[Objective]To observe the nerve regeneration and functional recovery in the adult rats.[Method]The sciatic nerves on the left side of 15 rats were exposed and 1.0 cm long segments of the nerves were removed from the mid-thigh level and replaced by rabbit nerve made acellular through chemical extraction.At 4 months after procedure,the nerve regeneration and function recovery were examined with HE staining,NF-160 immunohistochemical staining,electrophysiological tests and sciatic functional index(SFI).[Result]In the rats repaired by acellular nerves,regenerated axons re-entered into the acellular xeno-nerve segments without excessive sign of inflammation following implantation.As stimuling(1.5 mA,0.1 ms,1.0 Hz) to the proximal sciatic nerves,the implanted segment resulted in motor evoked potentials,which were recorded from posterior tibial muscles.SFI showed a partial recovery of locomotion of the limb with sciatic nerve defect.[Conclusion]Sciatic nerves defect can be repaired by chemical acellular xenogeneous nerves in rats.There was partial functional recovery,which revealed the grafting with chemical acellular xenogeneous nerves may be a promising mehod for nerve defect in clinical.

The clinical manifestation of acute invasive fungal sinusitis was associated with facial pain,altered sense of smell, blindness and headache. Physical examinations show that dark brown nasal secretions with bone resorption in paranasal sinus. Radiographi parameters showed uneven density in paranasal sinus and intraorbital extension. Fungus smears and pathological examination can make a definitive diagnosis.
Acute Disease ; Facial Pain ; etiology ; Fungi ; isolation & purification ; Headache ; etiology ; Humans ; Mycoses ; complications ; pathology ; Paranasal Sinuses ; Sinusitis ; complications ; microbiology ; pathology

The aim of the present study was to explore the prevalence characteristics of brucellosis in Hainan by investigating the biological characteristics and clinical type of pathogenic bacteria isolated from the blood of a patient infected by infective endocarditis.Bacteriological experiments were conducted according to the standard identification methods of Brucella including morphology,cultural characters,biochemistry characters,serological test and phage test,etc.At the same time,systematic analysis on the information about epidemiology,clinical manifestation and laboratory data of the patient was carried on.The Brucella melitensis was identified as Brucella melitensis biotype 2 and was significantly different from the Brucella melitensis biotypes 1 and 3 isolated in China reported in recent years.The urease test result of Brucella melitensis was variable and there is no strong positive result reported except the isolates in Hainan.As Brucellosis cause by Brucella melitensis biotype 2 was firstly reported in Hainan Province,great importance should be attached to its point-like prevalence.

Objective To express and purify EGF-TCS fusion protein and observe the targeted and selective killing effect on cancer cells of the fusion protein.Methods The recombinant expression plasmid PQE30/EGF-TCS was transformed to E.coli.M15 and the fusion protein(EGF-TCS) was expressed.Ni-NTA Agrose affinity chromatography was used to purify the protein,flow cytometry to detect EGFR expression rate in cancer cells(BEL-7402,MCF-7,BGC-823) and normal liver LO2 cells,and the killing test to verify selective killing ability of EGF-TCS;The cell apoptosis detection by flow cytometry and microscopic observation were used to confirm the selective killing ability of EGF-TCS.Results Recombinant expression plasmid PQE30/EGF-TCS was expressed in E.coli.M15 stably and effectively.The purity of EGF-TCS was over 95% by chromatography.EGFR expression rate was highest in hepatoma cells BEL-7402(72.33%) and lowest in normal liver LO2 cells(5.51%).The killing ability of recombinant protein was more effective to cancer cells(IC50 of BEL-7402,MCF-7 and BGC-823 was 11.4,22.47 and 12.53 ?g/ml respectively) and was weak to normal cells(IC50 53.19 ?g/ml).Conclusion The recombinant protein EGF-TCS that induces apoptosis of cancer cells was successfully constructed by gene engineering technology.

Hemangioendothelioma, Epithelioid ; diagnosis ; Humans ; Temporal Bone ; Vascular Neoplasms ; diagnosis

Objective To deplore the immunoregulatory function changes of mesenchymal stem cells(MSCs)from multiple myeloma(MM)patients and its effects on the pathogenesis of myeloma bone disease.Methods MSCs from MM patients and normal controls were isolated and the immunophenotype was detected.Real-time PCR was performed to detect the expressions of TGF-β1,TGF-β2,TGF-β3,IL-6,IL3,TNF-α,FasL and RANKL of MSCs.Transwell coculture systems were performed between MSCs and T cells.Lymphocyte proliferative assay was employed to detect the effect of MSCs on T cell proliferation.The effect of MSCs on T cell cycle and T cell activation markers CD25 and CD69 expression were analyzed by flow cytometry.Cleaved caspase 3 protein by western blot and hoechst 33258 staining were employed to detect the apotosis of T cells.Influence of T cells on the osteogenesis potential of MSCs were detected by Von kossa stain,real-time PCR and Western blot.Results MSCs from both MM patients and normal controls possessed similar morphology and immunophenotypes.MM derived MSCs exhibited increased expressions of TGF-β1,IL-6,IL-3,TNF-α and RANKL and decreased expression of TGF-β2,TGF-β3 and FasL.The inhibitory effect of MM derived MSCs on T cell proliferative ability was attenuated compared to control MSCs.MSCs from normal controls silence more T cells in Go/G1 phase than those from MM patients.The daupening effect of MM derived MSCs on activation-induced T apoptosis seemed to be enhanced.Expression of T cell activation markers were significantly inhibited by MSCs from normal controls.Both T cells cocultured with MM deprived MSCs and T cells directly from MM patients inhibited osteogenesis potential of MSCs from normal controls.Conclusion MSCs from MM patients showed impaired immunoregulatory capability on T cells.The activated T cells,in turn,inhibited the osteogenesis potential of MSCs.This may participate in the pathogenesis of myeloma bone disease.

Objective To investigate the effect of Bcl-2 gene expression on the proliferation and apoptosis of human pancreatic cancer SW1990 cells.Methods Bcl-2 short guide RNA (Bcl-2-sgRNA) was designed and synthesized,and it was combined with CRISPR-Cas 9.After confirmation by gene sequencing,it was transfected into human pancreatic cancer cell line SW1990,then the cells with stable Bcl-2 gene knock-out were selected,and wild type SW1990 cells were used as control.The cell growth curve was determined by CCK-8 method.The number of clone formation was measured.Flow cytometry was used to measure cell cycle and apoptosis.Results Human pancreatic cancer cell line SW1990 with Bcl-2 gene knock-out was successful constructed.Compared with wild type SW1990 cells,the growth of SW1990 cells with Bcl-2 gene knock-out was inhibited,the number of clone formation was significantly decreased [(160.7 ± 10.0) vs (285.3 ± 14.2)],the proportion of G1 cells was significandy increased [(84.51 ± 0.97) ％ vs (57.49 ± 1.08) ％],the proportion of S phase cells significantly decreased [(12.82 ± 0.99) ％ vs (27.56 ± 1.65) ％],and apoptosis rate was remarkably increased [(12.67 ± 0.59) ％ vs (0.37 ± 0.35) ％],and the difference between the two groups was statistically significant (P ＜ 0.01).Conclusions Knock-out of Bcl-2 gene can inhibit the growth of human pancreatic cancer cell line SW1990,decrease the ability of clone formation,block the cell in G1 phase and greatly increase cell apoptosis rate.

Objective To explore the significance of the plasma procalcitonin (PCT) level for directing antibiotic therapy in elderly patients with ventilator-associated pneumonia (VAP).Methods The 50 elderly patients with VAP were randomly separated into the regular therapy group and the PCT-directed therapy group. The regular therapy group was given regular antibiotic therapy, while the antibiotic therapy was decided according to the plasma level of PCT in the PCT-directed therapy group. The used time and utilization rate of antibiotics, as well as inflammatory indicators including white blood cells, neutrophils, C-reactive protein (CRP) and clinical pulmonary infection score (CPIS) were compared between the two groups. Results After treatment, there were no significant differences in white blood cells, neutrophils and CRP between the PCT-directed therapy group and regular therapy group [(8.9 ± 3.5 ) × 109/L vs. (9.4 ± 3.7) × 109/L, 0.62 ± 0.04 vs.0.60±0.04, (18.7±8.5) mg/Lvs. (21.6±6.0) mg/L, t=0.47, 1.84 and 1.37, allP＞0.05],but the CPIS was markedly lower in PCT-directed therapy group than in regular therapy group [(4.0± 1.4) scores vs. (4.7± 1.0) scores, t= 2. 18, P＜0.05]. The neutrophils, CRP and CPIS were significantly lower after treatment than before in the both groups. The concentration of PCT was decreased after treatment than before [(0.5 ± 0.9) mg/L vs. (1.7 ± 0.7) mg/L]. Meanwhile, the time using antibiotics was longer in regular treatment group than in PCT-directed therapy group [(8.72±1.32) d vs. (5.17±0.72) d, t=11.96, P＜0.01], the utilization rate of antibiotics was higher (95.2 % vs. 55.2 %, χ2 = 12.41, P＜0.01) in regular treatment group. Conclusions Using PCT levels for directing treatment in elderly patients with VAP can achieve better curative effect and reduce the use of antibiotics.

Objective To evaluate the predictive value of brain natriuretic peptide (BNP) on mortality in elderly patient with severe sepsis or septic shock. Method Eighty-three elderly patients meeting with criteria of severe sepsis or septic shock by the American College of Chest Physicians and the Society of Critical Care Medicine (ACCP/SCCM) in 2001, admitted in Intensive Care Unit (ICU) of Zhejiang Hospital during May 2004 to June 2007, were enrolled, and patients with chronic renal failure were excluded. The patients were divided into survivor and non - survivor group according to whether they survived within 28 days staying in ICU, the difference of plasma BNP level,serum C-creative protein (CRP) concentration, APACHE II and SOFA scores on admission day and BNP level on the third day between the two groups were compared; thereafter, the relationship between muhiple variables including age, admission day, BNP, CRP, APACHE 11 and SOFA scores, the 3rd day BNP level and 28-day mortality were analyzed by Logistic regression, and meanwhile the independent predictors for ICU moaality among which were determined. Results BNP levels on both admission day and the 3rd day were significantly higher in nomurvivor group than those in survivor group[ ( 1056.38±676.34) pg/ml vs. (611.59±610.02) pg/ml,p =0.002 and (1448.48+891.11) pg/ml vs. (522.41±575.20) pg/ml, P<0.001, respectively]. By Logistic regression analysis, BNP level on the 3rd day and SOFA score on admission day were independent predictors of ICU mortality, The receiver operating characteristic (ROC) curves indicated that values of areas under the curve of the admission day and the 3rd day BNP levels for 28-days mortality were 0.735 (95% CI,0. 621～0. 848, P <0.001) andO.836 (95% CI, 0.746～0,926,P<0.001), respectively. Conclusions Plasma BNP inereaseds in majority of eldedy patients with severe sepsis or septic shock, which may serve as irdex for prognosis in elderly severe septic paients.

Objective To assess the effects of fluid resuscitation in elderly patients with septic shock by right ventricular end-diastolic volume index (RVEDVI). Method Thirty elderly patients with septic shock with-in 6 hours after onset, admired to intensive care unit of Zhejiang Hospital from January 2007 to October 2008, were randomly divided into control group (n = 15) and experimental group (n = 15). Homodynamic profile of the right ventricular was monitored by using modified Swan-Ganz catheter. Fluid resuscitation was given to the patients of control group monitored by right atria pressure (RAP). The expected efficacy of treatment was the RAP elevated to 8 ～ 12 mmHg. The goal of fluid resuscitation in patients of experimental group was 100～ 200 mL/m~2 RVEDVI corrected by right ventricular ejection fraction (RVEF). RAP, pulmonary artery occlusion pressure (PAOP), RVEF, RVEDVI, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and mixed venous oxygen sat-uration (SvO_2) were recorded before and after the treatment for 6 hours in both groups. The concentration of lactate and the lactate clearance rate of patients in both groups after fluid resuscitation were detected. The relationship between lactate clearance rate and ARVEDVI was evaluated by using correlation analysis. Results The percentage of patients reaching goal of resuscitation in experimental group (86.7%) was higher than that in control group (80%), however, there was no significant difference statistically. In goals-achieving group, RVEDVI, △RVEDVI, RVEF(%), RAP and lactate clearance rate(%) of the patients in experimental group were signifi-cantly higher than those in control group [(119.92 ± 15.65) mL/m~2, (38.54 ± 6.63) mL/m~2, (36.08 ± 3.40), (14.46±1.13) mmHg,(58.31 ± 13.36) vs. (99.92±11.71) mL/m~2,(21.00±11.01) mL/m~2,(32.42± 2.47),(13.08±1.08) mmHg,(43.99±16.26); P <0.05]. However, there was no significant difference in PAOP, APACHE Ⅱ and SvO_2 between two groups (P >0.05). The lactate clearance rate in goals-achieving pa-tients with septic shock has a significant correlation with RVEDVI and △RVEDVI (P < 0.01). Conclusions Fluid resuscitation guided by RVEDVI in elderly patients with septic shock is safe and more effective than that guided by RAP.