10.3969/j.issn.2095-4344.2013.26.016

Objective To analyze the characteristics and significance of matrix metalloproteinase-9 (MMP-9) expression in the pathophysiological processes of tuberculous meningitis in mice.Methods Sixteen mice were intracerebroventricularly injected with H37RV suspension as the model group.Meanwhile,the other 16 mice were injected with 0.9％ sodium chloride solution as the control group.Thirty days later,all mice were decapitated and the brain tissue were respectively used to for Mycobacterium tuberculosis (M.tuberculosis) incubation,pathological changes observation,MMP-9 activity detection by zymography,blood-brain-barrier permeability and moisture content detection,and immunofluorescence stain of MMP-9,glial fibrillary acidic protein (GFAP) and integrin αM (OX-42).The t test was used to compare the differences between the two groups.Results Every experimental mouse was injected with (1.271±0.111) × 106 colony-forming units (cfu) M.tuberculosis.Thirty days later,the amount of M.tuberculosis in brain tissue homogenates was (4.900± 1.407) × 104 cfu/mL,and the hematoxylin and eosin staining showed dilatation of subarachnoid and ventricular and infiltration of a large number of inflammatory cells.The cumulative absorbance (A) of MMP-9 bands of brain tissue was 47 821 ± 19 932 in the model group and 10 082 ± 3 544 in the control group.The difference was statistically significant (t =3.728,P=0.010).The evans blue (EB) content of brain tissue was (11.8 ± 3.6) μg/g in model group and (4.7 ±3.4) μg/g in control group.The difference was statistically significant (t=2.887,P=0.028).The moisture of brain tissue was 0.849±0.035 in model group and 0.775±0.037 in control group.The difference was statistically significant (t=2.925,P=0.026).The immunofluorescence staining showed that the infected brain tissue expressed high degrees of MMP-9,GFAP and OX-42.And MMP-9 was overlapped with both GFAP and OX-42 obviously.Conclusions The activity of MMP-9 is significantly enhanced in brain tissue of mice suffering from tuberculous meningitis and participates in blood-brain barrier damage,tissue edema and inflammatory cells exudation.Microglia cells-astrocytes network is involved in the secretion of MMP-9.

Object Through performing the service satisfaction survey,to learn their interest claim and discover the service defects ,so as to provide advice and countermeasures.Method Using field survey methods and statistics analysis to study.Result The satisfaction of relatives of pediatric patients at the level of "relatively good" and "good" is about 60% both in medical treatment environment and equipment ,over 70% in medical treatment effect,doctor capacity,nursing capacity and overall assessment,but only 5.1% in medical expenses.Conclusion Relatives of pediatric patients have high satisfaction in treatment capability of medical staff,medical treatment effect as well as overall assessment ,normal satisfaction in medical environment and equipment,but lowest satisfaction in medical expenses,and the satisfaction in tertiary hospital is not high.Relatives in some districts demand much in some aspects,such as improving hospital environment,attaching importance to ward hygiene ,improving service attitude,and providing single ward ect.

Objective To evaluate the in vitro antifungal activity of Cinnamaldehyde in combination with Voriconazole (VRC) against clinically isolated Aspergillus fumigatus strains. Methods According to the Clinical and Laboratory Stan?dards Institute (CLSI) M38-A2 document,the minimal inhibitory concentrations (MIC) of Cinnamaldehyde and VRC alone or in combination against 42 clinical Aspergillus fumigatus isolates were determined by both microdillution method and check?board method respectively. The MIC50, MIC90, MICG and MICs distribution were compared between single drug and both in combination. The concentration-accumulative curve was drawn and fractional inhibitory concentration index (FICI) was cal?culated to evaluate the interaction between two test agents. Results The values of MIC50, MIC90 and MICG were significant?ly decreased (P<0.001) when combination of the two drugs than those of their single use, with their MIC distribution concen?tration-accumulative curves shifted to the left. The value of FICI of Cinnamaldehye-VRC combination ranged from 0.187 5 to 1.5. Sixteen strains (38.10%) of them showed the synergistic effect, 19 strains (45.23%) showed additive effect, and 7 strains (16.67%) showed an unrelated effect, and no antagonist effect on tested Aspergillus fumigatus strains in vitro. Conclu?sion Cinnamaldehye in combination with VRC mainly shows a combined synergic and additive inhibitory effect on Asper?gillus fumigatus isolates, and this combination appears to be more active against the test strains, which are less susceptible to voriconazole.

Objective To investigate the occurrence of postoperative pain of hepatectomy and its possible related factors.Methods The clinical data of 555 cases undergoing hepatectomy was analyzed retrospectively,and the related influencing factors on postoperative pain of hepatectomy were analyzed by univariate analysis and multivariate logistic regression.Results Moderate postoperative pain was reported in 255 cases among 555 patients who underwent hepatic resection (with an incidence of 45.95％).Incision pain which was often sharp was most common,followed by postoperative complication caused pain.According to whether the postoperative pain occurred or not,all cases were divided into postoperative pain group (n =255) and non-postoperative pain group (n =300),univariate analysis showed that age (P ＜0.01),surgical history (P ＜ 0.01),surgical approach (P ＜ 0.01),incision length (P ＜ 0.01),xiphoid removal(P ＜ 0.01),the final outcome of incision (P ＜ 0.01),complications (P ＜ 0.01) were significantly different between the two groups.Logistic multiple regression analysis showed that the independent influencing factors of postoperative pain included surgical history (P =0.001),surgical approach (P =0.005),incision length (P =0.000),xiphoid process removal (P =0.001),complications (P =0.000).Conclusions The postoperative pain of hepatectomy has a high incidence.Surgical history,surgical approach,incision length,xiphoid process,removal and postoperative complications are the independent impact factors of postoperative pain.

Objective To discuss the treatment of periprosthetic femoral fractures (PFF) with unstable prosthesis by replacement of long-stem femoral prosthesis and internal fixation.Methods From December 2005 to December 2014,15 PFF patients with unstable prosthesis (15 hips) following were treated at our department.They were 10 men and 5 women,aged from 64 to 89 years (mean,76.2 years).Their primary surgeries included total hip arthroplasty in 13 cases and biological bi-polar replacement of femoral head in 2.Two prostheses were cement and 13 biological.By Vancouver classification,9 cases were type B2,and 6 type B3.The unstable prostheses in the 15 cases were replaced by long-stem femoral ones,followed by internal fixation.At the last follow-ups,clinical outcome were evaluated by Harris scoring and images of PFF by Beals & Tower criteria.Complications were documented.Results One died 4 months after operation.The other 14 patients were followed up for an average of 4.5 years (from 6 months to 9 years).Fracture union was achieved in 12 cases after an average of 3.9 months (from 3 to 9 months).Nonunion occurred in 2 cases.Imaging evaluation revealed 9 excellent cases,3 good ones and 2 poor ones.The Harris scores at the last follow-up averaged was 82.3 points (from 50 to 100 points).Deep vein thrombosis occurred preoperatively in one case and posterior tibial vein thrombosis occurred in 2 cases respectively on day 3 and day 10 postoperatively.No such complications occurred as malunion,fixation failure,dislocation or prosthesis loosening.Conclusion Satisfactory outcomes can be achieved by replacement of long-stem femoral prosthesis combined with appropriate fixation for treatment of PFF with unstabrosthesis.

Objective To study the treatment effect of cinnamaldehyde on Aspergillus fumigatus biofilm(BF) in vitro .Meth‐ods The models of A .fumigatus BF were established in vitro ;the minimum inhibitory concentration(MIC) on A .fumigatus was measured .The crystal violet assay and scanning electron microscopy were employed to determined the treatment effect of A .fumiga‐tus biofilm under varying concentrations of cinnamaldehyde .Results BF models were established successfully in vitro .MIC value of A .fumigatus of cinnamaldehyde was 256 μg/mL ;The biofilm biomass in serially increasing concentrations of cinnamaldehyde(1 MIC ,1/2 MIC ,1/4 MIC)were 0 .81 ± 0 .11 ,1 .13 ± 0 .18 and 1 .59 ± 0 .11 respectively .Compared to untreated control group(2 .18 ± 0 .15) ,difference achieved statistical significance(P<0 .05) .SEM studies revealed the deformity of three‐dimensional structures of biofilms treated with sub‐MICs of cinnamaldehyde .Conclusion Cinnamaldehyde has significant antifungal activity against Aspergil‐lus fumigatus ,sub‐MICs could disrupt the mature biofilm in vitro .

BACKGROUND: Our previous findings indicate that inflammation-activated bone marrow mesenchymal stem cell conditioned medium (MSC-CM) contribute to repairing the structure and function of the small intestine after radiation-induced acute intestinal injury. However, it is unclear whether the repair effect can be achieved by regulating small intestinal stem cells. OBJECTIVE: To investigate the effects of inflammation-activated bone marrow MSC-CM on the small intestinal epithelial stem cells after acute radiation-induced intestinal injury and to further discuss the repairing mechanism. METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats were separated, cultured and identified. Then, the bone marrow mesenchymal stem cells were co-cultured with normal or radiation-induced IEC-6 cell lines in the Transwell system for 24 hours. Inflammation-activated bone marrow mesenchymal stem cells were cultured alone for 48 hours. Non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR) were collected. Adult Sprague-Dawley rats (provided by the Experimental Center of Sun Yat-Sen University North Campus) were randomly divided into four groups with 20 rats in each group: control group, radiation group, radiation+MSC-CMNOR group and radiation+MSC-CMIR group. The rats in the latter three groups were exposed to one-off 14 Gy whole abdominal radiation to make a rat model of acute radiation-induced small intestinal injury. Three-day continuous administration beginning within 4 hours after successful modeling was given via the tail vein and intraperitoneal implantation of Alzet micro-osmotic pumps: EMEM-F12 (200 μL/d) for the radiation group, MSC-CMNOR for radiation+MSC-CMNOR group and MSC-CMIR for radiation+MSC-CMIR group. There was 2 mL of concentrated conditioned medium in the pump which was released at a constant rate of 10 μL/h into the abdominal cavity after implantation. Intestinal samples were collected at 1, 3, 5, 7 days after radiation for immunochemistry staining, western blot and qRT-PCR detection. RESULTS AND CONCLUSION: (1) On the 3rd day after radiation, Lgr5 positive cells, which were actively proliferating on the base of crypts, became significantly reduced compared with the normal control group, and there was nearly no existing Lgr5 positive cells. However, after infusion of MSC-CMIR, Lgr5 positive intestinal stem cells were significantly increased compared with the radiation group, while in the radiation+MSCNOR group, there was no significant increase in Lgr5 positive intestinal stem cells. (2) On the 3rd day after radiation injury, Bmi1 positive intestinal stem cells were almost invisible. After infusion of MSC-CMIR, Bmi1 positive intestinal stem cells increased significantly, and it was observed not only in the +4 cell position but also in the common position used to be Lgr5 stem cells, indicating that Bmi1 stem cells could differentiate into Lgr5 positive cells to act its repairing effect. (3) Western blot and qRT-PCR further confirmed that the radiation+MSC-CMIR group was significantly higher on the Lgr5 expression level than the radiation group and the radiation+MSC-CMNOR group, and it returned to the normal level on the 7th day after the continuous high expression level. The repair effect of radiation+MSC-CMNOR group was weaker, and only on the 7th day, the expression level of Lgr5 was statistically different from the radiation group. To conclude, inflammation-activated bone marrow MSC-CM exert a protective effect on the small intestinal epithelial stem cells after acute radiation-induced intestinal injury

OBJECTIVETo explore the clinical significance of high-sensitivity C-reactive protein (hsCRP) in the development of chronic hepatitis B (CHB).

METHODSA total of 182 patients with untreated CHB and 50 healthy individuals (controls) participated in the study. Correlation analysis was performed to determine the association of serum hs-CRP with the age,sex,medical history,serum hepatitis B virus (HBV) DNA, liver function parameters,liver stiffness measure (LSM) and hepatic fibrosis; in addition, correlation analysis was carried out for the associations of degree of liver damage with grade of hepatic fibrosis, LSM and the serum levels of hs-CRP.

RESULTSCHB patients showed significantly higher serum hs-CRP levels than healthy controls (2.38 ± 2.79 vs.0.78 ± 1.07; t =2.495, P < 0.05). Serum hs-CRP levels were significantly correlated with HBV DNA (r = 0.159), liver function parameters (total bilirubin, r = 0.271; alanine aminotransferase, r = 0.298; aspartate aminotransferase, r = 0.389), and LSM, r = 0.562) (all P < 0.05). The correlations with liver function (r = 0.340), LSM (r = 0.292) and hepatic fibrosis grade were positive (r = 0.434) (all P < 0.01).

CONCLUSIONSerum hs-CRP levels in CHB patients can reflect degree of liver damage and of liver fibrosis.

A mouse-anti-human monoclonal antibody was produced by using the membrane proteins of human lung carcinoma cell line A549 as the immunogen to generate monoclonal antibodies against lung carcinoma with the use of hybridoma techniques. McAb4E7 was prepared successfully. To identify its antigen, proteomic technologies such as two-dimenstional electrophoresis, western blotting and mass spectrometry were employed. The targeting antigen of McAb4E7 expressed positive in human lung cancer cell lines A549 and human hepatocarcinoma cell line HepG2, moreover, the expression of the antigen was stronger in A549 cells. Finally, we obtained one positive protein in A549 cell line that has strong affinity and specificity for McAb4E7, which was identified to be ATP synthase beta subunit. We identified ATP synthase beta subunit as the targeting antigen of lung carcinoma special monoclonal antibody McAb4E7.
Animals ; Antibodies, Monoclonal ; biosynthesis ; chemistry ; immunology ; Antibodies, Neoplasm ; immunology ; Antibody Specificity ; Antigens, Neoplasm ; genetics ; immunology ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; immunology ; Membrane Proteins ; immunology ; Mice ; Mice, Inbred BALB C ; Mitochondrial Proton-Translocating ATPases ; immunology