Suppression subtractive hybridization(SSH),which is feasible and highly specific,is one of the most efficient methods for cloning differentially expressed genes.It has been successfully applied in the study of tumors,stem cells,etc.

Objective To investigate the clinical effect of high dose of calcitriol on end-stage renal disease (ESRD) in patients with secondary hyperparathyroidism (SHPT). Methods The clinical data of patients with ESRDcomplicated with SHPT were retrospectively analyzed. According to the treatment method were divided into control group and observation group, the control group were given conventional dose of calcitriol treatment, the observation group were treated with high dose of calcitriol. The differences of calcium and phosphorus levels, parathyroid hormone (PTH) level and quality of life were observed between the two groups before and after treatment. Results The serum phosphorus, calcium and calcium-phosphorus product in two groups of patients before treatment had no significant difference. After treatment, the above indicators decreased in two groups of patients compared with those before treatment, and the observation group was significantly lower (P<0.05). Before treatment, there was no difference in the levels of PTH between the two groups. After treatment, the level of PTH in the observation group was lower than that in the control group, and the difference was statistically significant (P<0.05). There was no significant difference in quality of life between the two groups before treatment. After treatment, the quality of life in the two groups were higher than before treatment, and the observation group was significantly higher (P<0.05). Conclusion High dose of calcitriol has good therapeutic effect for ESRD patients with SHPT and can significantly improve the patient's calcium and phosphorus and PTH levels, improve the quality of life of patients.

Objective To observe the effect of hybrid renal replacement therapy (sustained lowefficiency hemodiafiltration) in recipients with delayed graft function (DGF) after kidney transplantation.Methods In 33 kidney transplant patients with DGF,there were 15 cases subject to sustained low-efficiency hemodiafiltration (SLEDF group),18 cases subject to hemodialysis (HD group).The renal function was tested,and the expression levels of complements C3,C4,and CRP,IL-1β,IL-6,TNF-α were detected before and after therapy.Results In SLEDF group,the renal function recovered in (29 ± 13) days after therapy,and in (47 ± 21) days in HD group.After therapy,the BUN and Scr in both two groups were lower than before (P＜0.01).After therapy,the levels of IL-1β,IL-6 and TNF-α were also lower than before in two groups,and the levels were lower in SLEDF group than in HD group (P＜0.05).The levels of complements C3,C4,and CRP in two groups were decreased after therapy,but there was no statistically significant difference (P＞0.05).Conclusion SLEDF can decrease the level of SCr,BUN and some cytokines in recipients effectively,and It's helpful to recovery of the renal function.SLEDF can be used by DGF recipients in transit time.

Objective To observe the clinical therapeutic effect of nucha electroacupuncture on patients with adult hypoxic ischemic encephalopathy(HIE)and to approach its mechanism. Methods After exclusion of cases with incomplete historical data from 80 adult HIE patients admitted in the Department of Critical Care Medicine of 401st Hospital of Jinan Military Region of PLA,finally 74 cases were randomly divided into a nucha electroacupuncture treatment group(38 cases)and a conventional therapy control group(36 cases)by random number table. The patients in the control group were treated with conventional medical therapies,including dehydration,intracranial pressure reduction,brain protection with head hypothermia,and so on. In nucha electroacupuncture group,based on the conventional treatment in the control group,additionally nucha electroacupuncture was performed for the patients. Before and after therapy,the changes of Glasgow coma scale score(GCS score)and Glasgow-Pittsburgh coma score (G-PCS score)in all the patients were assessed and compared between the two groups,and according to the standard criteria of GCS and G-PCS scores for evaluation of efficacy,the clinical therapeutic effective rates in the two groups were assessed. Results ①There were no statistically significant differences between the two groups in GCS score and G-PCS score before treatment(both P>0.05),after treatment,the GCS and G-PCS scores in nucha electrical acupuncture group were obviously higher than those before treatment(GCS score:6.22±2.66 vs. 4.33±1.35,G-PCS score:22.96±6.22 vs. 17.53±4.68,both P<0.05),and the GCS and G-PCS scores of nucha electroacupuncture group were markedly higher than those of control group after treatment(GCS score:6.22±2.66 vs. 5.17±3.01,G-PCS score:22.96±6.22 vs. 16.78±7.96,both P<0.05).②The total effective rate of nucha electroacupuncture group was significantly higher than that of control group when assessed by both the standard criteria of GCS score(73.7%vs. 50.0%,P<0.05)and G-PCS score(84.2%vs. 61.1%,P<0.05). Conclusion The nucha electroacupuncture has definite clinical therapeutic value in treatment of patients with adult HIE and it can improve their neurological function and outcome.

Objective To explore the effects of cotransplantation with osteoblasts on hematopoietic reconstitution in mice after bone marrow transplantation (BMT). Methods The typical model of syngeneic BMT was established. 18 Balb/c mice were used to prepare the bone marrow nuclear cells and osteoblasts for BMT. The 42 Balb/c mice were randomly divided into 3 group:normal group (6 mice, without any treatment), the single BMT group ( 18 mice, given 2 × 106 bone marrow nuclear cells/each mouse) and the cotransplantation group of HSC with osteoblaats (18 mice,given 2 × 106 bone marrow nuclear cells and osteoblasts/each mouse). The following factors were measured on day 7, 14, 21 after BMT: peripheral blood cells, bone marrow mononuclear cells (BMMNC), the percentage of CD34+ cells in BMMNC (assayed by flow cytometry), the hematopoietic tissue changes (detected by HPIAS-1000 image analysis system) and micro vascular density (MVD) of bone marrow tissue (with immunohistochemistry). Results The levels of periphral WBC, RBC, PLT, BMMNC in the contransplantation group were higher than those in the single BMT group (P＜0. 01 or P＜0. 05). In the contransplantation group, the percentage of CD34+ cells in BMMNC, the hematopoietic tissue area and the MVD of bone marrow were also higher than the single BMT group on the 7th, 14th, 21st day after BMT(P＜0.01 or P＜0.05). Conclusion Cotransplantation with osteoblasts could significantly promote hematopoietic reconstruction in mice after BMT. Cotransplantation may represent a promising means of achieving higher engraftment rate after BMT.

Objective To explore the effects of osteoblasts on the recovery of hematopoiesis and angiogenesis in acute irradiation injury mice.Methods The femurs of 18 male BALB/c mice were used to prepare the bone marrow osteoblasts, and the rest mice were divided into 3 groups as normal group, saline group and osteoblast group.The mice in normal group received no treatment, and the other two groups were received 6.0 Gy 60Co γ-ray irradiation.After irradiation each mouse of osteoblast group was administered with 2 × 106 osteoblasts through tail vein injection, and equal volume saline was given to each mouse of saline group by the same way.The following factors were measured at 7, 14, 21 d after irradiation, they were the counts of peripheral blood cells and bone marrow mononuclear cells ( BMMNC ) , the percentage of CD34 + cells in BMMNC, the histology changes and micro vascular density (MVD) of bone marrow tissue.Results The counts of peripheral blood cells, BMMNC and hematopoietic tissue area in osteoblast group were higher than those in saline group.The percentage of CD34 + cells in BMMNC and the MVD of bone marrow in osteoblast group were also higher than those in saline group at 7, 14, 21 d after irradiation ( t = 2.46 - 64.51, P ＜ 0.05 ).Conclusions Osteoblasts could significantly promote the recovery of hematopoiesis and angiogenesis in mice after acute irradiation injury.

Objective To analyze the diagnosis and treatment of iliac pseudoaneurysms following renal transplantation.Method The data of two patients with pseudoaneurysm who underwent kidney transplantation were retrospective analysied.Result One case with pseudoaneurysm received transplant nephrectomy,and pathological examination diagnosed mucoraceae infection; the other patient received endovascular treatment and amphotericin B therapy,endly died of hemorrhagic shock.Conclusion The development of pseudoaneurysms at the kidney transplantation recipients with the same donor results in high rates of mucormycosis.They should take anti-fungus therapy and operation as early as possible.

Objective To investigate N-terminal pro-B-type natriuretic peptide (NT-proBNP) cutoff value for the mortality in different age groups in critically ill patients.Methods A retrospective study was conducted.295 patients admitted to the intensive care unit (ICU) of 401st Hospital of PLA from January 2011 to October 2012 were divided into two groups according to age [group with age＜65 years old (n=105) and group with age≥ 65 years old (n =190)].The concentrations of serum NT-proBNP,hematocrit (HCT),procalcitonin (PCT),C-reactive protein (CRP),serum creatinine (SCr),estimated glomerular filtration rate (eGFR),acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and probability of survival (PS) were recorded within 24 hours.The primary outcome was ICU mortality.Receiver operator characteristic curve (ROC curve) was used to evaluate the value of NT-proBNP for predicting the mortality.Results ① There were no significant differences in the length of stay in ICU,mechanical ventilation rate,the mortality,the incidence of cardiovascular disease,digestive disease,neurologic disease,and the number of patients having received operation,HCT,PCT and CRP between the two groups (all P＞0.05).The percentage of the male,the APACHE Ⅱ score,the percentage of respiratory disease,and NT-proBNP in group with age ≥ 65 years old were higher than those of the group with age ＜ 65 years old [the percentage of the male:51.6％ vs.33.0％,x2=9.093,P=0.003; APACHE Ⅱ score:22.94 ±8.10 vs.19.44 ±8.51,Z=-3.259,P=0.001; the percentage of respiratory disease:29.47％ vs.17.14％,x2=5.472,P=0.024; NT-proBNP(ng/L):5 859.00(2 050.75,23 802.75) vs.2 882.00 (275.15,6 236.00),Z=-5.514,P=0.000]; PS,the percentage of patients having multiple injuries and other diseases and eGFR in group with age ≥65 years old were lower than those of the group with age ＜65 years old [PS:59.0 (31.5,79.0)％ vs.70.0 (40.0,84.0),Z=-3.431,P=0.001; the percentage of multiple injuries:0.53％ vs.17.14％,x2=30.987,P=0.000; the percentage of other disease:5.79％ vs.13.33％,x2=4.962,P=0.030; eGFR (ml·min-1· 1.73 m-2):81.07 (45.77,131.80) vs.95.54 (33.64,165.55),Z=-2.214,P=0.027].② The area under the ROC curve (AUC) [95％ confidence interval (95％ CI)] of NT-proBNP in patients with age＜65 years old was significantly higher than that of group with age≥65 years old and the entire group [0.825(0.738-0.892) vs.0.664 (0.592-0.731) and 0.725 (0.670-0.775),Z1 =-2.835,P1 =0.005; Z2=-1.995,P2=0.046].③ The sensitivity (76.]0％ vs.64.10％),specificity (82.35％ vs.67.12％),positive predictive value (90.0％ vs.75.8％),and negative predictive value (62.2％ vs.53.8％) with cutoff value of NT-proBNP (2 882 ng/L) in group with age ＜65 years old were significantly higher than those with NT-proBNP cutoff value (6 062 ng/L) in group with age ≥ 65 years old.Conclusion NT-proBNP cutoff value in different age groups for the prediction of mortahty in the critically ill patients maybe more objective and accurate.

BACKGROUND: Presently, acute rejection following renal transplantation remains a risk factor for chronic rejection and graft function injury, How to non-invasive, rapid and exact diagnosis and prompt treatment is important. OBJECTIVE: To investigate early diagnosis and post-treatment expression of urine monocyte chemoattractant protein-1 (MCP-1) in the acute rejection after renal transplantation, through detecting the association of the urine MCP-1 variation according to some cases of nephridial tissue biopsy. METHODS: We selected 62 chronic renal failure patients who received renal homotransplantations in the Department of Renal Transplantation of Zhengzhou People's Hospital from October 2008 to February 2009. The stable renal function group contained 42 patients with stable renal function following renal transplantation. Acute rejection group contained 20 patients with acute rejection following renal transplantation. We chose 10 patients who examined no abnormalities in the Medical Examination Center of Zhengzhou People's Hospital to detect their urine sample as control group. All patients following renal transplantation underwent conventional immunosuppression. In addition, patients in the acute rejection group were treated with antilymphocyte globulin or methylprednisolone reinforced impact therapy. MCP-1 mass concentration changes were measured by double antibodies sandwich enzyme linked immurosorbent assay. RESULTS AND CONCLUSION: Compared with control group, no significant change was determined in urine MCP-1 mass concentration in the stable renal function group (P > 0.05). The urine MCP-1 mass concentration was significantly increased in the acute rejection group (P< 0.01). Compared with pretreatment, urine MCP-1 mass concentration was significantly decreased following treatment in 20 patients from the acute rejection group (P < 0.01). Of them, 17 cases had relieved clinical symptom, and normal auxiliary examination, and their urine MCP-1 mass concentration was close to the control group; 3 cases were inefficient, whose urine MCP-1 mass concentration was greater than the control group. Eight cases received nephridial tissue biopsy, and kidney pathology demonstrated acute rejection of transplanted kidney, which was similar to urine MCP-1 mass concentration in the acute rejection group prior to treatment (P > 0.05). These indicated that the level of MCP-1 in urine can non-invasively diagnose acute rejection following renal transplantation in an early phase, and monitor therapeutic efficacy. This may be associated with renal pathological injury during acute rejection following renal transplantation.

Recent studies indicate that immune-associated aplastic anemia (AA) resembles such autoimmune diseases as insulin-dependent diabetes and chronic autoimmune thyroiditis that belong to organ-specific autoimmune diseases. Many independent investigation groups have successfully isolated the pathopoiesis-associated T cell clone causing hematopoiesis failure with a CD4 phenotype from peripheral blood and bone marrow (BM) in AA patients. In the current study, BM CD4(+) T cells were isolated from AA patients and healthy controls with immunomagnetic beads sorting, and proliferation capability, apoptosis features and the impacts of their secreted cytokines on hematopoiesis stem/progenitor cells were compared between them. By (3)H-TdR method, CD4(+) T cells in AA group presented more enhanced proliferative activity. The stimulation index in control group and AA group was 1.47+/-0.24, and 2.51+/-0.34 respectively (P<0.01). After BM CD4(+) T cells were induced by high concentration of CD3 monoclonal antibody for 18 h, evident apoptosis cells could be seen under the electron microscope in both control group and AA group. Flow cytometry revealed that apoptosis rates in the early and late stages of AA group were significantly higher than in control group (P<0.01). Early-stage apoptosis rate in control and AA groups was (6.85+/-1.48)% and (16.98+/-4.40)%, and late-stage apoptosis rate in control group and AA group was (2.65+/-1.57)% and (7.74+/-0.83)%, respectively (P<0.01). The CFU-GM count in AA group and control group was (74.50+/-9.50)/10(4) cells and (124.25+/-19.80)/10(4) cells respectively under an inverted microscope (P<0.01), and the expression levels of CyclinD3 mRNA and protein in cord blood CD34(+) cells were both down-regulated induced by BM CD4(+) T cell culture supernatant in AA patients. These results indicate that BM CD4(+) T cells of AA patients are likely in an abnormally proliferative, and activated state which can correlate intimately with AA hematopoiesis damage. BM CD4(+) T cells in AA patients can secret some soluble cytokines that can inhibit proliferation of hematopoietic stem cells by suppressing the expression of Cyclin D3, resulting in hematopoiesis failure.