ObjectiveTo learn the rpoB mutation difference in rifampicin/rifabutin cross-resistant (RIF/Rfb-R)clinical isolates and in rifampicin-resistant/rifabutin-susceptible (RIF-R/Rbo-S)clinical isolates of Mycobacterium tuberculosis.Methods To sequence the full-length genome of rpoB gene,and analyze the rpoB full-length gene mutation differences in 278 RIF/Rfb-R clinical isolates,40 RIF-R/Rfb-S clinical isolates,30 rifampicin-susceptible/rifabutin-susceptible (RIF-S/Rfb-S) and in 1 reference strain ofH37Rv.ResultsNo mutations of rpoB full-length gene were found in H37Rv reference strain and RIF-S/Rfb-S clinical isolates.In RIF/Rfb-R clinical isolates,531 (70.5％ ) and 526 (20.9％ ) were the most frequent mutation codons.223 (80.2％ ) isolates possesed single mutations as S531L,S531W,H526D,H526Y,H526R,Q513K,Q513P,Q510H,V176F,P287R,Y395C and H404Y.55 (19.8％) isolates had multiple mutations,and among these the S531L,H526 R,H526Y,H526D,D516G and Q513K were the main substitutions which were in combination with other points.In RIF-R/Rfb-S clinical isolates,516 (65.0％),526 ( 17.5％ ) and 533 ( 10.0％ ) were the most frequent mutation codons.21 (52.5％ ) isolates possesed single mutations as L533P,H526L,H526S,S522L,D516V,D516Y and D516F.19 (47.5％)isolates had multiple mutations and among these the D516V and L533P were the main substitutions which were in combination with other points.CondusionsIn our study,100％ rifamycin-resistant clinical isolates of Mycobacterium tuberculosis had rpoB mutations,but the mutations in RIF/Rfb-R clinical isolates were sharply different from RIF-R/Rfb-S clinical isolates in mutation positions or amino acids substitutions of single mutations strains,mutation positions or combination types and the most frequently mutation codons or amino acids substitutions of multiple mutations strains.Thus,DNA sequencing is instructive and meaningful to choose rifampicin or rifabutin for tuberculosis treatment.

Objective To investigate the expression and significance of caveolin-1 in breast carcinoma.Methods Using immunohistochemical method the protein expression of caveolin-1 were analyzed in 105 cases of breast carcinoma and 50 cases of non-cancerous breast tissues.The relationship between caveolin-1 expression and CK5/6,EGFR,and E-cadherin expression was investigated.Clinical data of 105 cases of breast carcinoma were retrospectively analyzed.Results Among 105 cases of breast carcinoma,there were 20 cases of basal-like subtype,22 cases of luminal subtype A,23 cases of luminal subtype B,23 cases of HER2 over-expressing subtype,17 cases of normal breast-like subtype.Positive rate of caveolin-1 was significantly lower in breast carcinoma than in non-cancerous breast tissues (24.8％ vs.88.0％,P ＜ 0.05).Positive rate of caveolin-1 (75.0％) was higher in breast carcinoma than in luminal subtype A ( 4.8％ ),luminal subtype B ( 17.4％ ),HER2 over-expressing subtype ( 17.4％ ) or normal breast-like subtype( 11.8％ ),all P ＜0.05.Caveolin-1 expression was associated with expression of CK5/6 and EGFR(P ＜0.01 ).In univariate analysis,positive caveolin-1 was associated with higher lymph node metastasis rate (18/26,69.2％ )than negative (37/79,46.8％ ),P =0.047,and shorter 5-year-disease-free survival (38.46％ vs.74.68％,P =0.0004 ),but in multivariate analysis caveolin-1 was not an independent predictor of 5-year- disease-free survival (P ＞ 0.05).Conclusions Caveolin-1 can be seen as a screening mark of basal-like breast carcinoma,it may promote the invasiveness of breast cancer cells,but it is not an independent prognostic predictor of breast cancer patients.

Objective To investigate the role of TNSALP gene detection in prenatal diagnosis of HPP. Method The clinical data and the results of complete exon sequencing of TNSALP gene in one neonate with low alkaline phosphatase (HPP) were analyzed retrospectively. Peripheral bloods from his family members were collected. The amniotic fluid cell in fetuses at 17 weeks was tested for candidate gene mutations by Sanger sequencing. Results Mainly manifestations in 6-day-old baby were multiple fractures, limb shortening and bending and dyspnea. He died of respiratory failure 9 days after birth. The serum alkaline phosphatase was decreased and serum calcium was decreased slightly; serum phosphorus, serum 25 hydroxyvitamin-D and parathyroid hormone were normal. X-ray showed that the whole body bone was very poorly mineralized, and the long diaphysis was enlarged with shape of a cup at the end and multiple fractures existed. Gene sequencing revealed a complex heterozygous missense mutation in the TNSALP gene, including the heterozygous missense mutation c.542C>T in exon sixth causing 181st amino acids changed from serine to leucine (p.S181L), and tenth exon heterozygous missense mutation in c.1016G>A causing 339th amino acid changed from glycine to glutamic acid (p.G339E). The parental phenotypes were normal. The c.542C>T mutation is inherited from his father and the c.1016G>A mutation is inherited from his mother. These two mutations were not detected in the fetus. Conclusion TNSALP gene analysis can be applied to the diagnosis and prenatal diagnosis of HPP.

Objective To explore the feasibility of mechanical chest compression to establish a rat model of car?diopulmonary resuscitation ( CPR) . Methods 4?month old healthy male Sprague Dawley rats were randomly divided into control group ( n=6 ) and model group ( n=10 ) . After induction of anaesthesia with 10% chloraldurate ( 3 ml/kg, i. p. ) , tracheal intubation and left femoral artery cannulation were performed. Under electrocardiographic and artery blood pressure monitoring, tracheal obstruction ( TO) was performed to rats in model group. At 2 min after the cardiac arrest ( CA) occurred, CPRs were administered to the rats using a self?made animal chest compressor, which provided chest?com?pression at a rate of 200 bpm. Results Shortly after TO, rats in the model group had respiratory arrest, cyanosis and ar?rhythmia. Electrocardiography indicated that CA occurred within 4-5 min, with a decreased artery systolic blood pressure ( <40 mmHg) and a zero pulse pressure. Return of spontaneous circulation ( ROSC) after the CPR was successfully a?chieved in 8 rats (80%), with a transient reperfusion arrhythmia. Finally, 60% of the rats (n=6) recovered to con?sciousness and survived for 24 hrs. The serum biochemical analysis indicated that there were electrolyte disturbances, aci?dosis, impaired renal functions and increased myocardial enzyme spectrum. Pathological examination revealed cardiac rhab? domyolysis, no?reflow phenomenon in renal glomeruli, decrease of neurons and pulmonary congestion in the model group rats. Conclusions Mechanical chest compression can provide minimal cardiac output for the requirement of CPR incardiac arrestin rats. It is feasible to establish rat CPR model with the mechanical chest compression.

Objective To perform the bibliometric research on the literatures of the endoscopic retrograde cholangio pancreatog-raphy(ERCP) in the last ten years to provide reference for accurately grasping the international research situation and selecting the cutting-edge technological topics .Methods The literatures of ERCP in the database Web of Science from 2001 to 2011 were collect-ed .The CiteSpace software was adopted to analyze the distribution characteristics ,key point literature ,hotspots fields and research frontier of articles of ERCP .Results 2 921 citations of the research articles were retrieved .The analysis showed that America was strongest in the ERCP research field ;10 key point articles were detected .The high frequency keywords were complications ,manage-ment ,sphincterotomy ,etc .,the concentricity ranged 0 .06-0 .24 .The high frequency burst words were gold standard ,procedure-re-lated-complications ,biliary-stents ,but the concentricity was 0 .Conclusion The research hotspots of ERCP field are the occurrence rate of ERCP postoperative complications ,diagnosis and treatment ,but no obvious research cutting-edge is formed .

Objective To study the effect of neuronal Nogo-66 receptor (NgR1) antagonist,soluble Nogo-66 receptor (sNgR1-Fc),on promoting the endogenous neural precursor cells (NPCs) differentiating into neurons in order to clarify the mechanism.Methods The cortical infarction was induced by photochemistry,named photothrombotic cortical injury (PCI).Twelve Sprague Dawley rats were randomly divided (random number) into three groups:Sham-operated group,PBS group,and sNgR1-Fc group.PBS (PBS group) or sNgR1-Fc (sNgR1-Fc group) was injected into the lateral ventricle of brain with a minipump.BrdU (Bromodeoxyuridine) was injected into the peritoneal cavity 4-6 days after PCI.The subdentate gyrus zone (SGZ) of brain from sacrificed rat was harvested for Immunohistochemistry to observe the ratio of NeuN +/BrdU + cells 35 days after PCI.Proteins including Nestin、Notch1 and Mash1 were detected by Western Blot.Results The cortical infarction in rat was successfully induced by photochemistry.Thirty-five days after PCI,the BrdU + cells number and theratio of NeuN +/BrdU + in the SGZ of the ipsilateral cerebrum hemisphere with PCI were significantly higher in sNgR1-Fc group than those in PBS group (P ＜ 0.05).The levels of Notch1,Mash1 and Neuro D in the sNgR1-Fc group were significantly higher than those in the PBS group (P ＜ 0.05),which were significantly higher than those in the Sham-operated group.Conclusions sNgR1-Fc could promote the endogenous NPCs differentiating into neurons in a cortical infarction model.The mechanisms may be attributed to the Notch/bHLH (proneural basic helix-loop-helix genes) signaling way.

Objective To investigate the influence of low-dose,long-term arsenite exposure on keratinocyte (HaCat) cell apoptosis and the expression of Bcl-2 and Bax proteins.Methods HaCat cells were exposed to different concentrations of NaAsO2[0(control group),0.05,0.10 μmol/L] for 15 weeks.A total of 10 000 cells in each group were measured to detect the level of apoptosis by flow cytometry.The protein expressions of Bcl-2 and Bax were determined by Western blotting method.Results The cell apoptosis rate was significantly different between groups (F =17.19,P ＜ 0.05).Compared with that of the control [(1.42 ± 0.13)％],the apoptosis rates of the 0.05 and 0.10 μmol/L groups [(1.23 ± 0.08)％ and (1.04 ± 0.06)％] decreased significantly (all P ＜ 0.05); the 0.10 μmol/L group was significantly lower than that of the 0.05 μmol/L group(P ＜ 0.05).The protein expressions of Bcl-2 and Bax were significantly different between groups (F=107.38,346.45,all P＜ 0.05).The protein expression of Bcl-2 of the 0.05 and 0.10 μmol/L groups were (143.89 ± 7.74)％ and (199.96 ± 12.18)％,respectively,which were significantly higher than that of the control group[(100.00 ± 1.45)％,all P ＜ 0.05] ; the 0.10 μmol/L group was significantly higher than that of the 0.05 μmol/L group (P ＜ 0.05).The protein expression of Bax of the 0.05 and 0.10 μmol/L groups were (70.78 ± 1.53)％ and (54.80 ± 1.34)％,respectively,which were markedly lower than that of the control group[(100.00 ± 3.09)％,all P ＜ 0.05]; the 0.10 μmoL/L group was significantly lower than that of the 0.05 μmol/L group(P ＜ 0.05).Conclusion Decreased level of apoptosis induced by low level and long-term arsenic exposure may be associated with increased expression of Bcl-2 protein and decreased expression of Bax protein.

Objective To investigate the feasibility of iterative model reconstruction (IMR) combined with low tube voltage and low iodine intaken of contrast media in head and neck CT angiography (CTA). Methods Eighty patients with clinical suspicion of head and neck vascular disease underwent head and neck CTA were enrolled in this prospective study. According to random number table, patients were randomly divided into two groups. Group A (n=40) was scanned according to the protocol of 120 kV, 50 ml iopromide (370 mg/ml) and filtered back projection (FBP) reconstruction, and group B (n=40) was scanned with 80 kV, 30 ml iohexol (300 mg/ml) and IMR, while the other parameters kept consistent. Mann-Whitney U test was used to compare the enhanced CT value of arteries, image noise, signal noise ratio (SNR), contrast noise ratio (CNR), the score of image quality, effective radiation dose (ED) and iodine intaken of contrast media between two groups. Results The CT value at the origin level of common carotid artery were (316.9 ± 53.0) and (433.4 ± 101.8)HU in group A and B, image noise were (28.1 ± 6.8)and (12.1 ± 2.6)HU ,SNR were (11.9 ± 3.2) and (37.7 ± 13.3) ,and CNR were (10.2 ± 2.9) and (32.6 ± 13.3), respectively. There were showed significant differences in CT value, image noise, SNR and CNR between two groups (Z=-5.490,-7.592,-7.698,-7.660, P<0.05). The CT value at the origin level of internal carotid artery were (359.5 ± 54.3) and (443.5 ± 120.1) HU in group A and B, image noise were (18.8±6.2) and (6.8±1.7) HU ,SNR were 21.5±8.7 and 69.7±27.4 ,and CNR were 18.0±7.3 and 62.7± 26.4, respectively. The significant differences were acquired in CT value, image noise, SNR and CNR between two groups (Z=-3.022,-7.376,-7.496,-7.515, P<0.05). The CT value at M1 segment level of middle cerebral artery were (321.1±47.3) and (401.6±104.0) HU in group A and B, image noise were (32.3± 17.2) and (11.2 ± 2.7) HU,SNR were 12.4 ± 5.6 and 39.3 ± 18.4,and CNR were 10.7 ± 4.7 and 36.4 ± 17.7, respectively. There also showed significant differences in CT value, image noise, SNR and CNR between two groups (Z=-3.527,-7.487,-7.482,-7.535, P<0.05). The score of image quality of group A and B were 3.9 ± 0.7 and 4.5±0.6, which also showed significant difference between two two groups (Z=-3.517, P<0.05). The ED were (2.78 ± 0.13) and (0.84 ± 0.04) mSv for group A and B, which also showed significant difference between the two groups (Z=-7.706, P<0.05). The iodine intaken of contrast media were 1.85 g and 0.90 g in group A and B. Conclusion IMR combined with low tube voltage and low iodine intaken of contrast media can not only decrease effective radiation dose and iodine intaken of contrast media significantly, but also improve the imaging quality in head and neck CTA examination.

Objective To explore the impact of four different anesthetic drugs commonly used in animal experi-ments on cardiovascular system in rats.Methods Electrocardiogram ( ECG) and blood pressure were dynamically recor-ded by a BioPac MP150 system after anesthesia.In addition, the blood glucose at different time points and hepatic func-tion, kidney function, cardiac enzymes and electrolytes at the end of the test were collected.Rusults Chloral hydrate caused severe ventricular arrhythmia.Isoflurane had inhibitory effect on the heart rate.Pentobarbital sodium induced a in-crease of ECG P wave.Urethane caused J point elevation of ECG.Blood pressure in the urethane-and pentobarbital sodi-um-treated groups were increased.Chloral hydrate caused CK to be raised, while isoflurane showed the opposite effect on CK and CKMB.Alanine aminotransferase and aspartate aminotransferase in the pentobarbitol sodium and isoflurane groups were decreased.Creatinine in the chloral hydrate, pentobarbital sodium and isoflurane groups were lower, and the serum sodium and potassium were decreased in the four groups.Conclusions Chloral hydrate has obvious effect on the cardio-vascular system, and is not suitable for animal studies on cardiovascular diseases.Pentobarbital sodium, urethane, isoflu-rane can be chosen for animal studies on cardiovascular diseases.

Adenine ; analogs & derivatives ; Fanconi Syndrome ; Humans ; Organophosphonates