Objective To investigate the changes of expression of apoptosis-related proteins Bcl-2,Bax and Fas of the transplantated hepatocytes after intraperitoneal transplantation in rats using porcine hepatocytes cultured with polylactic acid-O-carboxymethylated chitosan nanoparticles.Methods Recirculating collagenase perfusion method was used to isolate porcine hepatocytes.At 48h after D-galactosamine intraperitoneal injection,5 mL polylactic acid-O-carboxymethylated chitosan nanoparticles-cultured porcine hepatocytes embedded in collagen gel(group A) and 5 mL porcine hepatocytes embedded in collagen gel(group B) were transplanted into peritoneal cavity of SD rats,respectively.The amount of transplanted porcine hepatocytes was 5.0?107 hepatocytes in two groups.The expression of Bcl-2,Bax and Fas in transplanted hepatocytes three days after transplantation was detected with immunohistochemistry.Results At 1,2 and 3d after transplantation,the rates of Bcl-2 positive cells in group A was significant higher than those in group B,whereas the rates of Bax and Fas positive cells in group A were significant lower than those in group B(P

BACKGROUND:Previous research has proved good therapeutic effect on the acute hepatic failure by the treatment of polylactic acid-O-carboxymethylated chitosan(PLA-O-CMC) nanoparticles-cultured porcine hepatocytes embedded in type I collagen.OBJECTIVE:To investigate the effect of PLA-O-CMC nanoparticles on immunological rejection in intraperitoneal porcine hepatocyte xenotransplantation for treatment of acute liver failure rats.DESIGN,TIME AND SETTING:Randomized control animal experiments were carried out at the Institute of Hepatobiliary Surgery and Laboratory of Nerve Regeneration in Nantong University from May 2005 to May 2006.MATERIALS:In situ recirculating collagenase perfusion method was used to isolate porcine hepatocytes.64 SD rats were intraperitoneally injected with D-galactosamine to induce acute hepatic failure models.METHODS:Totally 64 rats were divided into four groups:Model group received no intervention;Nano-collagen hepatocytes group was transplanted with PLA-O-CMC nanoparticles-cultured porcine hepatocytes embedded in collagen gel,which was then wrapped up using greater omentum;Porcine hepatocytes embedded in collagen gel(collagen hepatocytes group) and free porcine hepatocytes suspension(pure hepatocytes group) were transplanted into peritoneal cavity of SD rats,respectively.At 48 hours following the modeling,the porcine hepatocytes transplantations were performed and the number of transplanted cells was 5.0?107 hepatocytes in all groups.MAIN OUTCOME MEASURES:The serum interleukin-2,interferon-?,IgG and IgM levels of rats were determined at days 1,2,3,5 and 7.The pathological changes of transplants were observed under microscope at days 1,3 and 7.RESULTS:There were no significant differences in serum interleukin-2 and interferon-? levels of all groups during 7 days post-transplantation.Serum IgG levels increased with a peak at day 3 after hepatocytes transplantation,and then began to decline.Serum IgG levels in nano-collagen hepatocytes group were lower than those in collagen hepatocytes group and pure hepatocytes group at days 2 and 3(P

Objective To investigate immunological relationship between early induction of food allergy and occurrence of later asthma in mice, and explore the pathological changes in lung tissues. Methods Thirty-seven female BALB/c mice aged 6 weeks were randomly divided into blank control group ( n = 12), food allergy group ( n = 13) and asthma group (n = 12). After being challenged by ovalbumin (OVA), the levels of serum IgE, IL-4 and INF-γ in bronchoalveolar lavage fluid ( BALF) were detected. The numbers of inflammatory cells and eosinophils ( EOS) in BALF were counted. Lung tissues were obtained for pathological sections, and thickness of bronchial wall and EOS infiltration were observed. Results The level of serum IgE and level of IL-4, ratio of IL-4/IFN-γ and number of EOS in BALF in food allergy group and asthma group were significantly higher than those in control group (P <0.05). The level of IL-4 and number of EOS in BALF in asthma group were significantly higher than those in food allergy group (P <0.05) , while there was no significant difference in serum IgE level between these two groups (P > 0.05), and levels of IFN-7 in BALF in both groups were significantly lower than that in blank control group (P<0.05). There were more EOS infiltration in lung tissues and thicker bronchial wall in food allergy group and asthma group than that in blank control group (P < 0.05), and the number of EOS in asthma group was significantly higher than that in food allergy group ( P < 0.05). Conclusion IgE-mediated immune response is involved in both food allergy and asthma mouse models. Lung immune imbalance of Thl/Th2 and inflammatory cell infiltration caused by food allergy may participate in the occurrence of later asthma.

Objective To study the pathogens and drug resistance profiles of pulmonary infection in patients with AIDS. Methods The pathogens and their drug susceptibility of pulmonary infection diagnosed by fibrobronchescopy-induced brunchoalveolar lavage fluid (BAI.F) culture and/or transbronchial biopsy in 116 AIDS cases were analyzed. Results Monopathogenic infection in lungs were detected in 18 cases(15.5%) and mixed infection in 98 cases ( 84.5%). Of the 116 cases, bacteria were present in 91 patients, fungi in 62, tubercle bacillus in 49, pneumocystis jiroveci in 29, and cytomegalovirus in 11.Ninety-five bacterial strains were isolated from BALF, mainly including Streptococci (34), coagulase negative Staphylococcus (20), Klebsiella pneumoniae (10) and Escherichia (7). The isolated bacteria were resistant to β-lactam, macrolides, quinolones and aminoglycosides, of which were 14 methicillin-resistant Streptococci (MRS) strains and 12 extended spectrum β-lactamases (ESBL) strains. Sixty-eight fungal strains were isolated, including 36 Candida mycodermas, 19 Penicilliums, 6 Aspergilli and 5 Mold fungi;they were sensitive to amphotericin B but resistant to fluconazol (5.6% -50. 0% ) and itraconazole( 10. 5%-60. 0% ). Conclusion Pneumonia in AIDS patients are usually caused by multiple pathogens,predominantly consisting of multiresistant bacteria and fungi. Therefore, antibiotics should be rationally chosen according to drug susceptibility test.

AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages.
METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups.
RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.

Objective To observe the resection of the scapholunate interosseous ligament (SLIL)and its subregions on the three-dimensional(3-D)movement of the scaphoid and lunate so as to discuss the role of SLIL in the 3D flexion-extension motion of the scaphoid and lunate.Methods Twelve upper extremities(six left extremities and six right extremities)from adult cadaver were used in this study and divided into five groups:normal group,proximal subregion resection group,proximal subregion plus dorsal subregion resection group,proximal subregion plus palmar subregion resection group and whole SLIL resection group.The 3-D laser scan and reconstruction technique were used for meusure ment of the 3-D flexion-extension motion of the scaphoid and lunate.Results In the normal group,the scaphoid and lunate flexed and the radial deviated at the same direction during wrist flexion-extension motion.At the same time,there was minimal scaphoid and lunate pronation-supination during wrist flexionextension.After resection of the proximal and palmar(or dorsal)subregions of the SLIL,some different movements were found compared with the normal specimen.Whole SLIL resection resulted in increase of the flexion motion of the scaphoid but decrease of the palmar flexion of the lunate.Conclusions 3-D laser scanning and image reconstruction techniques can accurately measure the 3D motion of the scaphoid and lunate.Partial or whole resection of SLIL may exert significant effect on the flexion-extension motion of the scaphoid and lunate.However,the proximal subregion of SLIL has no prominent effect on the motion of the scaphoid and lunate.

ObjectivesTo investigate the clinical value of the melting curve analysis-based PCR assay for the clinical genetic diagnosis and prenatal diagnosis of β-thalassemia.Methods A total of 451 peripheral blood samples,including 372 cases with β-thalassemia phenotypes and 79 cases without β-thalassemia phenotypes,were collected by Liuzhou Municipal Maternity and Child Healthcare Hospital between January 2011 and August 2011.Moreover,another 84 cases,including 16 fetal villi samples (10 - 13 weeks),64 amniotic fluid samples (16 -24 weeks ) and 4 umbilical cord blood samples (above 17 weeks),whose parents were β-thalassemia carriers,were also collected for this assay between June 2011 and September 2011.A double-blind test was done to compare the detection reliability of the melting curve analysis-based assay (24 β-thalassemia mutations can be detected) with PCR-RDB probe assay (17 β-thalassemia mutations can be detected ) and DNA sequencing using these samples.The wildtype,mutant and total concordance rates of the genotyping results were calculated separately among the melting curve analysis based assay,PCR-RDB probe assay and DNA sequencing.Results Among the 451 peripheral blood samples,thirteen mutations and nineteen genotypes were obtained by using melting curve analysis-based assay.447 samples had the same detection results and 4 samples had different detection results by comparing melting curve analysis-based assay with PCR-RDB probe assay,thus,the concordance rate of the sample detection result was 99.1％ (447/451),and the concordance rate of the allele detection result was 99.6％ (898/902).DNA sequencing results of the 4 samples showed that 3 samples had the same genotyping result with melting curve analysis-based assay,and 1 sample had the same genotyping result with PCR-RDB probe assay.A rare β-globin mutation which was not included by melting curve analysis-based assay was not detected.Thus,the genotypes of 450 samples were detected accurately by melting curve analysis-based assay,and the concordance rate of the sample detection between the melting curve assay and DNA sequencing assay was 99.8％ (450/451).Among 84 fetal villi,amniotic fluid,and umbilical cord blood samples,8 mutation types and 18 genotypes were obtained by using melting curve analysis-based assay.All the samples have the same detection results by comparing melting curve analysis-based assay with PCR-RDB probe assay and DNA sequencing,so the concordance rate of the genotyping results was 100％ among the melting curve analysis-based assay,PCR-RDB probe assay and DNA sequencing.Conclusions The melting curve analysis-based PCR assay can detect multiple unknown samples simultaneously,and detect multiple mutations accurately.It is very useful for the genetic diagnosis and prenatal diagnosis of β-thalassemia.

Objective To investigate the antibacterial effect of grape peel residue procyanidins on common clinical bacteria ,inclu‐ding E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,Enterococcus and methicillin‐resistant Staphylococcus aureus (M R‐SA) .Methods By the means of agar plate method ,we detected the minimum inhibitory concentration (MIC) of 30 strains of E .co‐li ,28 strains of Pseudomonas aeruginosa ,15 strains of Acinetobacter baumannii ,32 strains of Feces Enterococcus ,25 strains of E . faecalis and 70 strains of MRSA ,then calculate the MIC50 and MIC90 .The result was analyzed with SPSS software 16 .0 .Results The concentration of procyanidins at 3 -8 mg/mL had no inhibitory effect on E .coli ,Pseudomonas aeruginosa and Acinetobacter baumannii;there was no inhibitory effect on E .faecalis at 3-8 mg/mL ,but the inhibition rate to Feces Enterococcus was 6 .3% .By measuring the inhibitory effect of procyanidins on 70 MRSA ,the inhibition rate at 4 mg/mL was 65 .7% ,at 8 mg/mL was 95 .7% , MIC50 was 4 .221 mg/mL and MIC90 was 6 .260 mg/mL .Conclusion There are no inhibitory effects of grape peel residue procya‐nidins on Gram‐negative bacilli such as E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,and there are certain inhibitory effects on Gram‐positive bacteria such as MRSA ,enterococci ,especially on MRSA (P<0 .05) .

Objective :To explore the feasibility and clinical value of real time three‐dimensional transesophageal echo‐cardiography (RT‐3D TEE) in diagnosis of congenital heart valvular disease (CHVD).Methods :A total of 135 CH‐VD patients treated in our hospital were selected .All patients received surgery ,and transthoracic echocardiography (TTE) and RT‐3D TEE inspection successively within 7d before surgery .Heart valve lesion condition was observed , and diagnostic results of two methods and surgical outcome were compared and analyzed .Results :RT‐3D TEE could display the morphological structure ,lesion degree and peripheral blood flow of heart valves in CHVD patients in a multi‐angle ,stereoscopic and clear way .It could find heart valve disease which is difficult to be identified by TTE , and corrected the diagnostic deviation .With surgical results as the gold standard ,diagnostic coincidence rate of RT‐3D TEE was significantly higher than that of TTE (97. 04% vs.91. 11%, P＝0.039).CHVD diagnosed by RT‐3D TEE and TTE possessed a intermediate consistency (Kappa＝0.477 , P＝0. 001).Conclusion :RT‐3D TEE can pro‐vide more imaging information for the diagnosis of CHVD ,which can be used as an effective supplement for preop‐erative TTE examination .

OBJECTIVETo observe the clinical effects of combined type external fixator in treating complex tibial metaphyseal fractures.

METHODSFrom January 2007 to July 2012, 34 patients with complex tibial metaphyseal fractures were treated with combined type external fixator in different stagings. There were 23 males and 11 females, with a mean age of 41.3 years (ranged, 16 to 63), and the course of disease were from 1 h to 8 d. In the patients, 31 cases were open fractures, 11 cases with type II, 13 cases with type III A, 7 cases with type III B according with Gustilo classification; 19 cases were tibia plateau fractures, 6 cases with type II, 1 case with type IV, 5 cases with type V, 7 cases with type VI according to Schatzker classification; 15 cases were distal tibial fractures (one were bilateral fractures), 2 fractures with type A2, 1 fracture with type A3, 1 fracture with type C1, 5 fractures with type C2, 7 fractures with type C3 according to AO classification. Rasmussensn scoring system and AOFAS Ankle Hind-foot Scale were respectively used to assess the joint function of knee and hip.

RESULTSWound surface of 19 patients obtained at phase I healing and 15 patients obtained at phase III healing. Superficial wound infections occurred in 2 cases and bone non-union necessitated reoperation occurred in 2 cases (final fractures obtained bone healing after the second operation). All patients were followed up from 6 to 38 months with a mean of 14.3 months. At the final follow-up,according to Rasmussensn scoring system, 5 fractures got excellent results, 11 good, 3 fair, the mean Rasmussen score was 23.58 +/- 3.98; according to AOFAS Ankle Hind-foot Scale, 5 fractures got excellent results, 8 good, 3 fair, the mean AOFAS Ankle Hind -foot Scale was 80.75 +/- 14.21.

CONCLUSIONCombined type external fixator can well maintain the stability of the fractures, had advantages of low incidences of soft tissue complications and less influence to joint motion in treatment of complicated tibial metaphyseal fractures. However there were some limitations in long-term use.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Fracture Fixation ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Radiography ; Recovery of Function ; Tibial Fractures ; diagnostic imaging ; physiopathology ; surgery ; Treatment Outcome ; Young Adult