The present study was designed to investigate the effects of cytochalasin E(CE) on the integrity of Sertoli cell barrier. The results indicate that: (1) In the CE-treated testis(1000-2000 ?mol/L CE/testis, 6-14 hr), actin filaments of the ectoplasmic specialization (ES) in area of Sertoli cell barrier were disrupted, the accumulation of amorphous material and fragmented small vesicles of SER were observed in cytoplasm of Sertoli cell. The above changes appeared to be dosage and duration dependent; (2)In the seminiferous tubules of animals receiving CE (1000-2000 ?mol/L testis, 6-14 hr) plus fixation with 10% hypertonic dextrose solutions, usually the germ cells shrinkage and exaggeration of intercellular spaces withint he basal as well asthe adluminal compartments were observed. The tight junction between Sertoli cells were also appeared to be separated by hypertonic action of dextros; (3) The results of tracing experiments showed that lanthanums as tracer could be seen to pass through the Sertoli-Sertoli cell junction of the barrier and enter into the adluminal compartment, the tracer that surrounding the spermatocytes and round spermatids were discernible readily. The above results suggest that cytochalsin E disturbed actin filaments of Sertoli ectoplasmic specialization thus altered the functional integrity of the Sertoli cell barrier. The relationship between the actin filaments and Sertoli cell barrier was discussed.

OBJECTIVE: To explore the mechanism of Chaiqin Chengqi Decoction (CQCQD) in treatment of rats with acute necrotizing pancreatitis (ANP). METHODS: Thirty SD rats were randomly divided into 3 groups: sham-operated (SO) group, ANP group and CQCQD-treated group. ANP was induced by retro-pumping 3.5% sodium cholate to common bile duct. Blood sample was collected from abdominal vein for examination and the pancreatic tissue samples were taken for making pathology section 6 hours later. The pancreatic tissue (HE staining) was observed by light microscope. The content of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) was detected with the method of enzyme-linked immunosorbent assay, and the activation of nuclear factor-kappaB (NF-kappaB) in pancreas was detected by immunohistochemical method. RESULTS: Compared with the SO group, there was dramatic increase in the white blood cell (WBC) counts and AMY level in the ANP group (P<0.05, P<0.01). Compared with the ANP group, the WBC counts and AMY level in CQCQD-treated group were significantly reduced (P<0.05). The edema, inflammatory infiltration, haemorrhage and necrosis scores and total pathological score in the ANP group were obviously higher than those in the SO group (P<0.01). The edema, haemorrhage and inflammatory infiltration scores and the total pathological score in CQCQD-treated group were decreased (P<0.05). The integral optical density of NF-kappaB p65 positive cells of pancreas in CQCQD-treated group was lower than that in the ANP group (P<0.05). CONCLUSION: CQCQD can reduce the content of serum TNF-alpha and IL-6, depress the activation of NF-kappaB, and lessen the pancreatic lesions.

OBJECTIVE: To investigate the occurring mechanism and clinical characteristics of severe acute pancreatitis (SAP) associated with hypoalbuminemia in early stage and its influence on prognosis of SAP and the preventive and therapeutic management of this disease. METHODS: One hundred and thirty-eight cases diagnosed as SAP complicated by hypoalbuminemia in early stage were accepted in our hospital from August 1, 2003 to December 31, 2004, and they were divided into 2 groups according to the level of plasma albumin: mild hypoalbuminemia (30 to 35 g/L) group and severe hypoalbuminemia (<30 g/L) group. The complications in the early stage, related parameters, and the incidence rate of infection and mortality in the later stage were evaluated respectively. RESULTS: The incidence rates of renal dysfunction, shock, cardiovascular failure and gastrointestinal hemorrhage, the score of acute physiology and chronic health evaluation II (APACHE II ) and the frequencies of pulse and breath in the severe hypoalbuminemia group were all higher than those in the mild hypoalbuminemia group (P<0.05 or P<0.01). The differences of incidence rate of hepatic failure and the scores of Ranson and Balthazar CT between these two groups had no statistical significance (P>0.05). The incidence rate of infection and the mortality in the severe hypoalbuminemia group were higher than those in the mild hypoalbuminemia group (P<0.01) in the later stage of SAP. CONCLUSION: Hypoalbuminemia in the early stage can accelerate the deterioration in pathophysiology of SAP. The lower level of the plasma albumin is in the early stage, the more complications and the higher incidence rate of infection and mortality will be in the later stage. To relieve the extent of systemic inflammatory response syndrome (SIRS) and abundant supplement of albumin, amino acid and lipid in time may be crucial to prevent the occurrence and deterioration of hypoalbuminemia.

Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

OBJECTIVETo explore the neuropsychological characteristics of children with attention deficit hyperactivity disorder (ADHD).

METHODSNeuropsychological tests, including visual working memory, Stroop test, digits inverse reciting, vocabulary fluency, Wisconsin card sort test (WCST), and Temporal discounting were used to evaluate the capacity of response inhibition, phonological working memory, visual working memory executive function and delayed satisfying capacity of subjects.

RESULTS1. The ADHD children spent longer time [ADHD-I (84(20), ADHD-C: 98 (31), normal: 70 (28)] to accomplish color naming and made more errors [ADHD-I: 3 (3), ADHD-C: 6 (19), normal: 2 (5)] than the normal control when the color was inconsistent with the word meaning in Stroop test (P < 0.01). 2. The scores of digits reciting [ADHD-I: 3 (3), ADHD-C: 3 (4), normal 4 (4)] inverse was lower in ADHD than in normal control (P < 0.01). 3. The representation of ADHD was poorer than normal control in visual working memory [ADHD-I: 21 (3), ADHD-C: 20 (5), Normal: 20 (3)], and in delayed visual memory [ADHD-I: 19 (5), ADHD-C: 19 (5), Normal: 20 (5)] (P < 0.01). 4. The scores of vocabulary fluency [ADHD-I: 1 (1), ADHD-C: 2 (1), normal: 0 (0)] was lower in ADHD than in normal control (P < 0.01). 5. In WCST, the ADHD children made more errors [ADHD-I :15 (17), ADHD-C: 15 (15), normal: 13 (13)] and less classification [ADHD-I: 5 (4), ADHD-C: 5 (4), normal: 5 (3)] than normal control (P < 0.01). 6. In Temporal discounting, the ADHD children showed significantly more impairments than normal control did (P < 0.01). 7. There was significant difference between the two subtype groups on some tests (P < 0.01).

CONCLUSIONSObvious cognitive impairments were found in children with ADHD, involving poor response inhibition, impaired working memory, dysfunction of planning and set-shifting, and there was no significant difference between the two subtype groups.

Attention Deficit Disorder with Hyperactivity ; classification ; immunology ; physiopathology ; psychology ; Child ; Cognition Disorders ; physiopathology ; psychology ; Humans ; Memory ; Memory Disorders ; immunology ; psychology ; Memory, Short-Term ; physiology ; Neuropsychological Tests

OBJECTIVETo evaluate the value of three-dimensional surface reconstruction of spiral CT-shaded surface display (SSD) to examine the impacted teeth before orthodontic treatment.

METHODSThree-dimensional surface reconstruction of spiral CT and shaded surface display (SSD) was applied to twenty patients whose impacted teeth couldn't be judged clearly through the panorama and occlusal films.

RESULTSThree-dimensional surface reconstruction of spiral CT and SSD could clearly demonstrate the dental surface image including crown, root neck and root bifurcation in three-dimensional way, labial or palatal location, eruption orientation and relation with dentition.

CONCLUSIONIt is concluded that the three-dimensional surface reconstruction is an accurate and effective method to examine impacted teeth before orthodontic treatment.

Female ; Humans ; Male ; Orthodontics ; Tomography, Spiral Computed ; Tomography, X-Ray Computed ; Tooth Root ; Tooth, Impacted

OBJECTIVETo distinguish lingual thyroglossal duct cyst (LTDC) from laryngomalacia in newborn infants.

METHODSData of 10 newborn infants with laryngeal stridor and dyspnea, admitted to the department of neonatology in our hospital during December, 2004 to August, 2007, who were finally diagnosed with LTDC though previously diagnosed as congenital laryngeal stridor in other hospitals, were summarized and analyzed.

RESULTSInspiratory stridor with chest wall retractions was cardinal symptom of newborn with LTDC. A slightly gray and round cyst with smooth surface at the base of the tongue was found with laryngoscopy. On computed tomography examination of larynx, a well-circumscribed lesion with low intensity was detected at the base of the tongue protruding into the air passage. Pathological examination demonstrated that the cyst wall was composed of tabular and columnar epithelium.

CONCLUSIONSLTDC is a common disease in newborns, which is similar to laryngomalacia. For neonates suspected of LTDC, laryngoscopic examination should be performed first, while laryngeal CT scan is an important diagnostic basis. Cyst puncture can ameliorate the symptoms of the patients, while surgical removal is the method of radical cure.

Early Diagnosis ; Female ; Humans ; Infant, Newborn ; Male ; Respiratory Sounds ; Retrospective Studies ; Thyroglossal Cyst ; diagnosis

A new method for establishing ES cell lines from 129/ter. C57BL/6J mice was set up which was characterized by the murine embryonic fibroblast cell(MEF) feeder, the medium of rat heart cell-conditioned medium(RH-CM) for ES cells, and the consecutive digestion by the digestion liquid containing 1% serum. Every group of improved experiments was done with a control of routine method. The results showed that, compared with routine method, the improved way increased the ratio of ES cell lines of 129/ter mice from 11.8% to 33.3%, and of C57BL/6J from 3.7% to 13.3%. The difference is distinct. The passage culture of ES cells showed that, compared with medium added LIF, RH-CM not only inhibited the differentiation of murine ES cells, maintained its dipoild karyotype, but also promote its adherence growth. This kind of culture condition not only maintained the ES cells in an undifferentiated state and their normal dipoild karyotype, but also a series of other characteristics of totipotent embryonic stem cells during extended culture period.
Alkaline Phosphatase ; metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Cell Differentiation ; Cell Division ; Cell Line ; Embryo, Mammalian ; cytology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Rats ; Stem Cells ; physiology

To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.
Adenoviridae ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; Genetic Therapy ; Genetic Vectors ; Humans ; Neoplasms ; metabolism ; pathology ; virology ; Oncolytic Viruses ; genetics ; metabolism ; physiology ; Quality Control ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Virus Replication

Objective:To prepare,purify the recombinant proteins of HSP70-like protein 1 (HSPTOL1) with a large fragment of chicken ovalbumin (OVA),and to investigate the bio-function of the fusion protein,providing a basis for fur- ther study of the effect and the mechanism of HSPTOL1 as an adjuvant.Methods:The vector containing HSP70L1 cDNA and large fragment of OVA was constructed.The expression of OVA-HSP70L1 fusion protein was induced and the products were purified from inclusion bodies by His-Trap metal chelation chromatography and DEAE ion-exchange chromatography. The bio-activity of the fusion protein was examined by detecting its ability to activate dendritic ceils and to promote the se- cretion of cytokines.Results:The vector was successfully constructed and the molecular weight of the fused OVA- HSPTOL1 protein (with a purity of over 95%) was 68 000.The fusion protein effectively promoted the maturation of den- dritic cells and the production of cytokines such as interleukin-12 and tumor necrosis factor-?.Conclusion:HSPTOL1 may be an effective adjuvant in the fusion protein and it may also promote antigen specific Thl type i mmol/Luno-respon- ses.