OBJECTIVETo investigate the functional relations between the putative proteins YpCD1.08, YpCD1.09, YpCD1.16 encoded in pCD1 plasmid of Yersinia pestis and its type III secretion system (T3SS).

METHODSMutants of YpCD1.08, YpCD1.09, YpCD1.16 were constructed using λ-Red recombinant system. The growth curves of the mutant strains cultivated in TMH medium with or without calcium at 26 °C and 37 °C were determined to analyze the low calcium response phenotype. The transcription levels of ΔYpCD1.08, ΔYpCD1.09, ΔYpCD1.16 in Yersinia pestis and the dependence to temperature were determined using real time RT-PCR after cultivation at 26 °C and 37 °C and extraction of RNA. A β-lactamases reporter system was adopted to study the influence of these genes on the translocation of effector YopE of T3SS.

RESULTSWhen grown in TMH medium without calcium at 26 °C and 37 °C, the growth curve of the YpCD1.08, YpCD1.09, YpCD1.16 mutants were similar to that of the wild-type strain, indicating that the low calcium response of all the mutants were normal. The ratios of YpCD1.08, YpCD1.09, YpCD1.16 gene transcriptional level at 37 °C and 26 °C were 2.3 ± 0.3, 2.3 ± 0.5 and 3.2 ± 0.7, respectively, indicating that these genes were transcribed in Yersinia pestis and their transcription regulations showed a temperature-dependence that was consistent with the well established temperature-dependent expression of Yersinia T3SS genes. The β-lactamases reporter assays demonstrated that ΔYpCD1.08 could translocate much higher level of YopE into HeLa cells, since that the light intensity ratio of 477/520 nm at 140 min was 2.5, whereas it was 1.8 for the wild-type strain, and the values in ΔYpCD1.09 and ΔYpCD1.16 were similar to the wild-type strain.

CONCLUSIONYpCD1.08, YpCD1.09, YpCD1.16 gene are likely to be the new members of T3SS, and the putative protein YpCD1.08 could play some roles in YopE secretion and translocation.

Bacterial Outer Membrane Proteins ; secretion ; Bacterial Secretion Systems ; genetics ; Genes, Bacterial ; Plasmids ; Protein Interaction Mapping ; Yersinia pestis ; genetics ; metabolism ; pathogenicity

Objective To investigate the treatment effects of operation and nerve growth factor on adult multisegmental cervical spinal cord injury without fracture dislocation.Methods Sixty-eight patients with multiple segmental cervical spinal cord injury without fracture dislocation,admitted to our hospital from January 2004 to May 2011,were chosen in our study; according to the will of the patients,18 patients received conservative treatment (group A),25 patients were treated by posterior single open-door laminoplasty (group B) and the other 25 patients were treated with posterior single open-door laminoplasty combined with nerve growth factor (group C,once daily for 4 weeks); their clinical data were retrospectively analyzed.Follow up was performed at 3,6 and 12 months after the treatments;Japanese Orthopaedic Association (JOA) scale was employed to evaluate the clinical efficacy.Results Three,6,12 months after the treatments,the JOA scale scores were statistically different among the three groups (P＜0.05); the scores in group B and C were significantly higher than those in group A at all time points (P＜0.05); 6 and 12 months after the treatmemts,the JOA scale scores in group C were significantly higher than those in group B (P＜0.05).Conclusion To patients with multisegmental cervical spinal cord injury without fracture dislocation,conservative treatment can make the spinal cord function partially restored,but the effect is limited; exogenous nerve growth factor has good repairing effect on spinal cord injury.

OBJECTIVETo investigate the function of dendritic cells (DC) of patients with immune related pancytopenia (IRP) and explore the role of DC in IRP.

METHODSThe expression of CD80 and CD86 on myeloid DC (mDC, Lin-HLA-DR(+) CD11c(+) cells) and plasmacytoid DC (pDC, Lin-HLA-DR(+) CD123(+) cells) of 65 IRP (37 untreated and 28 remitted) patients and 17 healthy controls were analyzed by flow cytometry.

RESULTSThe expression of CD86 on pDC was (82.47 ± 13.17)% in untreated group and (60.08 ± 14.29)% in remission group, which were significantly higher than that of controls (47.95 ± 18.59)% (P < 0.05), while the expression in untreated group was higher than that of remission group (P < 0.05). The expression of CD80 on pDC was (6.31 ± 4.49)% in untreated group, which was significantly higher than that of remitted patients (3.09 ± 2.93)% and controls (2.33 ± 2.25)% (P < 0.05). The expression of CD86 on mDC was (97.06 ± 4.82)% in untreated group and (91.35 ± 12.20)% in control group, while the expression in untreated group was higher than that of control group (P < 0.05). The expression of CD80 on mDC was (6.20 ± 5.44)% in untreated group and (3.97 ± 3.24)% in remission group, which were significantly higher than that of controls (1.86 ± 1.73)% (P < 0.05). The expression of CD86 on pDC was negatively correlated to Th1/Th2 (r = -0.733, P < 0.05), it was positively correlated to the antibody on membrane of BMMNC (r = 0.283, P < 0.05) and the quantity of CD5(+)B cells (r = 0.436, P < 0.05), while it was negatively correlated to the level of hemoglobin, platelets and white blood cells (r = -0.539, P < 0.05; r = -0.519, P < 0.05; r = -0.567, P < 0.05, respectively). The expression of CD80 on pDC was negatively correlated to the level of hemoglobin and platelets (r = -0.431, P < 0.05; r = -0.464, P < 0.05).

CONCLUSIONThe function of pDC in PB of IRP were strengthened, which was relevant to the immunopathogenesis of IRP.

Adolescent ; Adult ; Autoimmune Diseases ; complications ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; metabolism ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; etiology ; pathology ; Young Adult

BACKGROUNDThe nocturnal nondipping and elevated morning blood pressure (BP) in patients with obstructive sleep apnea syndrome (OSAS) have not yet been well investigated in Chinese patients. This study aimed to describe the BP profile, and to elucidate the relationships between daytime BP and nighttime BP, and between evening BP and morning BP in patients with OSAS.

METHODSTwenty teaching hospital sleep centers in China were organized by the Chinese Medical Association to participate in this study and 2297 patients were recruited between January 2004 and April 2006. BP assessments were made at four time points (daytime, evening, nighttime and morning) and polysomnography (PSG) was performed and subjects were classified into four groups by their apnea-hypopnea index (AHI): control, n = 213 with AHI < 5; mild, n = 420 with AHI ≥ 5 and < 15; moderate, n = 460 with AHI ≥ 15 and < 30; and severe, n = 1204 with AHI ≥ 30. SPSS 11.5 software package was used for statistical analysis and figure drawing.

RESULTSAll the average daytime, nighttime, evening and morning BPs were positively correlated with AHI and negatively correlated with nadir nocturnal oxygen saturation. The ratios of nighttime/daytime and morning/evening average BP were positively correlated with AHI. The ratio of nighttime/daytime systolic BP became a "reversed BP dipping" pattern until the classification reached severe, while the ratio of nighttime/daytime diastolic BP became reversed at moderate. Similarly, the ratio of morning/evening diastolic BP becomes reversed even at mild.

CONCLUSIONSOSAS may result in higher BP levels at all four time points. The ratios of nighttime/daytime and morning/evening BP increase with increased AHI. The increasing of diastolic BP, which is inclined to rise more quickly, is not parallel with increasing systolic BP.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anthropometry ; Blood Pressure ; physiology ; Female ; Humans ; Hypertension ; physiopathology ; Male ; Middle Aged ; Sleep Apnea, Obstructive ; physiopathology ; Young Adult

This study was aimed to investigate the quantity and subtypes of dendritic cells (DC) in patients with immune related pancytopenia (IRP) and to explore the role of DC in pathogenesis of IRP. The quantity of plasmacytoid dendritic cells (pDC, Lin(-)HLA-DR(+) CD123(+) cells) and myeloid dendritic cells (mDC, Lin(-)HLA-DR(+) CD11c(+)cells) in peripheral blood of 65 patients with IRP (37 new diagnosed and 28 remitted) and 17 healthy controls were analyzed by flow cytometry. The results indicated that the ratio of pDC in peripheral blood mononuclear cells (PBMNC) was (0.91 ± 064)% in new diagnosed group, which was significantly higher than that in remission group (0.39 ± 0.11)% and control group (0.29 ± 0.13)% (P < 0.01), while this ratio of pDC in remission group was higher than that in control group (P < 0.05). The ratio of mDC in PBMNC was (0.21 ± 0.20)% in new diagnosed group and (0.34 ± 0.21)% in remission group respectively, there was no statistical difference as compared with control group (0.29 ± 0.09)% (P > 0.05). The ratio of pDC to mDC in new diagnosed group was 6.75 ± 7.11, which was significantly higher than that in remission group (1.55 ± 0.93) and control group (1.07 ± 0.43, P < 0.01), there was no statistical difference between the ratio of remission group and control group (P > 0.05). The ratio of pDC in PBMNC of IRP group negatively correlated to ratio of Th1/Th2 (r = -0.347, P < 0.05), and positively correlated to the ratio of auto-antibody on membrane of BMMNC (r = 0.606, P < 0.05) and to the quantity of CD5(+)B cells (r = 0.709, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.381, P < 0.01) and platelets (r = -0.343, P < 0.01). The ratio of mDC in PBMNC positively correlated to the ratio of Th1/Th2 (r = 0.595, P < 0.05) and the level of hemoglobin (r = 0.292, P < 0.05). The ratio of pDC/mDC negatively correlated to ratio of Th1/Th2 (r = -0.395, P < 0.05), it positively correlated to the level of antibody on membrane of BMMNC (r = 0.421, P < 0.05) and the quantity of CD5(+)B cells (r = 0.423, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.304, P < 0.05) and platelets (r = -0.287, P < 0.05). It is concluded that the quantity of pDC in peripheral blood of IRP patients increases, which may be related to the immunopathogenesis of IRP.
Adolescent ; Adult ; Blood Cell Count ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; cytology ; immunology ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; immunology ; Young Adult

Background The nocturnal nondipping and elevated morning blood pressure (BP) in patients with obstructive sleep apnea syndrome (OSAS) have not yet been well investigated in Chinese patients.This study aimed to describe the BP profile,and to elucidate the relationships between daytime BP and nighttime BP,and between evening BP and morning BP in patients with OSAS.Methods Twenty teaching hospital sleep centers in China were organized by the Chinese Medical Association to participate in this study and 2297 patients were recruited between January 2004 and April 2006.BP assessments were made at four time points (daytime,evening,nighttime and morning) and polysomnography (PSG) was performed and subjects were classified into four groups by their apnea-hypopnea index (AHI):control,n=213 with AHI＜5; mild,n=420 with AHI ≥5 and＜15; moderate,n=460 with AHI≥15 and＜30; and severe,n=1204 with AHI ＞30.SPSS 11.5 software package was used for statistical analysis and figure drawing.Results All the average daytime,nighttime,evening and morning BPs were positively correlated with AHI and negatively correlated with nadir nocturnal oxygen saturation.The ralios of nighttime/daytime and morning/evening average BP were positively correlated with AHI.The ratio of nighttime/daytime systolic BP became a “reversed BP dipping” pattern until the classification reached severe,while the ratio of nighttime/daytime diastolic BP became reversed at moderate.Similarly,the ratio of morning/evening diastolic BP becomes reversed even at mild.Conclusions OSAS may result in higher BP levels at all four time points.The ratios of nighttime/daytime and morning/evening BP increase with increased AHI.The increasing of diastolic BP,which is inclined to rise more quickly,is not parallel with increasing systolic BP.

BACKGROUNDEpidemiologic studies have shown an independent and definite association between obstructive sleep apnea (OSA) and hypertension. This study aimed to define the association between daytime blood pressure and severity of OSA in Chinese population in mainland of China.

METHODSTwenty university hospital sleep centers in mainland of China were invited by the Chinese Medical Association (CMA) to participate in this epidemiologic study and 2297 consecutive patients (aged 18 - 85 years; 1981 males and 316 females) referred to these twenty sleep centers for evaluation of OSA between January 2004 and April 2006 were prospectively enrolled. Nocturnal polysomnography was performed in each patient, and disease severity was assessed based on the apneahypopnea index (AHI). These patients were classfied into four groups: nonapneic control (control, n = 257) with AHI < or = 5 episodes/hour; mild sleep apnea (mild, n = 402) with AHI > 5 and < or = 15 episodes/hour; moderate sleep apnea (moderate, n = 460) with AHI > 15 and < or = 30 episodes/hour and severe sleep apnea (severe, n = 1178) with AHI > 30 episodes/hour. Daytime blood pressure measurements were performed under standardized conditions in each patient at 10 a.m. in office on the day of referring to sleep centers for getting average value. All the patients were requested to quit medications related to blood pressure for three days before the day of assessing.

RESULTSBoth daytime systolic blood pressure and diastolic blood pressure values were significantly related to AHI positively (r = 0.201 and 0.276, respectively; both P values < 0.001) and to nadir nocturnal oxygen saturation negatively (r = -0.215 and -0.277, respectively; both P values < 0.001), which were the parameters of OSA severity. In two special designed mean plots, means of daytime systolic and diastolic blood pressure increased gradually with increasing AHI. Beyond AHI of 61 - 65, this increasing trend reached a plateau.

CONCLUSIONSThe results showed that OSA severity was associated with daytime blood pressure until AHI of 61 - 65, providing evidence for early OSA management, especially in OSA patients with concomitant hypertension.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Pressure ; physiology ; China ; Female ; Humans ; Male ; Middle Aged ; Sleep Apnea, Obstructive ; pathology ; Young Adult

OBJECTIVESTo investigate the expression of P120 catenin in pancreatic carcinoma and to explore the association between P120 catenin gene polymorphism at T755G position and pancreatic carcinoma.

METHODSThe expression of P120 catenin in 52 cases of pancreatic carcinoma and normal pancreatic tissues on the mRNA and protein levels were evaluated by RT-PCR and Western Blot methods respectively. P120 catenin gene polymorphism at T755G position of in 52 patients and 60 healthy controls were examined by PCR-restriction fragment length polymorphism (PCR-RFLP) technique.

RESULTSThe mRNA and protein expressions of P120 catenin in pancreatic carcinoma tissues were significantly lower than normal pancreatic tissues (P=0.000, P=0.002). Reduced expression of P120 catenin mRNA was significantly correlated with differentiated (P=0.033), lymph node metastasis (P=0.004), vascular invasion (P=0.022), and pTNM stage (P=0.003). Additionally, there were significant difference of P120 catenin gene polymorphism genotypes and alleles at T755G position between patients and healthy controls (P=0.008, P=0.016). The GG genotype of P120 catenin gene was associated with higher risk of incidence for pancreatic carcinoma compared with the TT genotype (OR=2.765, 95%CI=1.312-3.958).

CONCLUSIONSThe reduced expressions of both P120 catenin mRNA and protein in pancreatic carcinoma suggest its association with pancreatic carcinoma development. Polymorphism of P120 catenin gene at T755G situation might be a risk factor for pancreatic carcinoma, and it may be used to diagnosis and prevent pancreatic carcinoma early.

Case-Control Studies ; Catenins ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; genetics ; metabolism ; Polymorphism, Genetic

OBJECTIVETo investigate the types and therapies of malignancies in renal allograft recipients.

METHODSWe retrospectively analyzed the occurrence, types, and therapies of malignancies in 498 renal allograft recipients who had received operations in Peking Union Medical College Hospital from May 1986 to October 2008.

RESULTSAmong 498 renal allograft recipients, 18 patients (3.6% ) were diagnosed with malignancies, which included bladder cancer (n = 5), renal pyloric cancer or ureteric cancer (n = 4), leukemia or lymphoma (n = 3), hepatic cancer (n = 2), skin cancer, rectum carcinoma, pulmonary carcinoma and thymoma (n = 1 each). Surgical operations were performed in 10 cases, 6 of whom survived with normal renal function and had no rejection of transplanted kidneys. Three patients with bladder cancer and one patient with ureteric cancer experienced recurrences 7 to 15 months after operations; among them one bladder cancer patient died. One hepatic carcinoma patient died of pulmonary metastasis 8 months after operation. One non-Hodgkin's lymphoma patient died 11 months after chemotherapy. Five cases with advanced unresectable malignancies died 8 to 17 months after the diagnosis.

CONCLUSIONSThe incidences of malignancies, especially urological epithelial carcinoma, are high in renal allograft recipients. Radical surgery of the solid malignancies is a preferred option.

Adult ; Aged ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Neoplasms ; epidemiology ; therapy ; Postoperative Complications ; epidemiology ; therapy ; Retrospective Studies

OBJECTIVETo study the inducible expression of hypoxia-inducible factor 1alpha (HIF-1alpha) on the proliferation and invasion property of HepG2 cells under normoxia in vitro.

METHODSConstructed the HepG2(Tet-on-HIF-1alpha) cell line which could induce the expression of HIF-1alpha by doxycycline; Under normoxia in vitro, MTT assay was used to observe the proliferative and adhesive activity of cells, and the invasive activity was determined by transwell cell culture chamber method.

RESULTSUnder normoxia, the HIF-1alpha mRNA and protein of HepG2(Tet-on-HIF-1alpha) cells could be induced up to (5.899 +/- 2.176) and (2.179 +/- 0.742) folds by doxycycline (1 microg/ml); There were no difference of A(490 nm) between the Dox(+)and Dox(-) group in experiment detecting the proliferation activity (P > 0.05); But in adhesive experiment, the A(490 nm) of Dox (+) group was 0.662 +/- 0.058, higher than the Dox(-) group 0.526 +/- 0.808 (P = 0.008); The invasive cell number of Dox(+) group was 37.611 +/- 8.424, but in the Dox(-) group, the number was 25.333 +/- 8.117 (P < 0.01).

CONCLUSIONSUnder normoxia in vitro, the Tet-on gene regulate system could increase the HIF-1alpha protein by inducing the HIF-1alpha mRNA; HIF-1alpha has no influence with the proliferation activity, but it could enhance the adhesive and invasive properties of HepG2 cells.

Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cell Survival ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; physiology ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Invasiveness ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction