OBJECTIVETo investigate the effects and mechanisms of diethylhexylphthalate (DEHP) on morphology and function of progenitor Leydig cells (PLC) in rats.

METHODSTwenty pregnant SD rats were randomly divided into 4 groups ( n = 5): normal control group, DEHP low dose group , middle dose group, and high dose group, which were treated from postnatal day (PND) 1 to PND 21 of the pubs with DEHP at the doses of 0, 10, 100, 750 mg/(kg · d) in 0.5 ml of corn oil by gavage respectively. At the end of the treatment, the male pups were killed and blood samples were collected for determination of serum testosterone concentration by chemiluminescence method. The body weight, testis weight and anogenital distance (AGD) were measured. The morphology of PLC was observed by light and transmission electron microscopy. The protein expression of steroidogenic acute regulatory protein(StAR) in PLC was determined by immunohistochemistry. The mRNA expression of insulin-like growth factor-I (IGF-I) in the testis was assayed by real-time PCR.

RESULTSCompared with normal control group, the serum testosterone and AGD of male pubs from the middle and high dose groups were declined significantly (P < 0.01), the testis weight and body weight from high dose group were decreased significantly (P < 0.01), while the testis weight increased in the low dose group (P < 0.05). Under light microscope, PLC showed hyperplasia and cluster aggregation in the low dose group and focal hyperplasia in the middle and high dose group. The spermatogenic cells in seminiferous tubules showed decrease, apoptosis and unfix in the high dose group. Under transmission electron microscope, the PLC showed decreased lipid droplets, smooth endoplasmic reticulum and mitochondriae in the treated group. The mRNA expression of IGF-I increased in the low dose group, and the protein expression of StAR decreased in the middle and high dose group.

CONCLUSIONLactating exposure to DEHP may interfere with the synthesis of testosterone of PLC in male pubs, the decrease of StAR and the damage of PLC may be involved in it.

Animals ; Body Weight ; Diethylhexyl Phthalate ; adverse effects ; Female ; Germ Cells ; drug effects ; Insulin-Like Growth Factor I ; metabolism ; Lactation ; Leydig Cells ; cytology ; drug effects ; Male ; Organ Size ; Phosphoproteins ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects ; Testis ; Testosterone ; blood

Objective To study the COLIXA3 gene polymorphism of patients with fluorosis and to explore the pathogenesis of COLIXA3 gene in endemic fluorosis.Methods Fifty one cases of patients with drinking-water borne fluorosis were selected as the case group in Xinzhou city,Shanxi province and 28 cases of healthy people were as the control group.Dental fluorosis was detected by Dean method and skeletal fluorosis was examined by X-ray.COLIXA3 of exon 5 gene product of 103 points was amplified by PCR and the gene locus genotype was sequenced.Results Ten cases of mild dental fluorosis,14 cases of moderate dental fluorosis,15 cases of severe dental fluorosis were detected among the 51 patients.The control group was free of dental fluorosis.All the 51 cases of patients with fluorosis had varying degrees of skeletal fluorosis,mainly osteosclerosis lesions,accounting for 86.27％(44/51 ),and mild skeletal fluorosis patients were all osteosclerosis lesions,and osteosclerosis lesions and multiple skeletal lesions were found among moderate and severe skeletal fluorosis patients in the case group,while control group had no skeletal fluorosis.The differences between genotypes of frequency distribution of AA,Aa,aa of COLIXA3 of case and control groups were not statistically significant [96.08％(49/51 ),3.92％(2/51 ),0.00％(0/51) and 96.43％(27/28),3.57％(1/28),0.00％(0/28),x2 =0.94,P ＞ 0.05].ConclusionsCOLIXA3 gene polymorphism is not significantly correlated to fluorosis.

OBJECTIVETo explore the role of paternal veto cells in preventing graft-versus-host disease (GVHD) after related HLA- haploidentical stem cell transplantation in mice.

METHODSMHC-haploidentical recipient B6CF1(H-2 b/d) mice pretreated with total body irradiation at 9.0 Gy for 4 h before transplantation. The recipient mice were divided into 4 groups, and in the irradiation group, only injection of 0.3 ml D-Hank's liquid was given through the tail vein; in the control group, the mice received injection through the tail vein of 4.5x10(6) bone marrow cells mixed with 3.0x10(7) spleen cells from C57BL/6 mice without the preventive measures of GVHD; the mice in the two experiment groups received cell transplantation in the same manner, and on day 4 after transplantation, 5.0x10(6) and 1.0x10(7)spleen cells from BALB/c mice were injected through tail vein, respectively. The hematopoietic recovery, engraftment and GVHD of the recipient mice were observed.

RESULTSWithout any treatment, all mice in the control group developed GVHD and died after transplantation. In the 10 mice with injection of 5.0x10(6) spleen cells, GVHD occurred in 5 mice with a 30-day survival rate of 50%; the median survival time of the mice with GVHD was 20 days, significantly longer than that of the control mice (14 days, P<0.05). In the 10 mice injected with 1.0x10(7) spleen cells, 2 developed GVHD and the 30-day survival rate was 80% (8/10) with a median survival time of 30 days, significantly longer than that of mice with injection of 5.0x10(6) spleen cells and the control mice (P<0.05).

CONCLUSIONPaternal veto cell transplantation can decrease the occurrence of GVHD after related HLA haploidentical stem cell transplantation in mice.

Animals ; Cell Transplantation ; methods ; Female ; Graft vs Host Disease ; etiology ; prevention & control ; HLA Antigens ; immunology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Spleen ; cytology ; immunology ; T-Lymphocytes ; cytology ; drug effects ; Transplantation, Homologous

OBJECTIVETo compare the efficacy of nasal synchronized intermittent mandatory ventilation (nSIMV) and nasal continuous positive airway pressure (nCPAP) in preterm infants with respiratory distress syndrome (RDS).

METHODSFifty preterm infants with RDS who received pulmonary surfactant were randomized to nSIMV and nCPAP groups after extubation. Clinical signs, symptoms and blood gas results following nSIMV or nCPAP were compared in the two groups.

RESULTSCompared with the nCPAP group, the nSIMV group had a lower incidence of failure respiratory support (24% vs 60%; P<0.05), a lower incidence of hypercarbonia (12% vs 40%; P<0.05) and a lower incidence of hypoxia (24% vs 36%; P<0.05).

CONCLUSIONSnSIMV is more effective in respiratory support in preterm infants with RDS.

Continuous Positive Airway Pressure ; methods ; Humans ; Infant, Newborn ; Infant, Premature ; Intermittent Positive-Pressure Ventilation ; methods ; Respiratory Distress Syndrome, Newborn ; therapy

OBJECTIVETo explore the role of TJU103 in preventing graft-versus-host disease (GVHD) after allogeneic stem cell transplantation in mice.

METHODSBALB/c mouse splenic lymphocytes were collected and treated by mitomycin as the activating cells and the C57BL/6 mouse splenic lymphocytes as the reacting cells. In the experimental groups, the effect of TJU103 on the proliferative response of T cells was observed. BALB/c(H-2d) and CB6F1(H-2d/b) mice were used as the MHC-full-mismatched recipients and MHC-haplo-identical recipients, respectively, and pretreated by total body irradiation at 9.0 Gy before transplantation. For the recipients of the irradiation group, 0.3 ml D-Hank's solution was injected through the tail vein without cell transplantation, the recipients of the control group received injection of 4.5x10(6) bone marrow cells mixed with 3.0x10(7) spleen cells from C57BL/6 mice through the tail vein, and those in the experimental group received cell transplantation in the same manner with also injection via the tail vein of 25 microg/ml TJU103, which was subsequently injected intraperitoneally for 7 consecutive days at daily dose of 50 microg. The hematopoietic recovery, engraftment and GVHD of the recipients were observed.

RESULTSTJU103 resulted in a dose-dependent inhibition of T cell proliferation in mixed lymphocyte reaction (MLR), and nearly 83% inhibition of the proliferative response was observed with the addition of 25 microg/ml of TJU103. Without any treatment, the occurrence of GVHD and death rate in the control group was both 10/10. Daily injection of TJU103 at 50 microg for the initial post-transplantation week protected the mice from GVHD. In the MHC-full-mismatched model, the incidence of GVHD and survival rate on day 30 of the experiment group was 2/10 and 8/10, showing significant difference from those in the control group (P<0.01). The median survival time (MST) was 30 days in the experimental group versus 15 days in the control group (P<0.05). In the MHC-haplo-identical model, the incidence of GVHD and the survival rate on day 30 of the experimental group was 1/10 and 9/10, which were significantly different from the control group (P<0.01). The MST was 30 days in the experimental group versus 14 days in the control group (P<0.05).

CONCLUSIONTJU103 is capable of markedly inhibiting T cell proliferative response in vitro and can decrease GVHD incidence after allogeneic stem cell transplantation in mice.

Animals ; Cell Proliferation ; drug effects ; Female ; Graft vs Host Disease ; etiology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Organic Chemicals ; pharmacology ; therapeutic use ; Stem Cell Transplantation ; adverse effects ; methods ; T-Lymphocytes ; cytology ; drug effects ; Transplantation, Homologous

OBJECTIVETo analyze the related factors of neck recurrence and regularity of cervical lymph nodes metastasis of pathologically node positive (pN+) tongue squamous cell carcinoma(SCC) and explore the neck treatment strategy for pN+ tongue SCC.

METHODSClinical and follow-up data of 138 patients with pN+ oral tongue SCC from Jan. 1991 to Dec. 2008 were reviewed. Distribution of neck metastatic and recurrent lymph nodes were analyzed. The influencing factors of neck recurrence of pN+ tongue SCC were analyzed.

RESULTSAll patients were followed over two years or until death. Using Kaplan-Meier method, the 3-year and 5-year overall survival rates were 46.4% and 36.2% respectively. Two hundred and three levels of 138 patients had metastasis and the involvement frequency of ipsilateral I, II, III reached to 94.6%. Sixty-six levels of 47 patients had neck recurrences and the involvement frequency of ipsilateral I, II, III reached to 77.3%. pT stage, pN stage, pTNM stage, extracapsular spread (ECS) of cervical lymph nodes were relevant to the neck recurrence of pN+ tongue SCC (all P < 0.05). When ECS of cervical lymph nodes was present, the neck recurrence rate of patients with postoperative radiation was lower than patients without postoperative radiation, but P value failed to demonstrate significant difference (P = 0.076). There were no significant difference of neck recurrence rates between different neck dissection methods (P > 0.05). Multivariate Cox analysis showed that pTNM stage and ECS of cervical lymph nodes were the independent prognostic factors of pN+ oral tongue SCC.

CONCLUSIONSpT stage, pN stage, pTNM stage, ECS of cervical lymph nodes were the influencing factors of neck recurrence of pN+ tongue SCC. Postoperative radiation may reduce the neck recurrence rate when ECS was present. There was no difference of the neck recurrence rate between modified neck dissection (MRND) and radical neck dissection (RND) and when the non-lymphatic structures were not involved, MRND should attempted. Metastatic and recurrent lymph nodes of pN+ tongue SCC were mostly distributed in ipsilateral I, II, III level and selective neck dissection (SND) can be applied to pN+ tongue SCC.

Carcinoma, Squamous Cell ; Humans ; Lymphatic Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Retrospective Studies

OBJECTIVETo summarize the clinical changing characters of the clinical markers after interferon treatment in chronic hepatitis B (CHB) and make out practical indexes to predict the effect.

METHODS150 CHB patients were randomly divided into two groups: therapeutic group (90) and control group (60) in the prospective controlled trial. The levels of endogenous interferon before treatment, interferon antibody at the end of the second month and fourth month after treatment, alanine aminotransferase (ALT) and HBV DNA in the serum were detected. Then the data was analysed to find out indexes for predicting the effect.

RESULTS(1) The clearance rate of HBeAg had no significant difference in age except for 20 - 30 and 30 - 40 (t > 2.331 2, P < 0.01). (2) It was more effective if ALT level was higher than 400 U/L before treatment and it decreased more than 50% two months after treatment. (3) The patients whose HBV DNA was negative (dot hybridization) or less than 10(6) copies/ml before treatment had higher rate of HBeAg clearance. (4) There was no effect on patients whose interferon antibody turned positive at the end of the second month. (5)A predictive method of comprehensive factors was made out, whose sensitivity, specificity, and accuracy were 80%, 100% and 90%, respectively.

CONCLUSIONThe clinical characters of these Chinese patients are different from those of the westerners and the effects of interferon have close relation to the levels of ALT, HBV DNA and interferon antibody.

Adjuvants, Immunologic ; administration & dosage ; therapeutic use ; Adolescent ; Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; drug therapy ; physiopathology ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Prospective Studies

BACKGROUNDTo study the molecular characteristics of YN92-4 strain isolated from mosquitoes in Yunnan Province and define its classification.

METHODSThe S segment of YN92-4 strain was amplified and sequenced by 2 different sets of primers. The phylogenic tree of S fragment was constructed by Phylip bio-software. The amino acid sequences of N and NSs proteins were also studied.

RESULTSYN92-4 strain could be amplified by 2 sets of primers respectively, S segment showed a highest homology with Batai virus (X73464), reached 96.4%, the homology of protein N and NSs amio-acid sequence with Batai virus was 99.1% and 98% respectively.

CONCLUSIONThe YN92-4 strain belongs to Batai virus, this is the first report of molecular biological identification of Batai virus in China.

Amino Acid Sequence ; Animals ; Bunyamwera virus ; classification ; genetics ; isolation & purification ; China ; Culicidae ; virology ; DNA, Complementary ; chemistry ; genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

BACKGROUNDIschaemic reperfusion injury (IRI) is inevitable during major liver surgery. Ischaemic preconditioning (IPC) has been proven an effective intervention against hepatic IRI. Recently, it was demonstrated that ischaemic postconditioning (IPO) provided effective cardioprotection on IRI. We evaluated the protective effects of IPO on warm/cold IRI in rat liver by a comparison with IPC and assessed the role of apoptosis in the process.

METHODSWarm IRI model (clamping hepatic pedicle for 30 minutes) and cold IRI model (orthotopic liver transplantation with 2 hours cold storage) were established. Each model consisted of 3 groups: (1) control group, normal warm/cold IRI; (2) IPC group, 5 minutes of ischaemia followed by 5 minutes of reperfusion twice prior to warm/cold IRI; (3) IPO group, 30 seconds of reperfusion followed by 30 seconds of reocclusion for three times after warm/cold ischaemia. The levels of serum transaminase, glucose, and gamma glutamyltransferase (GGT) in bile, histopathological examination, apoptotic activity of hepatocyte, and apoptosis related protein Fas, at 3 hours after operation were compared. Survival rates one week after intervention were also compared.

RESULTSIPO and IPC protected the functions of hepatocytes and biliary epithelial cells, inhibited the hepatocellular apoptosis by preventing expression of Fas gene, and elevated the one week survival rate compared with control group in both models (P < 0.05). IPO and IPC groups were comparable in levels of serum transaminase levels, glucose, and GGT in bile, Fas positive expression index, and one week survival. In cold ischaemic models, IPO had lower apoptotic index than IPC (P < 0.05).

CONCLUSIONCompared with ischaemic preconditioning, ischaemic postconditioning is associated with comparable protections of rat liver from warm or cold ischaemic reperfusion injury.

Animals ; Apoptosis ; Ischemic Preconditioning ; Liver ; blood supply ; pathology ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Survival Rate ; fas Receptor ; genetics

OBJECTIVETo evaluate the feasibility of HLA haploidentical peripheral blood hematopoietic stem cell transplantation (PBSCT) for patients with β thalassemia major.

METHODSSixteen patients with β thalassemia major received HLA haploidentical PBSCT from parents. Two conditioning regimens were used. Regimen A was adopted before December 2007, which consisted of fludarabine (total 150 mg/m²), busulfex (total 520 mg/m²), cyclophosphamide (CTX, total 100 mg/kg), antithymocyte globulin (ATG, total 10 mg/kg) and total body irradiation of 3 Gy. Regimen B was adopted after December 2007, which consisted of fludarabine (total 240 mg/m²), busulfex (total 520 mg/m²), CTX (total 100 mg/kg), and ATG (total 10 mg/kg). Combination of cyclosporin (CsA), methotrexate (MTX) and mycophenolate mofetil (MMF) were used for prophylaxis of graft-versus-host disease (GVHD).

RESULTSOf 16 patients, 14 (87.5%) had sustained engraftment. The median days of neutrophil exceeding 0.5 × 10⁹/L and platelet exceeding 20 × 10⁹/L were 13 days (range 10 - 17 days) and 15 days (range 14 - 20 days) after PBSCT, respectively. Complete chimerism was achieved in all the 14 patients at one month after PBSCT. One patient lost his graft with autologous reconstitution 52 days after transplantation. Four patients had grade II-IV acute GVHD and one patient had chronic extensive GVHD. In the 49-month median follow-up duration, 13 of 16 patients were alive in disease-free situation.

CONCLUSIONHLA haploidentical PBSCT, which could provide stable and sustained engraftment for thalassemia major patients with no HLA identical donor, is a promising treatment strategy.

Child ; Child, Preschool ; Female ; HLA Antigens ; genetics ; Haploidy ; Humans ; Male ; Peripheral Blood Stem Cell Transplantation ; Tissue Donors ; beta-Thalassemia ; therapy