUSION: There were 5 male patients with an average age of 43 years, and suffered type I diabetes mellitus complicated with final stage renal disease. The preoperative insulin dosage was 1.5-2.4 U/(kg·d). One case had diabetic retinopathy and fundus oculi hemorrhage for many times; two cases showed apparent coronary atherosclerotic heart disease with preoperative cardiac ejection fraction of 52% and 50%. Exocrine of transplanted pancreas had been considered by the intestinal fluid drainage. A total of 3 cases were complete rehabilitation. Of them, 1 case developed acute rejection in the first seven days after operation, but renal function restored with the hormones impact; 1 case had postoperative acute rejection of transplanted duodenum as well as intestinal fistula, eventually, transplanted pancreas was ectomized, but transplanted kidney was preserved; two cases succeeded in restoring and no complications occurred; 1 had postoperative gastrointestinal bleeding and died from multiple organ failure. Simultaneous pancreas-kidney transplantation is the most effective way to treat the diabetes mellitus with terminal nephropathy. Because of complications in the transplanted exocrine pancreas with bladder drainage, it has been replaced by the enteric drainage. Recovery of the transplanted kidney function is important for successful transplantation. After operation, oral FK should be taken when the serum creatinine returned to 300 umol/L. The application of clotting drug is one of the important factors for recovery of transplanted pancreatic function. Jejunostomy is an important therapeutic measure to prevent the reflux of intestinal juice to the transplanted pancreas in perioperative period. In the follow-up period cathartic drugs are recommended to prevent constipation and reduce the occurrence of acute pancreatitis caused by intestinal fluid reflux.

To investigate the inhibitory effects of Ginsenoside Rb1 (GRb1) on apoptosis caused by Herpes Simplex Virus-1 (HSV-1) in Human Glioma Cells (U251),U251 cells were infected by HSV-1 at a multiplicity of infection of 5 and GRb1,GRb1+HSV-1,HSV-1 and control groups.MTT and cell apoptosis assays were used to detect the inhibitory effects of GRbl on the apoptosis of U251 cells that caused by HSV-1 infection for various concentrations of drug and virus treatments by MTT assay.We found that in the 400 μg/mL GRbl and 400 μg/mL GRbl+HSV-1 groups,MTT values were higher than control group at all times (P＜0.05).Moreover,the apoptosis rate in the 400 μg/mL GRb1+HSV-1 group was lower than the HSV-1 group (P＜0.05).These results confirmed that,at appropriate concentrations,GRb 1 could inhibit nerve cell apoptosis in HSV-1 infections.

BACKGROUND: Brain death patient is the optimal donator due to the short warm ischemia time, which is conductive to renal function recover following transplantation. However, there are no reports concerning the recovery of renal function in uremia patients following renal transplantation with brain death patients' kidney. OBJECTIVE: To summarize the experience and therapeutic efficacy of renal transplantation using brain death free-donated kidney. METHODS: Six patients with urinsmia underwent renal transplantation with donor kidney of brain death patients from May 2006 to November 2008 at the Organ Transplantation Center, 309~(th) Hospital of Chinese PLA, were selected, including 2 patients receiving kidneys from a brain death patient, 4 patients receiving kidneys from 3 brain death patients. Four recipients received immunosupprsssive regimen of mycophenolate+ciclosporin+steroid, and 2 recipients received mycophenolate+ acrolimus+steroid. The renal function and medicine density were detected regularly, and change of renal function and pathogenetic condition were retrospective analyzed. RESULTS AND CONCLUSION: All 6 patients accepted renal transplantation successfully. The serum creatinine level was obvious descended in 5 patients within a week after transplantation, which meant that the transplanted kidney had begun to work. One patient suffered delayed renal graft function, and returned to normal at 10 days after transplantation. Three patients suffered acute rejection in the first year, and recovered by intravenous glucocorticoid therapy. One patient died after 1 year for pulmonary infection, which accompanied by serum creatinine of 469 pmol/L. The other patients reexamined regularly, and they had good quality of life up to now. The results reveled that renal function recovers in time after transplantation using brain death free-donated kidney, which can ameliorate life quality of patients.

BACKGROUND: Living-donor kidney transplantation is not only associated to prognosis of recipients, but also donors' healthy. Complete medical and psychological assessment should be performed prior to transplantation to ensure the safety of donors. OBJECTIVE: To analyze and summarize the assessment experience of living-relative kidney donors prior to transplantation. METHODS: Totally 77 cases of living-relative kidney donors admitted at Organ Transplantation Center, Second Affiliated Hospital of General Hospital of Chinese PLA between January 2006 and March 2008 were reviewed. Among them, the analysis was carried out respectively according to the live donor nephrectomy guide of the United Kingdom (2005 Edition) before January 20, 2008, and live donor kidney transplantation consensus Boao meeting after January 20, 2008. In common practice, hypertension, diabetes mellitus, cardiovascular system, infectious disease study, age, obesity, proteinuria, renal artery, renal function, receptor for HLA typing and medical ethics, were systematically evaluated. RESULTS AND CONCLUSION: Of the 77 cases of assessed patients, 69 were qualified, successful donors, and completely cured, without complications. Totally 8 cases of non-donors were due to: 2 cases for hypertension combined with end organ damage; 2 for diabetes mellitus; 1 confirmed malignant tumor in kidney-donated surgery; 1 in the activity period of hepatitis B; 1 for resistance from his wife with medical ethics. The average age of donors was 45.3 years old, including and 7 cases above 60 years old, 24 of 50-59 years old, 29 of 30-49 years old, and 9 below 30 years old. There were 39 cases of parent child donation, 1 child parent donation, 19 siblings donation, 7 cases of three generations of collateral serum, and 3 cases of donation between husband and wife. Of successful donors, blood pressure was above 140/90 in 8 cases; Successful donors were without symptoms of myocardial ischemia but 14 cases had consciously ECG ST-T changes; 3 cases had abnormal fasting blood glucose. The successful donors' body mass index (BMI) reached the average of 23.05 kg/m~2, were below 30 kg/m~2; In assessment of infectious diseases, 3 cases were hepatitis B HBs and HBc-positive in a non-activity period, and the antibody titers were below 500 IU/L. In renography, glomerular filtration rate (GFR) was assessed to average (137.3±28.5) mL/min, and no significant statistical difference emerged (P < 0.05). Vascular three-dimensional CT prompted many left renal artery root in 3 cases, accounting for 4% in successful donors, 1 case did not match, accounting for 1%. It demonstrated that the primary purpose of assessment of -living-donor renal transplantation is to ensure that the adaptability, safety and health of donors. In order to avoid the omission of important medical information and unnecessary invasive inspection, as well as reducing the fees of medical assessment, the assessment should be based on the familiar, universally recognized, clinical evidence-based and reasonable procedures and the flexible assessment process.