1.Construction and application of N-terminal Strep-tagged protein expression vector
Yu SHI ; Hongzhang JI ; Xiaofeng BAO
Chinese Pharmacological Bulletin 2016;(1):98-102
Aims To construct the N-terminal Strep-tagged ( NS-tagged) fusion protein expression vector, and to apply the vector to express NS-tagged fusion proteins of Chlamydia RNA polymerase subunit. Meth-ods By using PCR method, NS fusion protein tag and a new multiple cloning sites (MCS) were inserted into pET21c-DH plasmid by primers to replace the original T7 protein tag and MCS. The newly introduced Not I cutting site was chosen for self-ligation of PCR prod-uct. Then, the cyclized PCR product was transformed into DH-5α competent cells. The positive clones were selected by PCR and sequencing. To get NS-tagged fu-sion proteins of chlamydial RNA polymerase subunits, the α, β and β′ subunits were inserted between BamH I and Sal I cutting sites of the newly constructed ex-pression vector. Then, the NS-α, NS-β and NS-β′ ex-pression vectors were transformed into Arctic Express expression cells. The fusion protein expression statuses of transformed cells were identified by Commassie blue staining and Western blot. Results The NS-tagged fusion protein expression vector pET21c-NS-MCS was successfully constructed, and NS-α, NS-β and NS-β′fusion proteins were obtained by using this newly con-structed expression vector. Conclusions In this pro-ject, we constructed an NS-tagged fusion protein ex-pression vector and applied it to express NS-α, NS-βand NS-β′ fusion proteins. Our study can lay a solid foundation for the study of transcriptional regulation of Chlamydia genes.
2.The research and strategy about the current situation about the analysis of cellular morphology in Tibet region
Quangui SHI ; Dongfang FENG ; Sijuan DING ; Hongzhang CHEN ; Baohong QI ; Huaying SHI
International Journal of Laboratory Medicine 2014;(8):1007-1008
Objective To understand the current situation and level of clinical laboratory about the analysis of cellular morphology in Tibet region .Methods Authors investigated the information about the staff of clinical laboratory testing the patient′s blood smear under microscope ,executing the rules and regulations by using standard questionnaires .Results Some of the clinical laboratory didn′t founded the rule and standardization of rechecking about abnormal blood routine (5/15 ,33 .3% ) .Some of the divi-sion leadership didn′t pay enough attention to the staff′s basic operation (2/15 ,13 .3% ) .Most of the staff didn′t being trained about cellular morphology in special purpose workshop (6/43 ,88 .8% ) .Some of the hospital didn′t carried out the chemistry stai-ning about blood cells(10/15 ,66 .7% ) .Conclusion It′s important to promote the quality and level about the analysis of cellular morphology in Tibet region .
3.Endovascular embolization for the management of cryptogenic massive hemoptysis in long-term smokers
Sen JIANG ; Hongzhang SHI ; Xiwen SUN ; Ping JIANG ; Li ZHANG ; Bing JIE ; Dong YU
Chinese Journal of Radiology 2011;45(12):1199-1202
ObjectiveTo determine the effectiveness of endovascular embolization in patients with cryptogenic massive hemoptysis who were all long-term smokers.Methods Aortography and subclavian artery angiography were performed in 21long-term smokers with cryptogenicmassive hemoptysis.Transarterial embolization (TAE) was performed in patients with detectable pathologic systemic arteries.The angiographic findings were reviewed and the clinical and follow-up CT results were observed.ResultsThe pathologic systemic arteries were all bronchial arteries (BAs) and thirty-five arteries were involved.The angiography demonstrated peripheral hyperplasia in all BAs,with 24 pathologic BAs supplying the right lung and 25 supplying the upper lobes.In thirty-five BAs,24 showed hypertrophy and 11 were normal.TAE of the pathologic BAs was successfully performed and cessation of bleeding was achieved in all patients.During follow-up,one patient had episodic bloody sputum after embolization and no recurrence in all patients.The follow-up CTdemonstratednoadditionalabnormalitybesidespre-existingpulmonaryemphysema.Conclusion Cryptogenic massive hemoptysis in long-term smokers efficiently treated by endovascular embolization of the responsible bronchial artery.
4.Effects of prophylactic antibiotics administration on colonized microorganisms of thoracic catheter in pneumothorax patients treated with closed thoracic drainage
Li ZHANG ; Jinming LIU ; Hongzhang SHI ; Ping JIANG ; Hongbin JIANG ; Jian MA ; Yu CHEN
Chinese Journal of Emergency Medicine 2011;20(3):287-291
Objective To investigate the feature of the microorganisms colonization of the thoracic catheter-related infection and evaluate the clinical significance of prophylactic antibiotics administration in patients with pneumothorax treated with closed thoracic drainage. Method A total of 120 patients with pneumothorax treated with closed thoracic dramage in emergency department wore enrolled. The patients were randomized (random number) into group A (n =60) and group B (n =60). In group A, the patients received levofloxacin mesylate injection and in group B, patients received physiological saline injection instead after closed thoracic drainage. The tip of catheter was cut off to get a 2-cm long segment after catheter removal and this segment was dipped into a bottle filled with liquid culture medium for microorganism culture. Statistical analysis carried out by using χ2 test or Fisher exact test. Results Of all 120 patients, microorganisms were found in 49 segments of catheter and 57 strains of microorganisms were found. The four most common microorganisms were Coagulase-negative staphylococci (57.9%), Candida albicans (10. 5%),Staphylococcus aureus (7%) and Acinetobacter baumanii (7%). All of them were highly drug-resistant to β-1actam antibiotics. The difference in the positive rate of microorganism culture was distinct in pneumothorax patients with underlying diseases (50%) in comparison to the patients without underlying diseases (31%) (P < 0.05). The positive rate of microorganism culture increased significantly as the duration of drainage was longer than 14 days (P < 0.01). The positive rate of culture in group A was lower than that in group B if the duration of drainage was less than 7 days (8.3% vs 52.9%, P < 0.01). The positive rate of culture after drainage for 7 days was 21.4% in group A and 68.8% in group B (P <0.05), and that after drainage for over 14 days was 70% in both groups (P > 0.05). There were no significant differences in outcome and days of hospital stay between two groups (P > 0. 05). Conclusions The common colonized microorganisms of thoracic catheter-related infection are conditional pathogens and highly resistant to antibiotics. Lengthening the duration of drainage and having underlying diseases increase the risk of infection. Although prophylactic antibiotics administration is beneficial to decrease the risk of thoracic catheter-related infection, it has no effects on shortening hospital stay and outcome of disease.
5.Diagnostic role of endobronchial ultrasound-guided transbronchial needle aspiration( EBUS-TBNA )in the smear and culture negative pulmonary tuberculosis
Hongbin JIANG ; Hao WANG ; Gening JIANG ; Chunyan WU ; Xiao ZHOU ; Shengxiang REN ; Aiwu LI ; Hongzhang SHI
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(9):526-528
Objective To evaluate the diagnostic role of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the smear and culture negative tuberculosis.Methods The tuberculosis suspected patients with spu-tum-negative for three times and with hilar or mediastinal lymphadenopathy were randomly divided into 2 groups,study group received EBUS-TBNA and bronchoalveolar lavage group(BAL) examination,control group received BAL examination only.Chi-square test was used to compare their diagnostic sensitivity and negative predictive value.Results Totally 82 patients were included this study,40 patients were in control group and 42 in the study group.In the control group,tuberculosis was confumed in only 8 cases by means of bacteriological examination in the BAL fluid,27 were confirmed by the following surgery ordiagnostic treatment of anti-Tuberculosis,and the other 5 cases were diagnosis as other diseases; In the study group,tuberculosis was confirmed in 28 patients through bacteriological and pathological examination,8 were false negative and the other 6 were diagnosis as non-tuberculosis diseases.The diagnostic sensitivity in the study group was significantly higher than that in control group (77.8% verus 22.9%,x2 =21.4,P < 0.01 ) ; and the negative predictive value in the EBUS-TBNA group was also significantly higher than that in the BAL group (42.9% VS 15.6%,x2 =3.97,P =0.046).Complications were similar inthese 2 groups,only 1 case of intervention required puncture site bleeding happened in the study group.Conclusion EBUSTBNA has a higher sensitivity and negative predictive value in the diagnosis of smear and culture negative pulmorary tuberculosis patients with hilar or mediastinal lymph nodes.This technique is a safe method with few complications than the traditional BAL examination.It may play an important role in the diagnosis of smear and culture negative tuberculosis patients.
6.Influencing factors for radial shortening after surgical operation for intra-articular fractures of the distal radius
Haolin SUN ; Chunde LI ; Xianyi LIU ; Hong LI ; Yongping CAO ; Weibing CHAI ; Hong LIU ; Xuedong SHI ; Hongzhang LU
Chinese Journal of Trauma 2011;27(8):694-697
ObjectiveTo analyze the influencing factors and clinical results of radial shortening after operation of intra-articular fractures. MethodsA total of 54 patients with intra-articular fractures of the distal radius treated surgically from January 2003 to January 2008 were followed up and divided into radial shortening group ( > 4 mm) and control group ( < 4 mm) by whether radial shortening was more than 4 mm. Fracture types, comminution and defection degree, age, surgical approach and internal fixation method, whether bone implant in operation, post-operative volar tilting angle and ulnar inclination angle, and wrist function score were investigated and compared between the two groups. ResultsThe incidence of radial shortening in post-operation of intra-articular fractures of the distal radius was 30%.Age and whether bone implant in operation were significantly different between the two groups. Type C fractures were more liable to radial shortening than type B fractures. Operation approach and internal fixation method have no significant influence on the radial shortening. Volar tilting angle and ulnar inclination angle were not significantly different between the two groups and the excellent and good rate of function score was lower in the radial shortening group. ConclusionThere is a high incidence of radial shortening after operation for the intra-articular fractures of the distal radius, which affects the clinical results of operation. Therefore, we should pay more attention to the influencing factors and take corresponding precaution methods during operation.