The CrdS protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to explore the regulatory mechanism in the acidic adaptation of Helicobacter pylori (H. pylori). The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS protein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm-T-hp1364 and the prokaryotic expression plasmid pQE30-hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XL1 blue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.
Bacterial Proteins ; biosynthesis ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Helicobacter pylori ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

Objective To evaluate the therapeutic effects,acute and late side-effects of cisplatin plus 5-fluorouracil (PF) schemed chronomodulated chemotherapy combined with concurrent intensitymodulated radiotherapy(IMRT) for nasopharyngeal carcinoma.Methods A total of 48 nasopharyngeal carcinoma patients treated with chronomodulated chemotherapy plus concurrent IMRT were retrospectively studied.The radiation doses prescribed to the gross target volume GTVnx of nasopharyngeal,to the positive neck lymphnodes GTVnd,to the clinical target volume CTV1,and to CTV2 were 73.92-77.88,69.96,60.06-66.00 and 50.96-56.00 Gy respectively.The Kaplan-Meier survival analysis was used to calculate the local-regional progression-free rate.The acute and late side-effects were graded according to RTOG radiation morbidity scoring criteria.Results The complete remission (CR) rate was 41.6％(20/48) ; the partial remission (PR) rate was 47.9％ (23/48) ; the stable disease (SD) rate was 4.2％(2/48) ; and the local tumor control rate was 89.6％.Most of the patients had grade 1 to 2 acute side effects and grade 0 to 1 late side-effects.No grade 4 acute and late toxicity was observed.1,2,4-year survival rates were 93.8％,79.2％ and 64.5％,respectively.The DVH analysis showed that IMRT improved the total dose and dose per-fraction to the target volume with considerable dose reduction of the normal structures at the same time.Conclusions PF chronomodulated chemotherapy plus concurrent intensity-modulated radiotherapy for NPC could improve therapeutic effects with satisfactory tolerance of the side effects.

Objective To evaluate the value of real-time three-dimensional echocardiography(RT-3DE)in diagnosing congenital tricuspid valve anomaly.Methods Eighteen patients with congenital tricuspid valve anomaly were studied by RT-3DE,the spatial framework and neighboring structures of the tricuspid valve were analyzed and compared with result of the operation.Results The anomaly of tricuspid,chorda tendineae,the papillary muscle and their connection with neighboring structures could be displayed clearly from different directions.The diagnose accordance rate of RT-3DE was 83%.Conclusions RT-3DE may provide more information on congenital tricuspid valve anomaly than 2DE.

ObjectiveTo investigate the delineation of gross tumor volume (GTV) in locally advanced nasopharyngeal carcinoma (LANC) according to imageological changes before and after induction chemotherapy (IC) in order to decrease high dose area and protect normal tissue better.MethodsBetween Mar 2010 to Jan 2011,11 patients with LANC were enrolled and treated with TPF regimen followed by intensity-modulated radiotherapy (IMRT) with concurrent chemotherapy,target volumes were delineated based on fused CT imaging before and after IC following project determination.Tumor target volumes after and before IC were respectively delineated according to imaging tumor residues and were overlaid by CTVnx in order to ensure radical doses for the imaging tumor volume before IC,the resulting differences of tumor target volumes of IC before and after were measured and analyzed by paired t-test.ResultsBefore and after IC,the average volumes of GTVnx were respectively 44.72 cm3 and 28.87 ( t =3.89,P =0.003 ),the average volumes of GTVnd were respectively 32.76 cm3 and 19.82 cm3 ( t =2.47,P =0.033 ),the volumes of maximum dose area in brainstem and spinal cord as well as eyeball decreased ( t =2.93-4.59,all P ＜0.05).ConclusionsLANC treated by 3 cycle TPF regimen followed by IMRT with concurrent chemotherapy showes significant shrinkage of tumor volume.The volume of high dose region which caused by normally recovered tissues were decreased by re-delineation of target volume in brainstem and spinal cord as well as eyeball of CT images after IC.

Objective To investigate the dosimetric influence of pure carbon fiber treatment tabletop of Elekta Precise new linear accelerator in radiotherapy.Methods Surface-axis distance (SAD) technology was employed for the measurement.Two groups of fields were set and both of them were SAD opposed portals ( one of them went through the tabletop,while the other did not).A PTW electrometer and a 0.6 cm3 Farmer ionization chamber were utilized for comparison measurement.Then dose attenuation of the main table board,extended body board,the extended board for head,neck and shoulders,and the joints of these boards were calculated.Results Under the energy of 6 MV,the dose attenuations of the following locations were:1.4％ - 7.2％ at the main treatment table board; 2.8％ - 38.7％,1.4％ -30.1％,1.5％ -20.8％ and 1.4％ - 11.2％,respectively at distances of 1,4,7 and 8 cm from the joint of the main table board ;0.5％ - 5.0％ at the extended body board; 4.7％ - 15.4％ at distance of 1cm from the joint of the extended body board; 0.5％ -3.3％ at the neck position of the extended board for head,neck and shoulders; 5.3％ - 16.7％ at the shoulder positions; and 6.8％ -30.4％ at the joint between the extended boards and the main table board.Conclusions The dose attenuations of the new linear accelerator pure carbon fiber treatment tabletop vary at different locations. Considerable higher attenuations are observed at the table board joints than other locations.

Objective To evaluate the application value of three-dimensional speckle tracking imaging (3D-STI) in quantitatively evaluating the left ventricular global strain in recipients within 3 months after renal transplantation. Methods Clinical data including blood pressure, serum creatinine and tacrolimus blood concentration of 34 renal transplant recipients were collected before operation, 7 d, 1 month and 3 months after operation, respectively. Meanwhile, conventional echocardiography and 3D-STI examination were performed. Echocardiographic parameters [left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and left ventricular ejection fraction (LVEF)] and 3D-STI parameters [left ventricular global peak longitudinal strain (GPLS), global peak circumferential strain (GPCS), global peak radial strain (GPRS) and global peak area strain (GPAS)] of recipients were collected. The changes of these parameters before operation, 7 d, 1 month and 3 months after operation were statistically compared. The changing characteristic and application value of 3D-STI in quantitatively evaluating the left ventricular global strain in recipients within 3 months after renal transplantation were evaluated. Results LVEF and GPCS did not significantly differ at different time points (all P > 0.05), whereas LVEDV, LVESV, GPLS, GPAS and GPRS significantly differed at different time points from preoperative to within postoperative 3 months (all P < 0.001). GPLS, GPAS and GPRS trended to decline within postoperative 1 month, and slightly increased at 3 months after operation, which was still lower than the preoperative levels. Conclusions Application of 3D-STI may sensitively detect the changes of left ventricular global strain in recipients after renal transplantation when no significant variations are observed in postoperative LVEF. Compared with conventional echocardiography, 3D-STI may more accurately evaluate the changes of left ventricular global strain in recipients after renal transplantation.

Objective To evaluate the differences of toxicities,therapeutic efficacy and immune function between induction chemotherapy followed by sinusoidal chrono-modulated infusion and flat intermittent infusion of cisplatin (DDP)with intensity-modulated radiotherapy (IMRT) in patients with locoregionally advanced nasopharyngeal carcinoma (NPC).Methods Seventy patients with biopsydiagnosed stages Ⅲ and Ⅳ B NPC (according to the 2010 UICC staging system) were treated with two-cycle induction chemotherapy before chemoradiotherapy in Guizhou Cancer Hospital.The TPF chemotherapy regimen was administered as follows:The TXT and DDP with the dose of 75 mg/m2 was carried out by bolus infusing for the first day,the 5-FU with 750 mg · m-2 · d-1 was carried out by continuous intravenous pumping for the first day to fifth day(120 h).The induction chemotherapy was 21 days per cycle,for two cycles.After that all patients were randomly treated with 2-3 cycles of sinusoidal chronomodulated infusion or flat intermittent constant rate infusion of DDP with IMRT.Using a multi-channel programmed pump,the patients were given 12 h continuous infusions of DDP (100 mg/m2) for day one,repeated every 3 weeks for 2-3 cycles.DDP was administered from 10:00 am to 10:00 pm.Concurrent radiotherapy regimen was administered as follows:GTVnx 69.96-73.92 Gy/33 f,PTVnx 69.96 Gy/33 f,PTVnd 69.96 Gy/33 f,PTV1 60.06 Gy/33 f,PTV2 50.96 Gy/28 f.Results The main toxicities of chemoradiotherapy in the group of sinusoidal chrono-modulated infusion were bone marrow suppression:leukocytes,and then nausea,oral mucositis and hemoglobin.The main toxicities of chemoradiotherapy in the group of flat intermittent constant rate infusion were bone marrow suppression:hemoglobin,leukocytes,and then nausea,oral mucositis.No significant differences were observed for toxicities(P ＞ 0.05).After concurrent chemoradiotherapy,the complete response rate (CR),partial response rate (PR),stable disease rate(SD),progressive disease rate (PD) and overall response rate (ORR) were 11.4％,85.7％,2.9％,0 and 97.1％ in the group of sinusoidal chrono-modulated infusion.The CR,PR,SD,PD,ORR in the group of flat intermittent constant rate infusion were 22.9％,74.2％,2.9％,0,97.1％,respectively.However,there was no significant differences of effect in the two Arms (P ＞ 0.05).For sinusoidal ehrono-modulated infusion and flat intermittent infusion group,the 2-year overall survival(OS) were 82.9％ and 94.3％ respectively,the 2-year progression-free survival(PFS) were 77.1％,91.4％ respectively,and the 2-year distant metastasis free survival (DMFS) were 82.9％,91.4％ respectively.The value of CD3 + in the group of sinusoidal chrono-modulated infusion was higher than the group of flat intermittent constant rate infusion after therapy (Z =3.254,P ＜ 0.05).The value of CD4 +,CD8 +,CD16 + CD56 +,CD19 +,and CD4 +/CD8 + had no differences in two Arms (P ＞ 0.05).Conclusions No significance differences on the toxicities,therapeutic efficacy and survival were observed between the two groups,but immune function might be improved in the sinusoidal chrono-modulated infusion group.

Objective To compare the complications and outcomes of urgent?start peritoneal dialysis (PD) and hemodialysis (HD) in end?stage renal disease (ESRD) patients, and explore the safety and effectiveness of PD which was as an urgent?start dialysis modality in ESRD patients. Methods All patients for urgent?start dialysis, who initiated dialysis without a long?term dialysis access or had the long?term dialysis access under 30 days in Renji Hospital from January 1st 2013 to December 31st 2014, were enrolled. According to the dialysis modalities, patients were divided into PD group and HD group. Participants were followed up until death, transferred to other centers, lost of follow up or January 1st 2016. Dialysis?related complications within 30 days of implantation, complications of reimplantation and the occurrence of bacteremia between two groups were compared, and their survival rates were tested by Kaplan?Meier curves. Results Among 178 patients in this study, there were 96 (53.9%) patients in PD group and 82 (46.1%) patients in HD group. Compared with those of HD group, patients of PD group presented more cardiovascular disease [21(21.9%) vs 8(9.8%), P=0.029], higher serum potassium [(4.5±0.8) mmol/L vs (4.3±0.8) mmol/L, P=0.038], but less heart failure (NYHA Ⅲ?Ⅳ) [26(30.2%) vs 40 (48.8%), P=0.014], lower brain natriuretic peptide (BNP) [328.5 (129.5, 776.8) ng/L vs 503.5(206.0, 1430.0) ng/L, P=0.008], higher hemoglobin [(81.5 ± 17.7) g/L vs (75.3 ± 22.5) g/L, P=0.039], higher serum albumin (33.5±5.7) g/L vs (31.3±6.7) g/L, P=0.022] and higher serum pre?albumin (304.5±78.0) mg/L vs (257.0 ± 86.1) mg/L, P<0.001]. PD group presented less dialysis?related complications [5 (5.2%) vs 20(24.4%), P<0.001], less dialysis?related complications requiring reimplantation [1(1.0%) vs 20(24.4%), P<0.001] and less bacteraemia [3(3.1%) vs 11(13.4%), P=0.011]. The 3?, 6?and 12?month patient survival rates of PD and HD group were 97.9% vs 98.4%, 97.9% vs 98.4%, and 92.1%vs 93.0% respectively, and no significant difference was found (Log ? rank=0.004, P=0.947). Conclusions Patients with urgent?start PD have less complications within 30 days of implantation and occurrence of bacteremia than patients with urgent?start HD, and the same survival rates. PD may be a feasible and safe urgent?start dialysis modality for ESRD patients.

Objective To systematically evaluate the optimal dose of early enteral nutrition support in critically ill patients.Methods Systematic search database including PubMed,Web of science,Scopus,CINAHL,CBM,CNKI.RCTs about early enteral nutrition dose selections in critically ill patients were chosen according to include and exclude criteria by two researchers independently.Cochrane system evaluation manual bias risk assessment was used to evaluate quality of literature.RevMan5.3 Meta analysis software was used to analyze the data.Results A total of 1 571 literatures were retrieved and 8 RCT studies were included,2 713 subjects in total.Meta analysis results showed that there were statistically significant differences in mechanical ventilation time,incidence of diarrhea,and utilization rate of gastro dynamic drugs between trophic feeding and full feeding (P＜0.05).There were no statistically significant differences in mortality,length of stay,incidence of nosocomial infections,reflux,vomiting,constipation,etc.(P＞0.05).Conclusions Trophic feeding has familiar effects on mortality,length of hospital stay compared to full feeding,but it can help to shorten ICU mechanical ventilation time,improve the gastrointestinal tolerability.

Objective:To investigate the effect of Kouyanqing granule combined with ornidazole on the expression of high mobility group protein B1 (HMGB1), tumor necrosis factor-α (TNF-α), interleukin-17 (IL-17) and interleukin-23 (IL-23) in patients with chronic periodontitis.Methods:From October 2018 to October 2019, 162 patients with chronic periodontitis admitted to the First People's Hospital of Xiaoshan District were divided into observation group (81 cases) and control group (81 cases) according to the random digital table method.The control group was treated with ornidazole, and the observation group was treated with Kouyanqing granules on the basis of the control group.The course of treatment in both two groups was 4 weeks.The changes of plaque index (PLI), probing depth of periodontal pocket (PD), bleeding index (BI), HMGB1, TNF-α, IL-17 and IL-23 and adverse reactions were compared before and after treatment.Results:The total effective rate of the observation group (91.36%) was higher than that of the control group (77.78%) (χ 2=5.723, P<0.05). The PLI(0.93±0.16), PD[(3.15±0.37)mm] and BI(1.35±0.24) in the observation group were lower than those in the control group [(1.42±0.24), (4.28±0.45)mm and (2.18±0.28)] ( t=15.289, 17.457, 20.256, all P<0.05). The levels of HMGB1 [(0.78±0.13)μg/L], TNF-α[(3.10±0.32)μg/L], IL-17[(368.19±24.31)ng/L] and IL-23[(7.63±0.89)ng/L] in the observation group were lower than those in the control group [(0.96±0.10)μg/L, (4.34±0.46)μg/L, (457.32±21.35)ng/L and (10.32±1.25)ng/L] ( t=9.877, 19.916, 24.793, 15.777, all P<0.05). The incidence of adverse reactions in the observation group (6.17%) was lower than that in the control group (17.28%) (χ 2=4.830, P<0.05). Conclusion:Kouyanqing granule combined with ornidazole is effective in the treatment of chronic periodontitis.It can reduce the levels of HMGB1, TNF-α, IL-17 and IL-23, and has few adverse reactions.