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 investigate the effect of miR-15b and miR-16 on the expression of vascular endothelial growth factor (VEGF) protein in human embryonic lung fibroblast (HLF) cells under hyperoxia.Methods The expression level of miR-15b and miR-16 was up-regulated and down-regulated in HLF cells by transfection technology, respectively.The expression of VEGF protein in HLF cells was assessed by Western blot.Furthermore, under hyperoxia exposure in vitro, the expression of miR-15b, miR-16 and VEGF protein in HLF cells was analyzed.Results Up-regulation of miR-15b and miR-16 suppressed VEGF protein expression, while down-regulated miR-15b and miR-16 promoted VEGF protein expression.In addition, hyperoxia exposure induced up-regulation of miR-15b and miR-16, but down-regulation of VEGF protein in HLF cells.Conclusions Hyperoxia exposure may up-regulate the expression level of miR-15b and miR-16, but suppress VEGF protein expression.These may contribute to the development of bronchopulmonary dysplasia.

Objective To study and evaluate the MRI findings and their diagnostic value for breast carcinoma.Methods Preoperative contrast-enhanced MR imaging and mammography were performed in 12 patients who underwent surgery and were histopathologically proved with breast carcinoma.MR imaging findings were compared with clearly mammography.Results Twenty-one breast lesions in 12 cases were detected with MR imaging.Seventeen of the twenty-one lesions were detected with mammography.Contrast-enhanced MR imaging was superior to mammography to demonstrate lesions in these dense breasts.It could clearly indicate the extension of posterior breast carcinoma into the pectoralis major muscle and the metastases of parasternal lymphatic node,while it was difficult to evaluate with mammography.The "rim enhancement sign" might be valuable to diagnose breast carcinoma.Conclusion The sensitivity and accuracy of MR imaging is superior to that of mammography.Contrast-enhanced imaging of the breast is helpful not only in early diagnosis of breast carcinoma but also in staging breast carcinoma and planning therapy.

Objective To evaluate clinical factors as predictors of radiation pneumonitis(RP)in advanced stage non-small cell lung cancer(NSCLC)patients treated with concurrent radiochemotherapy when gross tumor volume is 70 Gy. Methods Data of 84 patients with histologically proved NSCLC treated with 3DCRT or IMRT were collected. To evaluate the correlation between clinical parameters and radiation pneumonitis(RP). The clinical parameters were considered: pathological type, therapy agents, age,gender, stage, karnofsky performance status(KPS), smoking status, diabetes, chronic obstructive pulmonary disease(COPD). Results The occurrence of grade 1,2 RP was 63%, 33%, respectively. In univariate analysis, diabetes was significantly associated with RP of ≥ grade 1(x2 =4.03, P = 0.045)and ≥grade 2(x2 = 15.59 ,P =0.000). KPS was significantly associated with RP of ≥grade 1(x2 =3.98 ,P = 0.046)and ≥grade 2(x2 = 5.21, P = 0.023). In logistic multivariate analysis, diabetes was significantly associated with RP of ≥grade 1(x2 =5.50,P =0.019)and ≥grade 2(x2 = 12.92,P =0.000). KPS was significantly associated with RP of ≥ grade 1(x2 = 6.29, P = 0.012)and ≥ grade 2(x2 = 6.61, P =0.010). Conclusion The definite statistical significant risk factors of RP are diabetes and KPS.

ObjectiveThis study evaluates the feasibility of intensity-modulated radiation therapy (IMRT) to treat patients with 1 -5 brain metastases from non-small cell lung cancer (NSCLC).Methods 30 IMRT patients with brain metastases for NSCLC studied retrospectively.Whole brain radiotherapy plus three-dimensional conformal radiotherapy (WBRT + 3DCRT) and WBRT plus stereotactic radiotherapy ( WBRT + SRT) plans were generated.Planning target volume ( PTV ) and organs at risk dose were measured and compared by dose volume histogram.Differences were analyzed in the three techniques by Wilcoxon Z -test.Results D99％ of the shoulder ( D99％-D90％ ) from IMRT were higher than from WBRT +3DCRT and WBRT+SRT in all cases.From D15％ of slope (D90％-D10％) to D5％ of tail (D10％ -D1％ ),IMRT were lower than WBRT + 3DCRT and WBRT + SRT ( Z =- 4.72,P =0.000 and Z =- 4.72,P =0.000).D10％ and D5％ of IMRT were (35.1 ±1.42) Gy and (37.7 ±2.91) Gy,WBRT +3DCRT were (36.5±2.86) Gy and ( 39.1 ± 3.56) Gy ;WBRT + SRT were (36.2 ± 2.57) Gy and ( 38.7 ± 3.67) Gy.IMRT vs WBRT+ 3DCRT and WBRT + SRT were significant ( Z=-3.18,-3.18,P=0.001,0.001 and Z=- 4.11,- 3.02,P =0.000,0.002) in 13 patients with 3 - 5 brain metastases.The total mean monitor units were 14756.3,9614.8 and 9043.2 for IMRT,WBRT +3DCRT and WBRT + SRT plans,respectively,with a 38.7％ reduction from IMRT to WBRT + SRT (Z =-4.78,-4.78,P =0.000,0.000).The brain doses around metastases were similar in the three techniques with 1 -2 metastases,but IMRT was the best with 3 -5 metastases.ConclusionsIMRT can advance brain metastases dose and improve the planning target minimum dose and spare the dose around brain metastases.Only IMRT is the best choice for just sparing the dose around brain metastases among 3 -5 brain metastases.

Objective To analyze the clinical features of 35 cases of Kennedy's disease and the correlation be?tween clinical features and CAG repeat size to strengthen the understanding of KD and to avoid misdiagnosis and delayed diagnosis.Methods Clinical data, including clinical signs and symptoms ,serum lipid, serum sex hormone level, electro?myography, the number of CAGs and (amyotrophic lateral sclerosis muscular atrophy,ALS) rating scale were collected from 35 patients genetically diagnosed of Kennedy disease and proceed system analysis. Results Patients with KD were adult onset with the average age of (40.77 ± 8.57) years and the average confirmed course were (8.32 ± 4.17) years. Forty-two point nine percent of the patients had family history. Clinical features included medulla oblongata and spinal muscular atrophy and weakness, limbs tremor, perioral muscles twitch and endocrine function and metabolic disorders in some cases. Creatine kinase, triglyceride, low density lipoprotein, follicle estrogen and prolactin were significantly in?creased compared to healthy adults (P:0.000,0.018,0.000,0.000,0.003). The number of CAG repeat was negatively correlated with the onset age (r=-0.549, P=0.001) but not associated with the illness severity (ALS rating scale) (r=0.001, P=0.998). ALS score was negatively correlated with course of disease(r=-0.540, P=0.001).Conclusions Chinese KD pa? tients share similar clinical phenotypes with those of other races but exhibit slightly different clinical characteristics. The length of the CAG repeat influences age at onset but not the severity of disease. Severity of disease is related to the course of disease.

Objective To describe the clinical features, differential diagnosis and therapeutic method of Madras motor neuron disease (MMND) to improve the understanding of MMND. Methods We retrospectively summarized the clinical data of 3 MMND patients. and conducted the related literature review to compare the similarities and differences on clinical features between our cases and foreign MMND patients. Results Patients in the present study were adult-on?set without definite family history. The main manifestations were multiple lower cranial nerve palsies along with weakness and wasting of proximal limbs. Bifacial palsy and dysarthria were most presented in patients, while definite hearing im?pairment was rarely seen. Two patients had fasciculation and atrophy in tongue and one presented with dysphagia. Weak?ness and atrophy were more frequently presented in upper extremities than in lower limbs. All patients had signs of upper motor neuron damage. The level of creatine kinase (CK) moderately increased in one case. Electromyography (EMG) de?tected a widespread neuronal damage in all patients. MMND should be differentiated from Amyotrophic Lateral Sclerosis, Kennedy Disease and Brown–Vialetto–van Laere Syndrome. Intravenous immunoglobulin therapy showed effective in some cases to some extent. Compare to foreign MMND patients, bifacial weakness at onset was more frequently presented in our patients, but hearing impairment was absent. Conclusion The clinical features of MMND include weakness and at?rophy of limbs, involvement of facial and bulbar muscles, pyramidal dysfunction and hearing impairment. Some clinical manifestations of our patients are different from foreign MMND patient.

Objective:To investigate the death of patients with granulocytopenia complicated with infection after chemotherapy and the changes of vital signs before emergency treatment, and to analyze the prognosis of different vital signs on patients' prognosis and emergency treatment.Methods:This study used a case-control study method to select 211 patients with hematologic malignancies who met the inclusion criteria in two tertiary hospitals in Weifang City. The vital signs of patients were collected and the vital signs were analyzed using SPSS 17.0 software. And the statistical significance and predictive value in the emergency response group.Results:The heart rate, respiratory rate, systolic blood pressure, blood oxygen saturation and urine volume were significantly different between the survival group (112 cases) and the death group (99 cases)( t values were 11.038-177.102, P<0.01). The area under the receiver operating characteristic curve of body temperature, heart rate, respiratory rate, systolic blood pressure, and blood oxygenation saturation and urine volume were 0.547, 0.495, 0.294, 0.899, 0.988, and 0.827, respectively. The highest predictive efficiency (higher level) was observed with the change of blood oxygen saturation, and the corresponding optimal cutoff point. 0.91; between the emergency treatment group (103 cases) and the non-emergency treatment group(108 cases), the difference in heart rate, respiratory rate and oxygen saturation between the two groups was statistically significant ( t values were 5.247, 8.001, 9.066, P<0.01). The area under the receiver operating characteristic curve of body temperature, heart rate, respiratory rate, systolic blood pressure, blood oxygen saturation and urine volume were 0.581, 0.732, 0.813, 0.346, 0.102, and 0.543, respectively. Among them, the predicted value of respiratory frequency change was the highest (medium level), which was the best corresponding. The cutoff point was 27.5. Conclusions:Patients with granulocytic infection after malignant hematologic disease will have abnormal changes in vital signs before death and emergency treatment. However, different vital signs have different effects on predicting disease changes, and should focus on respiratory rate and oxygen saturation. Changes, when the respiratory rate exceeds 27 beats/min, the probability that the patient needs to implement emergency treatment such as rescue will increase. If the condition is not effectively controlled, the blood oxygen saturation is lower than 0.91, the risk of death of the patient is greatly increased.

Objective To analyze relation of comprehensive parameters of the dose-volume V5,V10 and V20 with radiation pneumonitis (RP) in patients with advanced stage non-small-cell lung cancer (NSCLC) treated with three-dimensional conformal (3DCRT) or intensity-modulated radiation therapy (IMRT).Methods Data of 90 patients with histologically proved NSCLC treated with 3DCRT or IMRT between November 2006 and July 2009 were collected.The median radiation dose of 70 Gy (range, 61 - 80 Gy) was delivered with late-course accelerated hyperfractionated radiotherapy (LAHRT).The V5 ,V10 ,V20,V30 and mean lung dose (MLD) were calculated from the dose-volume histogram system.The RP was evaluated according to the common toxicity criteria 3.0(CTC 3.0).Results The range of V5 ,V10 and V20 was 36% - 98%, 27% - 78% and 19% - 54%, respectively, with a median value of 66%, 48% and 31%, respectively.The RP of grade 1,2,3,4 and 5 was observed in 29,23,5,1 and 1 patients.The V5,V10 ,V20, contralateral V10, GTV,PTV, and numbers of fields were all significantly associated with RP of ≥grade 1 (χ2=2.04, 2.05, 2.01, 4.62, 6.50, 5.61, 5.61, and P= 0.044, 0.043, 0.047, 0.030,0.010,0.020,0.020).The V5, V10, V20, V30, and MLD were all significantly associated with RP of ≥ grade 2 (χ2= 2.05,2.20,2.96,4.96,5.20, and P = 0.040,0.030,0.000,0.030,0.020).In Logistic regression analysis, GTV was the only factor significantly associated with RP of ≥ grade 1 (χ2= 4.06, P =0.044).The V20 was the only factor significantly associated with RP of ≥grade 2(χ2=9.61,P=0.002).The RP of ≥grade 2 was significantly increased when V20 was more than 31%.The RP of ≥grade 2 was significantly increased when V20, V10 and V5 were more than 31%, 48% and 66%, respectively.The RP of ≥ grade 2 was significantly increased when V20 was more than 31% and V5 was more than 66%.Conclusions The comprehensive parameters combined with V5, V10 and V20 are effective in predicting RP.

Objective:To investigate the clinical effect of minimally invasive catheterization based on computer 3D-Slicer software system in the treatment of hypertensive intracerebral hemorrhage (HICH).Methods:Three hundred and fifty patients with HICH treated in People′s Hospital of Lanling County in Shandongfrom June 2019 to June 2020 were selected as the research object. According to the operation method, they were divided into 3D-Slicer group (175 cases) and CT group (175 cases). They were treated with 3D-Slicer software-assisted minimally invasive catheterization and minimally invasive soft-channel drainage under CT localization, respectively. The general conditions of the surgery, hematoma clearance rate and laboratory indexes, oxidative stress index and prognosis were compared between the two groups.Results:The intraoperative blood loss, the hospitalizationtimein the 3D-Slicer group were lower than those in the CT group: (81.42 ± 12.33) ml vs. (101.54 ± 11.71) ml, (15.67 ± 3.71) d vs. (17.22 ± 3.52) d; the success rate of one-time successful puncture to preset position in the 3D-Slicer group was higher than that in the CT group: 100.00%(175/175) vs. 81.14%(142/175), there were statistical differences ( χ 2 = 34.26, P<0.05). The hematoma clearance rate after the surgery for 1, 3 and 7d in the 3D-Slicer group were higher than those in the CT group:(87.93 ± 8.54)% vs. (66.43 ± 7.99)%, (92.48 ± 10.31)% vs. (89.52 ± 11.74)%, (96.37 ± 10.22)% vs. (94.30 ± 9.25)%, there were statistical differences( P<0.05). After the surgery for 7 d, the levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) in the 3D-Slicer group were higher than those in the CT group: (121.36 ± 10.59)U/L vs. (109.14 ± 9.05) U/L, (92.80 ± 8.63) μg/L vs. (81.45 ± 9.11) μg/L, (24.64 ± 5.43) U/L vs. (20.84 ± 3.47) U/L; while the level of malondialdehyde (MDA) was lower than that in the CT group: (4.42 ± 0.57)μmol/L vs. (5.19 ± 0.51) μmol/L, there were statistical differences ( P<0.05). After the surgery for 3 months, the rate of favorable prognosis in the 3D-Slicer group was higher than that in the CT group 73.71%(129/175) vs. 62.29%(199/175), there was statistical difference ( χ2 = 5.25, P<0.05). Conclusions:Minimally invasive catheterization based on 3D-Slicer software system in the treatment of HICH can not only improve the clinical efficacy, but also shorten the hospitalization time, reduce intraoperative blood loss, and improve the prognosis.