Using the chromosomal DNA of an ice nucleation active bacterium Erwinia ananas 110 as template, an ice nucleation active (ina) gene was amplified by PCR with Taq plusI DNA polymerase. After sequencing and compared with reported ina genes, the cloned gene was identified as a new ina gene and was registered in GenBank at the accession number of AF387802. The new ina gene, named as iceA, has 3921 bp for its coding region, which encodes 1306 amino acids consisting of repetitive segment (R-domain, 1104aa), which is flanked by N-and C-terminal sequences, with 161 aa and 41aa, respectively.

Objective To analyze the clinical characteristics of infants with functional constipation(FC)relating to food allergy(FA)and explore its ways of diagnosis and therapy.Methods The detail clinical information of the patients who corresponded to the criteria of diagnosis of FC and FA simultaneous from Mar.2007 to Dec.2009 were reviewed and analyzed.There were 496 cases of FA,in the FA joint clinic,gastroenterology clinic and pediatric ward,56 cases of them had the symptoms of FC(56/496 cases,11%),diets free and amino acid-based formulae were the treatments of the patients.The 56 cases were analyzed with skin-prick test(SPT),serum special IgE and serum special IgG,and the risk factors attributed to the no response to treatments with Logistic regression.Results Fifty-six cases opened the bowel less than twice a week,and companied with pain,8 patients with soiling once a week at least,10 patients with fecal retention,4 patients with large fecal in rectum.Mother or father with the history of allergy were 22 cases,17 cases,separately,both of mother and father with the history of allergy were 17 cases.The results of SPT,serum special IgE and serum special IgG showed that the patients with different allergens,cow's milk,egg,cod,especially.The patients with FC relating to FA were breast-feed,battle-feed or mix-feed.Treatment relied on amino acid-based formulae,as well as elimination diets in infants,80%of them(45/56 cases)improved.Introducing solid foods,the age of upsetting constipation and premature were the risk factors contributed to the no response to the diet free treatment.Conclusion FA should be consi-dered to contribute to the infants presenting with FC.Diets free and amino acid-based formulae may be effective.

OBJECTIVE: To explore the distribution of inflammatory cells and positive expression of P-se- lectin glycoprotein ligand-1 (PSGL-1) in infant brainstem tissue from hand-foot-mouth disease related fatal brainstem encephalitis. METHODS: Twenty brainstem samples from infants suffered from brainstem en- cephalitis were collected as the experimental group. Ten brainstem samples from infants died of non- brain diseases and injuries were collected as the control group. The distribution of inflammatory cells and the expression of PSGL-1 in the two groups were examined by immunohistochemical method. The characteristics of the positive cells were observed. RESULTS: In brainstem tissue of the experimental group, there were sleeve infiltrations of inflammatory cells around the vessels and in the glial nodule. Microglia was the most and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference among microglias, neutrophils and lymphocytes (P < 0.05). There was no sleeve infiltration in the control group. PSGL-1 protein was expressed widely in inflammatory cells in the experimental group, especially in the inflammatory cells around the vessels and in the glial nodule. But PSGL-1 positive staining could be observed significantly less in the control group comparing with the experimental group (P < 0.05). CONCLUSION Microglia is the main type of inflammatory cells involved in the progress of the fatal disease. Moreover, PSGL-1 could participate in the pathogenesis of hand-foot-mouth disease related fatal brainstem encephalitis.
Brain Stem/pathology* ; Encephalitis/pathology* ; Hand, Foot and Mouth Disease/pathology* ; Humans ; Infant ; Membrane Glycoproteins/metabolism* ; Microglia/pathology* ; Neutrophils/pathology*

Objective To investigate the risk factors of low birth weight infants with food allergy.Methods Forty-nine food allergy children who had been admitted to the neonatal intensive care unit(NICU) of Peking University Third Hospital from Jan.1,2007 to Dec.31,2011 and followed up in the Pediatrics Allergy Clinic after discharged from hospital were involved as food allergy group.Forty-nine children who had been admitted to the NICU of Peking University Third Hospital at the same time and followed up in the Child Health Care Clinic after discharged from hospital but had no manifestations of food allergy were selected by stratified sampling as control group.A questionnaire was designed to record the children's gender,gestation age,mode of delivery,feeding patterns after discharging from hospital,tobacco and alcohol consumption of their parents,allergy history of their parents,the intake of egg of the mother during lactation and the children's introduction of egg.At the same time,their hospital notes were investigated.The diseases they had and the application of parental nutrition,pulmonary surfactant and ventilator were all analyzed.Quantitative data were analyzed with Chi-square,qualitative data were analyzed with t-test.The risk factors of food allergy of the low birth weight infants were examined by unconditional Logistic regression.Results The birth weight,gender,mode of delivery,feeding patterns after discharging from hospital,parents,exposure to tobacco smoke and alcohol,the intake of egg of the mother during lactation and the children's introduction of egg between the 2 groups were no significantly different.The diseases they got and the application of parental nutrition,pulmonary surfactant and ventilator when they in hospital were no significantly different,too.The gestation age,parental history of allergy between the 2 groups were significantly different.The proportion of children of 33-35 weeks and 36-37 weeks gestation age in food allergy group were significantly higher than those in control group,while the 29-32 weeks gestation age were significantly lower than those in control group (x2 =10.312,P =0.016 ;x2 =20.753,P =0.000).But by unconditional Logistic regression analysis,only the one or both parents,history of allergy was the risk factor for the development of food allergy of the low birth weight infants(OR =5.574,P =0.004;OR =14.487,P =0.000).Conclusion Parents history of allergy is related to the development of food allergy of the low birth weight infants.

OBJECTIVETo investigate the expression and significance of CD151 in pituitary adenomas.

METHODSThirty-six pituitary adenomas were collected immediately after surgery together with five normal pituitary tissue. Real time-PCR, Western blot and immunohistochemistry analysis were performed to detect the expression of CD151 mRNA and protein in thirty-six pituitary adenomases and five normal pituitary tissues.

RESULTSThe expression of CD151 in all pituitary adenomases was observed to be significantly higher than that in normal pituitary tissues by Western blot, real time PCR, and immunohistochemistry analysis (P < 0.01). The expression levels of protein and mRNA in invasive pituitary adenomas were much higher than those in non-invasive pituitary adenomas (P < 0.01).

CONCLUSIONThe results suggested that the expression of CD151 was closely correlated with malignant degree of pituitary adenomas, which indicated the expression of CD151 was intimately correlated with occurrence and development of pituitary adenomas. Detecting CD151 might be a vital index to predict prognosis of pituitary adenomas.

Adenoma ; metabolism ; Blotting, Western ; Humans ; Immunohistochemistry ; Pituitary Gland ; pathology ; Pituitary Neoplasms ; metabolism ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Tetraspanin 24 ; metabolism

Fanconi anaemia (FA) is an autosomal recessive inherited disorder caused by defects in hematopoietic stem cells. The clinical manifestations of FA are diverse and complicated. FA cells display high hypersensitivity to agents which produce interstrand DNA cross-links such as mitomycin C (MMC) or diepoxybutane (DEB). At least eight complementation groups with defects in eight genes (FANCA, FANCB, FANCC, FANCD(1), FANCD(2), FANCE, FANCF and FANCG) have been identified by gene analysis. Six genes (corresponding to subtypes A, C, D(2), E, F and G) have been coloned, and the encoded FA proteins interact in a common cellular pathway - "FA Pathway", through which modulate DNA repair. The progress of research on FA molecular mechanism provides gene therapy of FA with theory basis. FA cells transduced with the use of retrovirus carring the normal FA gene cDNA manifestate phenotypic correction of hypersensitivity to DNA cross-linking agents, such as MMC. In this review the clinical manifestations and gene composition of FA, and the functions of encoded FA proteins were summarized. The hematopoietic stem cell transplantation and gene therapy for FA patients were discussed.
Cell Cycle Proteins ; DNA-Binding Proteins ; Fanconi Anemia ; genetics ; metabolism ; therapy ; Fanconi Anemia Complementation Group C Protein ; Fanconi Anemia Complementation Group D2 Protein ; Fanconi Anemia Complementation Group Proteins ; Genetic Therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; Mutation ; Nuclear Proteins ; genetics ; Proteins ; analysis ; genetics

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.
Carcinoma, Hepatocellular ; immunology ; pathology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Killer Cells, Lymphokine-Activated ; cytology ; immunology ; Liver Neoplasms ; immunology ; pathology

OBJECTIVETo explore the apoptosis and the express of proliferating cell nuclear antigen (PCNA) in spermatogenic cells, and study generation function of the rabbit after scrotal reconstruction with flaps.

METHODSThe 48 New Zealand white rabbits were randomly divided into three groups as experimental group (the scrotum reconstructed with flaps, n = 48), the control group (the sham operated group, n = 24) and the blank group (n = 18). The apoptosis and the expression of PCNA in the spermatogenic cells were detected with TUNEL and the immunohistochemistry from the 3rd to the 8th week after operation. 8 weeks later, 12 animals in each group were fed respectively with one female rabbit to observe the procreation.

RESULTSThe apoptotic index of the spermatogenic cells in blank group was 7.73 +/- 4.95. 3 weeks after operation, the apoptotic index of spermatogenic cells was 22.59 +/- 3.04 in the experimental group, and 21.13 +/- 1.68 in control group, showing no significant difference between the two groups (P > 0.05). 8 weeks after operation, the apoptotic index of spermatogenic cells was 71.85 +/- 2.69 in the experimental group, and 13.64 +/- 2.09 in control group, show a significant difference between them (P < 0.05). The apoptotic index in experimental group increased gradually from the 3rd to 8th week after scrotal reconstruction , which was markedly higher than that in the blank group (P < 0.05). The apoptotic index in control group was higher than that in the blank group at the 3rd week (P < 0.05), but not at the 8th week (P > 0.05). The proliferation index of spermatogenic cells was 9.32 +/- 9.30 and 12.52 +/- 3.87 in experimental group at the 3rd and 4th week, respectively, which was significantly lower than that in blank group (43.07 +/- 2.25) and control group (45.69 +/- 4.98) at the 3rd week (P < 0.05). The proliferation index of spermatogenic cells was 46.98 +/- 18.92 and 49.53 +/- 9.79 in experimental group at the 7th and 8th week, respectively, 39.90 +/- 5.10 in control group at the 8th week, showing no difference between the two groups (P > 0.05). The proliferation index of spermatogenic cells in control group at the 3rd and 8th week was not different from that in the blank group (P > 0.05). The female pairing rabbits in the blank and control group were all pregnant, and the average childbirths were 6.0 +/- 1.28 and 5.92 +/- 1.31 respectively, with no difference between the two groups (P > 0.05). All the female pairing rabbits in the experimental group were not pregnant, showing a significant difference from those in the blank and control group (P < 0.05).

CONCLUSIONSThe rabbit generation functional disturbance after scrotal reconstruction with flaps is due to the excessive apoptosis of spermatogenic cell. The spermatogenic cell proliferation is affected only in the early postoperative period, but can recover later.

Animals ; Apoptosis ; Cell Proliferation ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits ; Scrotum ; surgery ; Seminiferous Epithelium ; cytology ; Skin Transplantation ; Surgical Flaps

Objective To examine short-term outcomes and long-term survival of elderly patients (aged over 80 years) with colorectal cancer who received laparoscopic versus open surgery. Methods A total of 313 patients over 80 years old with colorectal cancer who underwent radical surgery were included.According to the surgical method, all patients were divided into open-surgery group (n=143) and laparoscopic surgery group (n=170).Baseline data were balanced between the two groups by using propensity score matching.Kaplan-Meier was used to draw the survival curve, and survival was compared by Log rank tests.Cox proportional risk model was used to analyze the effects of all factors on overall survival (OS) and disease-free survival (DFS). Results After matching, 93 patients were included in each group.The mean intraoperative blood loss, the incidence of overall postoperative complications and gradeⅠ-Ⅱ complications in the laparoscopic surgery group were significantly lower than those in the open surgery group (all P < 0.05).The time to first flatus, the time to oral feeding, and postoperative hospital stays in the laparoscopic surgery group were significantly shorter than those in the open surgery group (all P < 0.05).The mean number of lymph-node dissection was also significantly higher in the laparoscopic surgery group than in the open surgery group (P=0.030).Patients in both groups had similar 5-year OS (P=0.594) and DFS (P=0.295).Multivariate Cox prognostic analysis showed that CEA level > 5 ng/ml, pathological TNM stage Ⅲ, and perineural invasion were independent risk factors for poor OS and DFS. Conclusion Compared with open surgery, laparoscopic surgery can provide better short-term advantages and similar long-term outcomes for colorectal cancer patients over 80 years of age.

OBJECTIVETo investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects at forepart of feet.

METHODSFrom January 2004 to October 2008,15 cases of skin defects at forepart of feet were repaired with reverse saphenous nerve neurocutaneous flaps. The flap size ranged from 3.5 cm x 3.0 cm to 8 cm x 5 cm. The wounds at donor site were closed with skin graft.

RESULTSAll the flaps survived completely with no ulcer at the donor site. 10 patients were followed up for 1 to approximately 9 months. The skin color and texture were satisfactory. The patients could walk very well.

CONCLUSIONSIt is reliable to repair the skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps. It is easily performed with less morbidity.

Adolescent ; Adult ; Child ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Skin ; injuries ; Skin Transplantation ; Surgical Flaps ; blood supply ; innervation ; Young Adult