BACKGROUND: Infusion of hemopoietic stem cell from donors can promote the chimeric formation and induce specific immunologic tolerance in the allograft recipients. However, the pretreatment for cell transplantation has great toxicity to recipients. So immunosuppressant combined bone marrow infusion is introduced to anti graft versus host reaction. OBJECTIVE: Based on microchimerism, to study the security and associativity of chimera formation induced by kidney-bone marrow transplantation and immunologic tolerance.DESIGN, TIME AND SETTING: The contrast observation was performed at the department of urinary surgery, The Third People's Hospital of Zhengzhou City from January 1998 to December 2005.PARTICIPANTS: According to ABO/Rh blood type and HLA matching, 96 female patients with chronic renal failure and waiting for kidney transplantation were divided into 2 groups, In the combination group, patients received kidney combined bone marrow transplantation; the other uremia patients received the other kidney of cadavers were served as control group. The donors were 48 healthy males. METHODS: Bone marrow of donors was collected simultaneously with kidney obtain and preserved with cryoprotectant at -198 ℃ in nitrogen canister. After kidney transplantation, large dose of anti-human lymphocyte immune globulin were used for 2 weeks, then (0.9-2.5)×10~8/kg mononuclearcell was reinfused. PCR-SRY was used to identify donor derived cell-chimerism. Lymphocyte subgroup of recipients was determined by blood test; and interleukin 10 was measured by enzyme linked immunosorbent assay; in addition, the mass concentration of tumor necrosis factor α and tumor necrosis factor β was detected. MAIN OUTCOME MEASURES: Chimerism, lymphocyte subsets and cytokines were detected at various time points following transplantation. Simultaneously, the transplantation results and complication status of recipients were observed. RESULTS: The positive rate of chimera in the combination group was greater than that of the control group (P < 0.05). The 3-year follow-up showed that incidence differences of acute rejection between recipients with positive chimera and recipients with negative chimera had significance (13%, 35%, P < 0.05). There was no graft versus host disease occurred in the combination group. CONCLUSION: Kidney-bone marrow transplantation can augment chimerism in early postoperative period, and significantly reduce the rate of acute rejection, which is safe and beneficia1to induce specific immunologic tolerance in the renal allograft recipients.

Objective To report a new approach of splenic hilar lymph nodes dissection in radical gastrectomy for gastric cancer. Methods 193 cases of gastric cancer patient receiving radical resection of gastric cancer between May 2008 and October 2008 were studied. The tail and body of spleen and pancreas were thoroughly freed with retroperitoneal way retrogressively and extruded out of abdominal cavity in 80 cases. The other 113 cases received operation with routine way. Results In 80 cases who received operation with retroperitoneal approach retrogressively, the total splenic hilar lymph nodes were 519, the positive ones were 65, the positive rate was 12.5 %; In the other 113 cases the total splenic hilar lymph nodes were 565, the positive ones were 58, the positive rate was 10.3 %. The positive rate had statistic significance between these two groups. Conclusion Extruded splenic hilar lymph nodes dissection with retroperitoneal approach retrogressively is safe and has the same effect with splenectomy in gastric cancer operation.

BACKGROUND:Studies have shown that malnutrition was associated with death of elder people after fracture of hip, but there were no above-mentioned reports in China.
OBJECTIVE: To retrospectively analyze the relationship of blood albumin, total lymphocyte count and hemoglobin levels to prognosis when elder patients with fracture of hip were on admission.
METHODS:130 elderly patients with hip fractures aged ≥ 70 years were included underwent either total hip arthroplasty or bipolar arthroplasty. Admission serum albumin, total lymphocytecount and hemoglobin levels were recorded. The patients were fol owed up for 1 year or til the death. Survival data were available in 92 patients. Rates of survival were calculated by the Kaplan-Meier method and the Log-Rank test. Cox proportional hazard regression model received prognostic multivariate analysis.
RESULTS AND CONCLUSION:Of the 92 patients, albumin<35 g/L in 20 cases (22%), total lymphocyte count<1.5×106/L in 67 cases (73%), and hemoglobin<120 g/L in 56 cases (61%). Kaplan-Meier method showed that the survival rate of patients with normal albumin (≥ 35 g/L) was significantly higher than those with reduced albumin (<35 g/L) (P<0.01). No significant difference was detected in the survival rate of patients with normal total lymphocyte count (≥ 1.5×106/L) and reduced total lymphocyte count (<1.5×106/L) (P>0.05). The survival rate of patients with normal hemoglobin (≥ 120 g/L) was significantly higher than those with decreased hemoglobin (<120 g/L) (P <0.05). Cox multivariate analysis displayed that albumin decrease is an independent prognostic factor for death of patients with hip fracture. Results indicated that the prognosis of elder patients with hip fracture was strongly associated with their nutritional conditions. Albumin and hemoglobin levels at admission can be considered as important indexes for judging patient’s prognosis.

OBJECTIVE: Strengthen realization of pulmonary infection and perform follow-up plays a key role in preventing severe pulmonary infection following kidney transplantation. The aim of this study is to analyze the diagnosis and pathogens distribution in patients with pulmonary infection following kidney transplantation. METHODS: Totally 14 patients with severe pulmonary infection following kidney transplantation, who received treatment at The Department of Urology Center, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA, from May 2004 to September 2008, were analyzed. Including 10 males and 4 females, aged from 20 to 65 (37.3±11.4) years. Six patients suffered pulmonary infection within 3 months, 7 patients occurred pulmonary infection occurred during 3-6 months, and pulmonary infection occurred in 1 patient at 6 months after kidney transplantation. The infection rate was 93% during 6 months after kidney transplantation. The diagnosis of pulmonary infection was according to the clinical manifestation of respiratory system, and combined with specificity display of sternum, CT or detection of pathogen. Oxygen inhale and mechanical ventilation were employed for admitted patients. And electrolytes, acid-base equilibrium was daily detected. Twelve patients accepted "three-medicine" treatment and 2 cases accepted "four-medicine" treatment. The therapeutic drugs could be regulated individually. The dose of immunosuppressive agent was adjusted due to ciclosporin, CD4~+/CD8~+, and C-reactive protein levels. RESULTS: The pathogens of pulmonary infection were bacteria (5 patients), fungi (4 patients), and mixed infection (3 patients). No pathogen was found in 2 patients. Four patients died of acute respiratory distress syndrome, 1 patient died for quitting treatment. The remained 9 patients were cured. The mortality was 36%, and the recovery rate was 64%, CONCLUSION: The cure rate of patients with severe pulmonary infection following kidney transplantation can be increased by early combining antibiotics treatment, regulating immunosuppressive agents, early identifying pathogens, taking respiratory support therapy, as weii as adding nutritional supplement.

Objective: On the basis of typical cases, the objective of this study is to assess the service quality of primary healthcare institutions under the vertically integrated model from the perspective of demanding side.Methods: The vertical model between Renhe Hospital and Lixian Township Health Center was selected as the case study.Quantitative data were collected by exit review using Primary health-Care quality service Assessment Tool (PCAT)-Chinese version from the patients seeking services in Lixian.Quantitative data were analyzed by descriptive statistics and multiple linear regression equations while qualitative data were analyzed by thematic framework analysis.Results: Under the vertical model, the PCAT total score was 20.09.The scores on the first diagnosis, continuity of care, and comprehensiveness of care and coordination of care were 6.59, 3.27, 4.58 and 5.62 respectively.The scores on the three dimensional extensions, family-centered, community-oriented, medical culture were 3.07, 1.86 and 2.67 respectively.The difference was significant for the PCAT total score between a number of visits and the length of waiting time for services in Lixian.Conclusions: Under the vertical model, primary healthcare institutions did better in the first diagnosis while weaker in continuity and the three dimensional extensions.With the development of the integration, the primary healthcare institutions need to strengthen the continuity and downward referral in the future.Future researches and follow-up, analysis is needed to track the impact of the vertically integrated model.

Objective To study the prevalence and genotype of human parvovirus B19 virus among blood products and plasma pools in China. Methods B19 DNA derived from 16 lots of factor Ⅷ concentrate produced by 4 manufactures and 10 lots of plasma pools were detected by nested PCR. Phylogenetic comparison of the partial B19 sequences obtained from positive samples were performed by direct sequencing. Results Twelve of sixteen lots of factor Ⅷ concentrate and all of ten lots of plasma pools were contaminated by B19 DNA. By comparing the partial B19 sequences,all the isolated viruses were genotype Ⅰ and their nucleotides were high conserved with homology of 98. 3%-100%. Conclusion B19 genotype Ⅰ DNA has been detected in high prevalence in factor Ⅷ concentrate and plasma pools. The genetic diversities were shown to be very low.

Considerable attention has been directed toward studying the impact of diabetes on the central nervous system. The current study investigates the biochemical changes in the brain tissue of streptozotocin (STZ)-induced diabetic rat using 31P magnetic resonance spectroscopy (31P MRS). The 31P NMR spectra of the whole brain show no significant changes of phosphomonoesters and phosphodiesters levels one week after STZ induction, suggesting no apparent structural changes in cell membranes. The results identifies the increased level of adenosine diphosphate, negligible changes of phosphocreatine ( PCr ) and adenosine triphosphate ( ATP) , but the decreased ratio of PCr/ATP, indicating that PCr plays a role of balancing the energy. Moreover, the decreased pH value indicates the changes of the intracellular environment in STZ-diabetic brains in rats. After 15 weeks of STZ injection, the metabolism of phospholipid membrane and brain energy metabolism has been obviously disturbed. Our study successfully shows that 31 P MRS can not only study phospholipid and energy metabolism non-invasively, but also measure intracellular pH and other important biochemical information. All of these spectroscopic characterizations contribute significantly to the understanding of pathogenesis and evolution of diabetes, and provide theoretical basis for early diagnosis and clinical treatment in diabetes.

Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.

The primary importance of magnetic resonance (MR) imaging in evaluating anal fistulas lies in its ability to demonstrate hidden areas of sepsis and secondary extensions in patients with fistula in ano. MR imaging is relatively expensive, so there are many healthcare systems worldwide where access to MR imaging remains restricted. Until recently, computed tomography (CT) has played a limited role in imaging fistula in ano, largely owing to its poor resolution of soft tissue. In this article, the different imaging features of the CT and MRI are compared to demonstrate the relative accuracy of CT fistulography for the preoperative assessment of fistula in ano. CT fistulography and MR imaging have their own advantages for preoperative evaluation of perianal fistula, and can be applied to complement one another when necessary.
Adult ; Anal Canal/anatomy & histology ; Female ; Humans ; *Magnetic Resonance Imaging ; Male ; Middle Aged ; Rectal Fistula/diagnosis/*radiography/surgery ; Tomography, X-Ray Computed ; Young Adult

In order to clone and express alcohol dehydrogenase II (ADH2) gene from Saccharomyces cerevisiae in E. coli BL21 (DE3) efficiently, we extracted the total RNA as template and obtained ADH2 gene by RT-PCR and connected ADH2 gene to pTAT plasmids to gain recombinant expression plasmid pTAT-ADH2, then transformed this recombinant expression plasmid pTAT-ADH2 into E. coli BL21 (DE3). The recombinant was induced by IPTG to express ADH2. After purification, ADH2 activity was tested in vitro and toxicologic test was done in mouse. Sequence test showed that the acquired fragments exhibited 90% homology to ADH2 gene sequence from GenBank report. The target gene expressed efficiently and took up to approximant 50% of total protein by SDS-PAGE and band scanning analysis. The purified protein exhibited the identified activity through biochemical test and mouse toxicological test. As a result, the acquired ADH2 gene was highly homology to the published sequence and expressed at a high level in E. coli BL21 (DE3), more importantly, ADH2 proved to have ethanol dehydrogenase activity.
Alcohol Dehydrogenase ; genetics ; isolation & purification ; Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Mice ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Saccharomyces cerevisiae ; enzymology ; genetics ; Saccharomyces cerevisiae Proteins ; genetics ; isolation & purification