Humans ; Kidney ; metabolism ; pathology ; Kidney Failure, Chronic ; metabolism ; therapy ; Kidney Transplantation ; Leptin ; metabolism ; Renal Replacement Therapy

Objective To explore the role of miR-184 in Oxygen-Glucose-Deprivation (OGD) induced SK-N-SH cell ischemic injury and its regulation on AKT2 level. Method We used a combination of oxygen and glucose deprivation to imitate ischemic conditions in vivo. MiR-184 mimic and inhibitor were transfected into SK-N-SH cell to alter miR-184 levels. The expression of miR-184 and AKT2 were determined by using Real-time PCR. The extent of SK-N-SH cell survival rate was assessed by thiazolyl blue tetrazolium bromide (MTT) assay. Result Here, we observed that miR-184 was significantly inhibited in SK-N-SH cell after OGD (P<0.05). The changes of miR-184 level altered the expression of AKT2 mRNA. In addition, alteration of miR-184expressionsignificantly affected cell survival rate after OGD. Conclusion miR-184 plays an important role in ischemic injury through negatively regulating AKT2 level, which may provide a potential therapeutic target for ischemic stroke in miRNA levels.

BACKGROUND: Antiviral drugs can reduce the incidence of early-onset cytomegalovirus(CMV)disease,but are associated with strong toxicity and the development of late-onset CMV disease.In order to prevent CMV disease better,cytotoxic T lymphocytes(CTL)may play a critical role in controlling CMV reactivation.Fluorescent HLA-peptide tetramers are used to monitor the recovery of CMV CTL in recipients of allogeneic transplants.OBJECTIVE: To explore the effect of HLA-peptide tetramers and adoptive immunotherapy in treating CMV disease.METHODS: A computer-based online search of Pubmed and Wanfang databases was performed for articles related to CTL detection,application of antiviral drugs and HLA-peptide tetramers,and adoptive immunotherapy with key words"HLA-peptide tetramers,cytomegalovirus,specific CTL,adoptive immunotherapy"in English and Chinese.Repetitive articles were excluded and 29 articles were included.RESULTS AND CONCLUSION: Adoptive immunotherapy with CMVs cytotoxic T cells as preemptive therapy is a very elegant strategy; however,generation of these cells is costly and time-consuming,and therefore the therapy is not available at every transplantation center.Magnetic selection of CMVs CD8+T cells from peripheral blood of CMV-seropositive donors by using HLA-peptide tetramers may be very hopeful,which simplifies adoptive immunotherapy.

Objective To assesses the value of the Pipkin fracture classification on the treatment and prognosis of Pipkin fracture. Methods A total of 71 patients with Pipkin fractures treated from January 2002 to January 2007 were followed up and the detailed information of 63 patients were obtained. The clinical and radiographic evaluation criteria of Thompson was employed to evaluate the outcome, analyze the results and discuss the correlation between Pipkin fracture type and prognosis and hence propose the significance of Pipkin classification on the prognosis. Results There was no statistical difference in aspect of the outcome between type Pipkin Ⅰ , Ⅱ injury and type Pipkin Ⅳ injury (types Pipkin Ⅰ and Ⅱ injury combined with minimum fracture, with fragment ＜ 1 cm,P＞0. 05). There showed statistical difference in outcome between types Pipkin Ⅰ , Ⅱ injury and type Pipkin Ⅳ injury (types Pipkin Ⅰ and Ⅱ injury combined with acetabular rim fracture, P ＜0. 05). Conclusions Pipkin fracture classification system needs a further improvement. The type Pipkin Ⅳ injury that is combined with types Pipkin Ⅰ , Ⅱ , Ⅲ injuries with minimum fracture (fragment ＜ 1 cm) of the acetabular rim should be incorporated into types Pipkin Ⅰ ,Ⅱ , Ⅲ injury. Type Pipkin Ⅳ injury refers to types Pipkin Ⅰ ,Ⅱ , Ⅲ injuries, with major fracture of the acetabular rim and the hip joint instability. In addition, the Pipkin fracture type involving the fracture line, femoral neck and intertrochanteric region is hard to treat clinically and has poor prognosis, suggesting that this type of injury should be considered as special type Pipkin Ⅲ injury.

Objective: To establish a simple,rapid and novel Rayleigh light scattering(RLS)method for rapid determination of mexiletine hydrochloride in drugs. Methods: In the presence of acid Tris-HCl medium and cetylpyri- dinium bromide,eosin Y reacted with mexiletine hydrochloride to form a ternary ion association complex with two charac- teristic scattering peaks by electrostatic attraction. The detection wavelengths were 368 and 586 nm. There was a linear relationship between the mexiletine hydrochloride concentration in a certain range and the Rayleigh light scattering en- hancement intensity(ΔIRLS)of the association complex. Single-wavelength Rayleigh scattering(SWO-RLS)method or dualwavelength Rayleigh light scattering(DWO-RLS)method was used to determine the content of mexiletine hydrochloride, and the mexiletine hydrochloride content was calculated according to the regression equation of standard curve. Results: The linear ranges of mexiletine hydrochloride were 0.005-0.65 mg/L(SWO-RLS method,368 nm),0.004-0.65 mg/L (SWO-RLS method,586 nm)and 0.004-0.65 mg/L(DWO-RLS method,368 nm+586 nm),respectisely. Detection lim- its were 0.0033(SWO-RLS method,368 nm),0.0040(SWO-RLS method,586 nm)and 0.0018 mg/L(DWO-RLS method, 368 nm+586 nm),respectisely. The recovery and relative standard deviation(RSD,n=5)for a SWO-RLS method were 98.6-103% and 1.4-1.8%,respectively(SWO-RLS method,368 nm). Conclusion: The method is simple,rapid, highly sensitive and high selectie.

Animals ; China ; Elapidae ; Humans ; Male ; Snake Bites ; Snake Venoms ; poisoning ; Young Adult

Objective To observe the effects of endoplasmic reticulum stress (ERS) on the activation of monocytes induced by high glucose and explore the underlying mechanism.Methods The monocyte cell line THP-1 was stimulated with high glucose,and then treated with molecular chaperone betaine.The levels of glucose regulation protein 78 (GRP78) and p-JNK,which were associated with ERS were detected by real-time PCR and Western blotting.The proliferation of the cell line was detected by MTT method.Transwell and immunofluorescence were applied to observe the chemotaxis and phenotype of cells respectively.Results The levels of GRP78 and p-JNK of THP-1 cells stimulated by high glucose were significantly increased compared with the normal control group (all P ＜ 0.05).The proliferation and chemotactic were also enhanced (all P ＜ 0.05).The number of cells in M1 phenotype was increased remarkably (P ＜ 0.05).All the indexes above could be rescued by betaine.Conclusion The activation of THP-1 cells can be induced by high glucose through ERS,while molecular chaperone betaine can reverse the activation.

BACKGROUND: The inoculation of hybridoma cell strain onto mouse abdominal cavity may obtain ascites containing mass antibody. Previous method to purify monoclonal antibody in ascites is complex and difficult to operate.OBJECTIVE: To prepare, purify and label anti-human leukocyte antigen (HLA)-I class molecule light chain monoclonal antibody, and to detect the expression of tumor cell surface HLA-I class molecules. METHODS: Hybridoma cells were inoculated onto the mouse abdominal cavity. Ascites containing anti-human light chain beta2-microglobulin antibody were obtained and purified with the modified caprylic acid-ammonium sulfate method. The purified monoclonal antibody was labeled with fluorescein isothiocyanate to detect peripheral blood mononuclear cells, T2 cells expressing blank HLA-A2 molecule and K562 cells surface HLA-I class molecules. The expression of HLA-I class molecules was determined by using flow cytometry and fluorescent microscopy. RESULTS AND CONCLUSION: The purified anti-human light chain beta2-microglobulin-fluorescein isothiocyanate monoclonal antibodies accounted for 96% purity. Flow cytometry results showed that, the HLA-I class molecules were highly expressed in peripheral blood mononuclear cells surface, lowly expressed in T2 cells, and not expressed in K562 cells surface. It is a simple and convenient method to purify ascites with the modified caprylic acid-ammonium sulfate method, and according prepare anti-human light chain beta2-microglobulin-fluorescein isothiocyanate. This method is effective to distinguish the levels of HLA-I class molecules expressed in various cells.

Objective Based on active monitoring MRSA carriage for hospitalized patients, the relationship between colonization pressure and MRSA cross transmission in wards without rigorous contactisolation measures was analyzed, and the role of colonization pressure in predicting MRSA cross transmission was further evaluated. Methods From March to December 2009, active MRSA colonization screening was performed for 240 hospitalized patients in emergency ward and 94 cases in RICU in our hospital. rep-PCR method was employed to do homology analysis on MRSA strains obtained in this study. MRSA weekly colonization pressure, threshold colonization pressure ,cross transmission rate were calculated respectively. RR of MRSA cross transmission under higher level of colonization pressure and lower level of colonization pressure was analyzed. Results MRSA carriage rates on admission for patients in emergency wards and RICU were 6. 25% (15/2A0) and 13. 83 % (13/94) ,and MRSA cross transmission occurred in 13 weeks and 14 weeks in above two units, respectively. Threshold colonization pressures for above two units were 6. 49%and 17. 66%, respectively. For emergency ward, the MRSA cross transmission rate under higher level of colonization pressure was significantly higher than that under lower level of colonization pressure (x2 = 7. 10,P＜0. 01), the RR of MRSA transmission was 9. 61 (95% CI:1. 25-74.00). For RICU, the MRSA cross transmission rate under higher level of colonization pressure was significantly higher than that under lower level of colonization pressure(x2 = 12. 60, P＜0. 01 ), the RR of MRSA transmission was 15.87 (95% CI:2. 06-122. 10). Conclusions Higher level of colonization pressure is an important risk factor for MRSA transmission, and average colonization pressure can be used as a prediction index for MRSA transmission and strengthening prevention and control measures.

Objective To discuss the operative methods, timing and outcomes of severe open Pilon fractures. Methods From April 2003 to July 2008, 21 patients with open Pilon fractures were admitted. All the patients were type C fracture according to AO/OTA classification, including 17 cases of type C2 and 4 cases of type C3. According to Gustilo classification, there were 18 cases of type Ⅱ, 2 cases of type ⅢA, 1 case of type ⅢB. The patients were treated in two different ways: one-stage minimally invasive osteosynthesis for reconstruction of the articular surface with transarticular external fixation of the ankle (19 cases),which included 16 cases of Gustilo Ⅱ AO/OTA C2, 2 cases of type Gustilo Ⅱ AO/OTA C3 and 1 case of type Gustilo ⅢA AO/OTA C2. Formal open reconstruction of the articular surface by plating and external fixation was performed when the condition of soft tissue had improved (2 cases), which included 1 cases of type Gustilo ⅢA AO/OTA C3 and 1 cases of type Gustilo Ⅲ B AO/OTA C3. Results All the patients were followed up from 6 to 48 months, with the mean of 24 months. The outcome of reduction was evaluated by the Burwell-Charnley score. Anatomical reduction was found in 6 cases, function reduction was in 14 cases and poor reduction was 1 case. The heal-time ranged from 2.5 to 11 months, with the mean of 4.7 months. The delayed union occurred in 2 cases. The rate of delayed union was 9.5%. Two patients experienced skin superficial necrosis. There were superficial infection in 2 cases and deep infection in 1 case. The infection rate was 14.3%. Eight cases experienced early traumatic arthritis. The incidence of this complication was 38.1%.According to American Orthopedic Foot Ankle Society (AOFAS) scale for ankle joint, the result of the treatment was 66-94, with an average of 85.2. Conclusion It is important to have the optimal timing of surgery and reliable fixation according to fracture classification, the condition of soft tissue and time after injury. It is the key to evaluate the condition of soft tissue and protect the blood supply.