The DegP protein, functioning as both chaperone and protease, plays a critical role in degrading and removing denatured or damaged proteins in the cellular envelope during heat shock and other stresses. So far, several proteins have been identified as its natural targets. A carboxyle-terminal peptide derived from the PapG pilus, one of the in vivo substrates for DegP, has been shown to activate the protease. Nevertheless, neither the details nor the physiological implications of such activation have been studied. The evidence that DegP undergoes conformational changes upon binding the peptide derived from C-terminal sequence of pilus subunit PapG has been presented. It demonstrated that upon binding this peptide, detectable changes can be observed for both secondary and tertiary structures of DegP, as examined by CD spectroscopy. Gel filtration and dynamic light scattering analysis also revealed that the size of DegP becomes smaller to a minor extent. Moreover, both the hydrophobic surfaces and catalytic sites of DegP were found to expose slightly in the presence of the peptide. Upon peptide binding, a less flexible and more rigid conformation of DegP was obtained as analyzed by fluorescence anisotropy. The physiological implications of these observations for DegP are discussed.

Objective To explore the efficacy and safety of sequential telbivudine ( LDT) therapy following interferon-α( IFN-α) in hepatitis B e antigen (HBeAg) positive chronic hepatitis B patients.Methods 94 chronic hepatitis B patients in our hospital hepatitis clinic archives from August 2008 to September 2013 were divided into two groups, 45 patients treated with LDT following IFN-αdue to unsatisfactory response to peginterferon-α-2a (peg-IFN-α-2a)treatment in the sequential group and 49 patients treated with LDT initially in the controlled group.In the sequential group, 22 patients failed to respond to IFN-αtreatment, 4 patients obtained IFN-resistance, 6 patients developed severe side effects, 1 patients had pre-C mutation and 12 patients switched to LDT due to economic factors or fertility requirement.The biochemical response and virological response, virological breakthrough and serological response were observed.Chi-square test was adapted for data analysis.Results After 32.35 months of LDT therapy, 88.9%(40/45) patients had with HBV-DNA <100 IU/mL in the experiment group compared with 44.9%(22/49) patients in the control group (P<0.05).The rates of HBeAg loss and HBeAg seroconversion in the experiment group were 53.3%(24/45) and 51.1%(23/45) respectively, which were significantly higher than that of the control group, 24.5%(12/49) and 24.5% (12/49) (P<0.05).6.7% (3/45) patients developed virological breakthrough in the sequential group compared with 34.7%(17/49) in the control group (P<0.05).Above results suggested the synergistic antiviral activity between peg-IFN-α-2a and LDT.Conclusion Sequential LDT therapy following peg-IFN-α-2a is better efficacy and safety compared with treating patients with LDT alone initially.

Adolescent ; Adult ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; diagnosis ; drug therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thymidine ; analogs & derivatives ; therapeutic use ; Treatment Outcome ; Young Adult

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Objective To investigate the coverage of a recombinant protein vaccine based on pneumococcal surface protein A (PspA) from both family 1 and family 2.Methods One hundred and fifty-nine Streptococcus pneumoniae strains, including 47 invasive strains, were isolated from children in Nanjing Children′s Hospital.Cell lysates were prepared and reacted with three antibodies recognizing PspA -RX1, PspA-3296 and PspA-5668 for PspA typing by ELISA .Results Among 47 invasive isolates of 9 different serotypes, 10.7%were PspA family 1 and 89.3%were PspA family 2.Among all of 159 clinical isolates, 10.1% were identified as PspA family 1, 88.0%were family 2, while 1.9%of strains could not be typed by ELISA and PCR assays .None of strains belonged to PspA family 3.Conclusion The recombinant pro-tein vaccine based on PspA from both family 1 and family 2 has a broad coverage among clinical isolates and is potentially protective against both invasive and non-invasive pneumococcal diseases .

Objective To explore the influence of G4 cyberknife treatment of large hepatocellular car-cinoma on liver function,and to evaluate its treatment safety.Methods Sixty-three large liver cancer patients treated with routine G4 cyberknife treatment were retrospectively analyzed,and then statistical analysis of the difference in liver function before and after treatment was conducted.Results After G4 cyberknife treatment of 1 2 months,the levels of ALT,ALP,TBIL,PA were respectively 23.00 U /L,1 1 1 .00 U /L,1 3.70 μmol/L, (81 .87 ±1 3.94)%.Compared with the levels before treatment [28.00 U /L,32.00 U /L,1 1 .30 μmol/L, (86.07 ±1 4.07)%],there were no signi-ficant differences found (Z =-1 .677,P =0.094;Z =-0.504, P =0.61 4;Z =-1 .945,P =0.053;t =1 .271 ,P =0.21 3).The level of ambumin was (34.84 ±4.75)g/L at 1 2 months after treatment,which decreased and the difference compared with the level before treatment [(37.45 ±4.1 4)g/L]was significant (t =3.357,P =0.002).The Child-Pugh grade was 5.80 ±1 .1 7 respectively at the time points of 1 2 months after treatment,and no significant difference was found compared with the Child-Pugh grade before treatment (5.48 ±0.81 ,t =-1 .668,P =0.1 06).Conclusion G4 cyberknife treatment does not cause liver injury.It is safe and reliable in large liver cancer treatment.So,it is worth widely clinical popularizing.

Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid (SAHA) on ovarian carcinoma. Methods (1)Two groups of ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM) were exposed to SAHA (1, 3, 5 and 7μmol/L SAHA,group 1-group 4). CCK-8 method was employed to eval-uate the inhibitory effects of SAHA.(2)Ovarian cancer cell lines treated with SAHA (2 or 5μmol/L SAHA) were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis.(3) Reverse transcription polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the mRNA and pro-tein expression levels of phenotypic correlation factor. Results (1)After 48 h of SAHA treatment,the OD value of SKOV3, SKOV3/DDP,and HO8910 showed a trend of gradually reduce (P<0.05).(2)The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group (P<0.05). Compared with control group, after 48 h of SAHA treat-ment,S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased;G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups (P<0.05).(3)The expression levels of CyclinB1 and Cdc2 (p34) mRNA were significant-ly lower in SAHA 1 and 2 groups than those of control group,while the expression levels of Caspase-3,p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,p53 protein were markedly improved,and CyclinB1,Cdc2(p34) protein decreased in SAHA 1-4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone acetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB1 and Cdc2(p34).

OBJECTIVETo illuminate the regulating effect of all-trans retinoic acid (ATRA) on gap junctional intercellular communication (GJIC) and connexin 43 (Cx43) gene expression in glioma cells, which is tissue- and organ-specific.

METHODRat C6 glioma cells were exposed to ATRA at a concentration of 1, 10, 100 micromol/L and the GJIC function of the cells was examined with scrape-loading dye transfer assay 24 hours, 48 hours and 72 hours after ATRA treatment. The effect of ATRA on Cx43 gene expression was measured with semiquantitative reverse transcription polymerase chain reaction (RT-PCR) 24 hours after ATRA exposure.

RESULTSThe GJIC function of C6 glioma cells was significantly increased by ATRA at each concentration applied. The dye passed 4 to 5 rows of cells from the scraping edge in ATRA treated cells, but only 1 or 2 rows in the control. The augment effect was observed 24 hours after each concentration ATRA treatment, and lasted till 72 hours after treatment with 1 micromol/L and 10 micromol/L ATRA. Forty-eight hours after exposed to 100 micromol/L ATRA, the enhancement of GJIC was less obvious. There was no significant increase induced by ATRA on the transcription of Cx43 gene, as demonstrated by semiquantitative RT-PCR.

CONCLUSIONATRA turned out to be a potent enhancer on GJIC function in C6 glioma cells, andthe enhancement effect was most probable at post-transcriptional level.

Animals ; Antineoplastic Agents ; pharmacology ; Brain Neoplasms ; metabolism ; pathology ; Connexin 43 ; biosynthesis ; genetics ; Gap Junctions ; physiology ; Gene Expression ; Glioma ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rats ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

OBJECTIVE:To observe analgesia,sedation effects and safety of flurbiprofen axetil combined with hydromorphone for postoperative patient-controlled intravenous analgesia (PCIA) after orthopedics surgery. METHODS:Totally 90 patients with combined spinal epidural anesthesia underwent lower limb surgery were selected from anesthesology department in the Affiliated Hospital of Chengde Medical College during May 2016-Jan. 2018. They were divided into SF group,H group and KH group according random number table,with 30 cases in each group. The postoperative PCIA pump drug liquid formula of SF group included Sufentanil citrate injection 2-3 μ g/kg+Tropisetron hydrochloride for injection 10 mg+0.9% Sodium chloride injection diluted to 100 mL;that of H group included Hydromorphone hydrochloride injection 0.12 mg/kg+Tropisetron hydrochloride for injection 10 mg+0.9% Sodium chloride injection diluted to 100 mL;that of KH group included Hydromorphone hydrochloride injection 0.12 mg/kg+Flurbiprofen axetil injection 50 mg+Tropisetron hydrochloride for injection 10 mg+0.9% Sodium chloride injection diluted to 100 mL. The operation time, intraoperative medication (epidural application frequency of additional ropivacaine,frequency of ephedrine and atropine),effective pressing times of analgesic pump and the analgesic effect of PCIA were observed in 3 groups. VAS score and Ramsay sedation score were observed 2,6,12,24,48 h after surgery. The hospital anxiety and depression scale (HAD) score,profile of mood states (POMS) score and the occurrence of ADR were observed before and after surgery. RESULTS:There was no statistical significance in operation time,epidural application frequency of additional ropivacaine or frequency of ephedrine and atropine among 3 groups (P＞0.05). The effective pressing times of analgesic pump in KH group were significantly lower than SF group and H group. The proportion of patients with excellent and good anesthesia effect in KH group was significantly higher than SF group and H group (P＜0.05);there was no statistical significance between SF group and H group(P＞0.05). VAS score of 3 groups 48 h after surgery were significantly lower than 6, 12,24 h after surgery;that of KH group was significantly lower than SF group and H group(P＜0.05);there was no statistical significance between SF group and H group(P＞0.05). There was no statistical significance in Ramsay score among 3 groups at different time points(P＞0.05). Before surgery,there was no statistical significance in HAD score or POMS score among 3 groups (P＞0.05). After surgery,HAD score and POMS score of KH group and H group were significantly lower than before surgery and SF group(P＜0.05);there was no statistical significance in KH group and H group,before and after surgery in SF group(P＞0.05). No vomiting,respiratory depression,pruritus and digestive tract bleeding were observed in 3 groups. The incidence of dizziness and nausea in H group and KH group were significantly lower than SF group (P＜0.05);there was no statistical significance between KH group and H group(P＞0.05). CONCLUSIONS:The flurbiprofen axetil combined with hydromorphone show good analgesic and sedative effect for PCIA after orthopedics operation,and can significantly improve emotion and mood of patients with good safety.

Objective To validate a chemical shift-encoded MRI(CSE-MRI)water-fat imaging for quantifying vertebral marrow fat content using MRS as the reference standard.Methods MRS and CSE-MRI were performed to calculate proton density fat fraction(PDFF) in 83 subjects,including 41 normal bone mass,26 osteopenia and 16 osteoporosis.Eight participants were scanned three times with repositioning to assess the repeatability of CSE-MRI PDFF measurements.Agreements of intra-observer and inter-observer were evaluated by intraclass correlation coefficient(ICC).Linear regression,Bland-Altman 95% limit of agreement and Lin's concordance correlation coefficient were calculated.Results The repeatability for CSE-MRI PDFF measurements expressed as absolute precision error was 1.45%.PDFF was 62.1%±11.1% by MRS and 60.4%±10.1% by CSE-MRI in 83 subjects.There were significant differences in PDFF among the normal bone mass,osteopenia and osteoporosis groups after adjusting for age,years since menopause and body mass index (all P<0.001).The intra-and inter-rater reliability for duplicate measurements at CSE-MRI PDFF were more than 0.993.Pearson correlation coefficient was 0.979 and Lin's concordance correlation coefficient was 0.962.All data points calculated using the Bland-Altman method were within the limits of agreement.Inverse associations were observed between BMD (r=-0.560--0.710)and CSE-MRI-based PDFF,and between BMD (r=-0.539--0.706)and MRS-based PDFF in various groups.Conclusion CSE-MRI with multiple lipids peak model and T2?-correction is equally accurate in characterizing marrow fat content as MRS.