AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Objective To establish the normal parameters of psychometric measures such as the number connection tests A(NCT-A)and digit symbol tests(DST)in assessment of minimal hepatic en- cephalopathy(MHE).Methods One hundred and sixty healthy volunteers(aged 25 to 64 years;educa- tional level＞9 years)were divided into＜35 ys,35～44 ys,45～54 ys and 55～64 ys groups.All of the healthy volunteers were assessed with NCT-A and DST to establish the normal value of age-related parameters,which can be used for diagnosis of MHE in patients with liver cirrhosis.Two standard devi- ation of the normal mean was used as a diagnostic criterion for MHE.One hundred and six cirrhotic patients were assessed with these parameters.Results The parameters of NCT-A were(25.1?4.6) sec in＜35 ys group,(32.1?6.8) sec in 35～44 ys group,(38.6?7.1)sec in 45～54 ys group or (49.3?6.3)sec in 55～64 ys group.The scores of DST were 49.9?4.7 in＜35 ys group,44.6?4.8 in 35～44 ys group,38.5?5.0 in 45～54 ys group or 35.4?4.7 in 55～64 ys group.Thirty one out of 106 cirrhotic patients were diagnosed as MHE based on these parameters.Conclusion The NCT- A and DST are psychometric assessments for diagnosis of MHE.Age-based normal paramerters of NCT- A and DST are needed to be established.

OBJECTIVETo explore the differential proteomic expression in human liver cells L-02 induced by different dosages of trichloroethylene (TCE).

METHODSHuman liver cells L-02 were treated with different concentrations of TCE and the solvent control (dimethylsulfoxide). The total cellular proteins were separated using 2DE and visualized with silver staining after TCE treatment. The images were analyzed with Image Master 2D Platinum 5.0 analysis software. The differentially expressed protein spots were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS).

RESULTSFifteen protein spots with significant difference were found, and went upward or downward or disappeared after the stimulation of TCE with different dosages, which indicated that TCE induced the change of the proteomic expression in the liver cells. The mass spectrum identification and the IPI human database retrieval were used for identifying 9 proteins related to the L-02 Liver cells induced by TCE.

CONCLUSIONThe result provides an insight to TCE-related molecular mechanism and which might be useful for further study of the TCE-associated proteins and molecular markers.

Cell Line ; Dose-Response Relationship, Drug ; Electrophoresis, Gel, Two-Dimensional ; Hepatocytes ; drug effects ; metabolism ; Humans ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trichloroethylene ; toxicity

BACKGROUNDTuberculosis remains the leading cause of human death. Currently, Bacillus Calmette-Guérin (BCG) is the only available vaccine against tuberculosis but its efficacy is highly variable. Thus, developing new tuberculosis vaccines becomes an urgent task. In this study, we evaluated in BALB/c mice the humoral and cellular immune responses of recombinant BCG expressing the antigen ESAT-6 from Mycobacterium tuberculosis.

METHODSEscherichia coli-BCG shuttle plasmid named pDE22-esat-6 was constructed by inserting the BamHI/EcoRI digested esat-6 gene PCR product into the similarly digested parental plasmid pDE22. BCG cells were transformed with pDE22-esat-6, which was named recombinant BCG (rBCG). BALB/c mice were immunized subcutaneously on the back with 100 microl normal saline containing 10(6) CFU of BCG or rBCG. They were sacrificed after 4 weeks to detect their humoral and cellular responses.

RESULTSThere was no any significant differences in the growth characteristics between the conventional BCG and rBCG. In immunized mice, the IgG antibody titres of rBCG group were as high as 1:8000, which was significantly higher than that in BCG group (1:1400, P < 0.05). The elicited IFN-gamma level of rBCG group was (1993 +/- 106) pg/ml, which was also significantly higher than that in BCG group ((1463 +/- 105) pg/ml, P < 0.05). The splenocyte proliferation index of rBCG group reached 4.34 +/- 0.31, which was higher than that of BCG group (3.79 +/- 0.24, P < 0.05).

CONCLUSIONrBCG secreted expressing antigen ESAT-6 stimulated stronger humoral and cellular immune responses than BCG did, and, therefore may be the better vaccine against mycobacterium tuberculosis.

Animals ; Antigens, Bacterial ; genetics ; immunology ; BCG Vaccine ; immunology ; Bacterial Proteins ; genetics ; immunology ; Interferon-gamma ; biosynthesis ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Recombinant Proteins ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo analysis the early complications of tibial fracture and its related factors, and propose a solution.

METHODSFrom December 2003 to December 2013,38 patients with early complications of tibial plateau fracture after operation were retrospectively analyzed. There were 35 males and 3 females, aged from 37 to 69 years old (averaged 42.3 years). According to Schatzker classification, 3 cases were classified as type II, 2 cases as type III, 2 cases as type IV, 19 cases as type V, 12 cases as type VI. The intervals between injury and operation ranged from 9 hours to 9 days, 26 cases within 3 days. Fifteen cases were treated with internal fixation of plates and 23 were treated by plate fixation and bone transplantation. Early complications included skin necrosis in 15 cases, infection in 6 cases, osteofascial compartment syndrome in 3 cases, common peroneal nerve injury in 2 cases, the superficial peroneal nerve injury in 3 cases, popliteal artery injury in 2 cases, loss of reduction in 7 cases.

RESULTSThe wound of 14 cases healed at the first stage and 24 cases healed delay. Hospitalization days ranged from 7 to 67 days (averaged 25.6 days). All patients were followed up for 12 to 36 months with an average of 16.4 months. The fracture healing time ranged from 3 to 9 months (averaged 6.9 months). According to Merchant knee function evaluation criteria, the results were excellent in 19 cases, good in 12, fair in 5 and poor in 2.

CONCLUSIONEarly complications of tibial fracture after operation is closely associated with the severe fracture complexity and related with preoperative preparation, surgical timing, operation incision selection and surgical technique. Early detection and timely processing reduce damage.

Adult ; Aged ; Female ; Humans ; Length of Stay ; Male ; Middle Aged ; Postoperative Complications ; therapy ; Tibial Fractures ; surgery

OBJECTIVETo study the relationship between blood lactate level and disease severity in critically ill children.

METHODSThe clinical data of 232 children who were critically ill between September and December, 2010 were retrospectively studied. According to blood lactate levels within 24 hrs of admission, the 232 patients were classified into three groups: normal (n=146), high lacticemia (n=72) and lactic acidosis (n=14). The circulation functions, pediatric critical illness scores and prognosis were compared among the three groups.

RESULTSThe degree of sepsis among the three groups was different (χ2 = 13.592, P<0.01). The occurrence of septic shock in the lactic acidosis group (42.9%) was significant compared with that in the normal (7.5%) and the high lacticemia groups (11.1%). The pediatric critical illness scores were different among the three groups (χ2 = 12.854, P<0.05). The blood lactate level was significantly negatively correlated with the pediatric critical illness scores (r=-0.405, P=0.002). The prognosis among the three groups was also varied (χ2 = 25.599, P<0.01). The curative rate (7.1% vs 23.3%; P<0.05) and the improvement rate (28.6% vs 58.2%; P<0.05) in the lactic acidosis group were significantly lower than in the normal group, and the mortality (28.6%) was significantly higher than in the normal (5.5%) and the high lacticemia groups (6.9%).

CONCLUSIONSA higher blood lactic acid level is associated with a more severe illness state and a worse prognosis.

Adolescent ; Child ; Child, Preschool ; Critical Illness ; Female ; Humans ; Infant ; Lactic Acid ; blood ; Male ; Prognosis ; Retrospective Studies ; Severity of Illness Index

One case of pneumoscrotum associated with spontaneous colon perforation was reported. The patient was a 66-year-old man, presented with high temperature, mild abdominal pain and an enlarged scrotum. Physical examination revealed scrotal swelling, abdominal tenderness Case Report and muscular defense. Computed tomography (CT) of the abdomen showed swelling and pneumatosis of the left major psoas and iliopsoas muscles, and ultrasound found subcutaneous emphysema of the scrotum. Surgical investigation discovered a retroperitoneal perforation in the descending colon connected with a huge retroperitoneal vomica and scrotal sac. Spontaneous colon perforation induced pneumoscrotum is rare clinically. It may present as colon perforation, which calls for special attention.
Aged ; Colonic Diseases ; complications ; Genital Diseases, Male ; etiology ; Humans ; Intestinal Perforation ; complications ; Male ; Scrotum ; Subcutaneous Emphysema ; etiology

OBJECTIVETo investigate the treatment for patients with early stage primary tonsil non-Hodgkin's lymphoma (NHL).

METHODSTwo hundred and thirteen patients with previously untreated early stage primary tonsil NHL were reviewed. All patients were pathologically confirmed. According to Ann Arbor classification, 35 patients were stage I, 178 stage II. The primary treatment for stage I was radiotherapy alone in 12 and combined modality therapy (CMT) in 23 patients. The primary treatment for stage II was radiotherapy alone in 57,chemotherapy alone in 2, and CMT in 119 patients.

RESULTSThe 5-year overall survival, cancer specific survival (CSS) AND disease-free survival (DFS) for the early stage primary tonsil non-Hodgkin's lymphoma were 65%, 70% and 61%, respectively. The 5-year CSS was 63% for the radiotherapy alone group and 72% for the CMT group (p = 0.064), and the 5-year DFS were 56% for the radiotherapy alone group and 62% for the CMT group. For patients with stage I disease, The 5-year CSS were 100% in both radiotherapy alone and CMT groups, and the 5-year DFS were 100% and 80% in these two groups (p = 0.148), respectively. There was no significant difference of efficacy between the two treatment s for the patients with stage I disease. For the patients with stage II disease, the 5-year CSS was 58% in radiotherapy alone group and 66% in CMT group (p = 0.051). However, CMT significantly improved DFS in stage II disease, with a 5-year DFS of 46% for radiotherapy alone and 60% for CMT (P = 0.046).

CONCLUSIONPatients with stage I tonsil non-Hodgkin's lymphoma treated with radiotherapy alone or CMT can achieve an excellent outcome. CMT significantly improve the DFS in stage II patients. There was a trend that CMT improved the survival rates in the patient with early stage disease. It was suggested that CMT should be used for the patients with early stage primary tonsil non-Hodgkin's lymphoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Combined Modality Therapy ; Female ; Humans ; Lymphoma, Non-Hodgkin ; mortality ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Retrospective Studies ; Survival Rate ; Tonsillar Neoplasms ; mortality ; pathology ; therapy

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Cell Cycle Proteins ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Minichromosome Maintenance Complex Component 2 ; Nuclear Proteins ; metabolism