Brucella organisms are facultative intracellular bacteria capable of surviving inside professional and non-professional phagocytes.Upon cell contact the bacteria is internalized via receptor molecules.Once inside cells,Brucella localizes in early phagosomes,where it avoids fusion with late endosomes and lysosomes.Then,the bacterium redirects its trafficking to autophagosomes and finally reaches the endoplasmic reticulum,the replicating niche.Once inside the endoplasmic reticulum,Brucella extensively replicates without restricting basic cellular functions or inducing damage to cells.Invasion,intracellular trafficking and replication of Brucella organisms in professional and non-professional phagocytes and the molecular determinants involving Brucella intracellular life are reviewed in this article.

Objective: Screening the influencing factors that affect the indigo-generated significantly in processing of Indigo Naturalis, the levels were optimized and combined to obtain the optimum technology for the indigo-generated. Methods: Using an HPLC method for determining indigo in crude Indigo Naturalis and taking indigo content generated from fresh leaf as an index, the significantly influencing factors and the optimal level combination were screened by the statistical methods of Plackett-Burman design and Box-Behnken design response surface analysis. Results: The soaking and making indigo process of Indigo Naturalis was identified as the wax was not removed before soaking, soaking pH value was 3.5, and solvent volume (mL) : leaf mass (g) was 13. Soaked in dark at 25 °C for 24 h, and ventilation time of 30 min, then ammonia water was added to adjust pH value to 9. Conclusion: The soaking and making indigo process of Indigo Naturalis could be optimized systematically, it could be used to clarify the various factors on the impact of the active indigo in the process of Indigo Naturalis. At the same time, it could lay the foundation for the study on processing principle of Indigo Naturalis.

Objective To evaluate the ability of further discriminating diagnosis of the headchest lead electrocardiogram (HCECG) in elderly patients with abnormal Q waves in routine lead electrocardiogram (RLECG) in inferior lead. Methods The 55 male patients, aged 65-88 years,with abnormal Q waves in both lead Ⅲ and aVF were selected and divided into two groups: myocardial infarction (MI) group and non-MI group, according to if the patient had a history of acute MI. All the patients accepted examination of coronary computed tomographic angiography (CTA) and ultrasound cardiogram, those with Wolff-Parkinson-White syndrome and myocardial hypertrophy were excluded.The 30 individuals of control group had no abnormal Q waves in lead Ⅱ , Ⅲ and aVF. HCECG and RLECG were recorded simultaneously in respective groups and occurrence rate of Q waves in correlative leads Ⅱ, Ⅲ, aVF and HL3, H0, HR3 were matched and compared, sensitivity and specificity were compared in respective leads. Results There were serious stenosis in 22 patients (100%) in MI group, and there were mild stenosis in 10 (30. 3%) and moderate stenosis in 23 patients (69.7%) in non-MI group. There was no significant difference between HCECG and RLECG in occurrence rate of Q waves in MI group (P＞ 0. 05 ). Non-MI group left anterior axillary line, Ⅱ ,Ⅲ, right anterior axillary line, near the anterior midline, aVF without Q wave and exclusion of old MI diagnostic specificity were 100%, 97.0%(32/33), 97.0% (32/33), 15.2% (5/33), 100%, 39.4%( 13/33)respectively. Conclusions Pseudo-changes are rarely found in HCECG and there is a higher degree of conformity in HCECG with coronary lesions, therefore HCECG may be used to discriminate whether the inferior abnormal Q waves occurred in RLECG are truly abnormal or not.

ObjectiveTo study the expression changes of peripheral blood monocyte human leukocyte antigen DR (HLA-DR) in patients with severe craniocerebral injury,and investigate the correlation between HLA-DR expression and infection and prognosis.MethodsNinety patients with craniocerebral injury were selected as experimental group and were divided according to the Glasgow coma scale (GCS) score after hospitalization into experimental group 1 (GCS score 13-15 scores ),experimental group 2 (GCS score 9-12 scores) and experimental group 3 (GCS score 3-8 scores) with 30 patients each,which were moderate,medium,severe craniocerebral injury,respectively.Thirty healthy people were chosen at the same period as control group.The HLA-DR expression of experimental group was detected after 1,3,7 and 14 d of admission by flow cytometry,and the HLA-DR expression of control group was detected on the day they got physical examination.The rates of infection,cure,disability,vegetative state and mortality were counted after 30 d of admission.ResultsThe HLA-DR expressions in experimental group 1 and experimental group 2 after 1,3,7,14 d of admission were (28.11 ± 2.37),(26.45 ± 1.63),(27.75 ± 1.83),(27.15 ± 2.17) MCF and (29.34 ±2.07),(27.55 ± 1.63),(28.42 ± 1.94),(29.46 ±2.12) MCF,which had no statistical difference compared with that in control group [(29.18 ± 1.91 ) MCF](P＞ 0.05).The HLA-DR expressions in experimental group 1 and experimental group 2 after 1,3,7 d of admission and control group had statistical differences compared with those in experimental group 3 after 1,3,7 d of admission [(18.02 ± 1.78),(16.05 ± 1.97 ),(20.76 ± 1.65) MCF ] (P ＜ 0.05).The HLA-DR expressions in experimental group 1 and experimental group 2 after 14 d of admission and control group had no statistical significance compared with that in experimental group 3 after 14 d of admission [ (26.13 ± 2.15) MCF](P＞ 0.05).The infection rates of experimental group 1,experimental group 2 and experimental group 3 were 0,3.6％(1/28),82.8％(24/29),respectively,while the cure rates were 100.0％ (30/30),100.0％ (28/28),10.3％ (3/29),the disability rates were 0,0,41.4％ (12/29),the vegetative state rates were 0,0,20.7％ (6/29),and the mortality were 0,0,27.6％ (8/29).There was no statistical significance in the rates of infection,cure,disability,vegetative state and mortality between experimental group 1 and experimental group 2 (P＞ 0.05 ).While there was statistical differences in the rates of infection,cure,disability,vegetative state and mortality among experimental group 1,experimental group 2 and experimental group 3 (P ＜ 0.05).ConclusionsThe HLA-DR expression changes of patients with moderate and medium craniocerebral injury after 1,3,7,14 d of admission are not significant.The HLA-DR expression of patients with severe craniocerebral injury begins to decline from 1 d after injury,declines obviously at 3 d,increases from 7 d,returns to normal level at 14 d.The decline of HLA-DR expression in patients with severe craniocerebral injury is correlated with the infection,and predicts poor prognosis.

Objective To investigate the efficacy and safety of isosorbide mononitrate sustained release tables plus vinorelbine and cisplatin in patients with previously untreated advanced non-small-cell lung cancer (NSCLC).Methods One hundred and ten patients with stage ⅢB-Ⅳ NSCLC were randomly assigned to group A (57 cases) and group B (53 cases) by random mumber table.Patients in group A were treated with vinorelbine 25 mg/m2 on the first and eighth day and cisplatin 25 mg/m2 on day 2-4,with transdermally applied isosorbide mononitrate sustained release tables (40 mg,daily for 8 days),and patients in group B were treated with vinorelbine and cisplatin.Response to treatment was assessed by RECIST1.1 and adverse effect was assessed by NCI-CTC(3.0).Results The response rate in group A (58.2％,32/55 patients) was significandy higher than that for patients in group B (30.8％,16/52 patients; x2 =8.120,P =0.004).Median TTP and median OS in group A were longer than those in group B (8.2 vs 5.8 months,x2 =10.684,P =0.001 ; 11.6 vs 9.0 months,x2 =11.231,P =0.001).While,patients with squamous carcinoma showed better response to chemotherapy (RR =2.438,95％ CI:1.136-5.231,P =0.022).Adverse effect difference was not significant between group A and group B,except headache.The rate of grade 1 to 2 headache in group A (34.5％ ; 19 of 55 patients) was significantly higher than that in group B (3.8％ ; 2 of 52 patients; P ＜0.001).Conclusion Use of isosorbide mononitrate sustained release tables combined with vinorelbine and cisplatin may improve overall response,TTP and OS in patients with advanced stage NSCLC.

0.05).The survival area and capillary density were more favorable in the EPCs-injection sites than the controls(P

Objective To observe the different effects of iodine excess on inducing two strain mice thyroiditis. Methods NOD and Balb/c mice, each having 14 mice, were divided into NaI and control group. The mice were given 0.05% NaI water for 8 weeks in NaI group. RIA and ELISA were used respectively to detect TT4, TgAb, TPOAb and TSH level in serum. Morphology changes of thyroid and apoptosis of thyrocytes stained by immunohistochemistry were observed under light microscope. Lymphocytic proliferation of cervical lymph node and spleen to responding to Tg were detected by MTr method. Results After intake of iodine water for 8 weeks, NOD and Balb/c mice showed relative quality of thyroid in Nal group[(104.83±14.52), (155.79±20.77)mg/kg]obviously increased compared with control group[(71.80±20.42), (105.15±21.98)mg/kg, t values:-3.293,-4.429, all P< 0.01)], enlarged follicular lumen with colloid accumulation were observed in thyroid. Serum level of TT4 in Nal group [(29.52±4.42), (19.53± 2.35)nmol/L]to control group[33.40±5.38), (23.47±6.22)nmol/L]of NOD and Balb/c mice showed a decreasing tendency(t values: 1.374,1.567, all P > 0.05). TSH of Nal group showed an increasing tendency in Balb/c mice[(4.14±1.71)μg/L, compared with control [(3.55±1.41)μg/L, t values:-0.705, P > 0.05]and obviously increased in NOD mice [(6.98±0.66)μg/L, compared with control[(555±056)μg/L, t values:-3.562, P< 0.01], but no change of TgAb and TPOAb level in Nal group(1281,1364 cpm, 2.50×103, 0.14×103U/L were observed, compared with control(1297,1220 cpm, 3.17×103,0.03×103 U/L; Zvalues:-0.081,-0.703, -0.244,-1.293, all P > 0.05). In NOD mice NaI group, apoptosis of thyrocytes was more intense than Balb/c mice, obvious infiltration of lymphoeytes, disorganization and focus fibrosis was seen in thyroid. The cell amount of NaI group increased in NOD mice lymph node and spleen cells[(1.100±0.014), (1.076±0.033)]were more than that in the control group [(0.993±0.011), (1.005±0.003), t value:-11.672,-4.314, P < 0.01). Conclusions Iodine leads to enlargement of thyroid and malfunction of thyroid in Balb/c mice. Besides, NOD mice have generate inflammatory reaction in thyroid and produced sensitized lymphocytes to Tg. Iodine excess can induce NOD mice to occur autoimmune thyroiditis.

Objective: To detect the expression of MMP-13 in the skin of mice treated by near-infrared (IRA) and ultraviolet (UV) so as to observe their effects on skin photoaging and their interrelation and explore molecular mechanisms in IRA-induced skin photoaging. Methods: Male ICR mice were randomly divided into five groups: control group (without ray exposure), IRA group, UV group, IRA/UV group, and UV/IRA group. The mice in the latter four groups had their dorsal skin exposed to different radiated ray respectively. The levels of MMP-13 protein and mRNA in the exposed skin were detected by HE, immunohistochemical method, Western blot and real-time PCR. Results: Both skin lesions by visual inspection and H&E staining results showed that mice in the other four groups had skin photoaging in the exposed skin area compared with the control group. The levels of MMP-13 protein and mRNA in the exposed skin in IRA/UV and UV/IRA groups were significantly higher than those in the control mice (P<0.05). In addition, mice in IRA/UV group showed higher levels of MMP-13 protein and mRNA than those in UV/IRA group (P<0.05). Conclusion: ① IRA causes skin photoaging in mice. ② UV and IRA interact with each other, up-regulate the expression of MMP-13, and promote each other in the process of photoaging. ③ The effects of IRA and UV in combination on skin photoaging are closely related to order of exposure. Taken together, avoiding IRA exposure and the expression of MMP-13 play an important role in preventing skin wrinkle formation and treatment of photoaging in mice.

The total RNA was extracted from ginseng leaves of Panax ginseng. The Cu/Zn-SOD gene was amplified via RT-PCR and the pET-28(a)-Cu/Zn-SOD expression vector was constructed. The pET-28 (a)-Cu/Zn-SOD recombinant plasmid was transformed into Escherichia coli BL21 (DE3) competent cells and was induced by IPTG in order to select optimal induction of expression conditions. The target protein was purified by the nickel ions (Ni ) affinity chromatography and the target protein enzyme activity was determinated by the xanthine oxidase method. The similarity of the Cu/Zn-SOD gene sequences and the Cu/Zn-SOD gene sequences of Korean ginseng in NCBI was 99. 00%. The target protein expression level was about 44.42%, and the molecular weight was 16.30 kDa after the pET-28(a)-Cu/Zn-SOD recombinants were induced by IPTG. The purified Cu/Zn-SOD protease activity reached 10,596.69 U x mg(-1). The P. ginseng pET-28(a)-Cu/Zn-SOD prokaryotic expression vector was built by the method of molecular biology, which provided the foundation for studying the Cu/Zn-SOD biology function.
Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Panax ; enzymology ; genetics ; Sequence Analysis ; Superoxide Dismutase ; genetics ; isolation & purification ; metabolism

To reveal the roles of uncoupling proteins (UCPs) in disorder of mitochondrial oxidative phosphorylation induced by free fatty acid during hypoxic exposure, the effects of palmitic acid on activity of UCPs, proton leak and mitochondrial membrane potential in hypoxia-exposed rat brain mitochondria were observed in vitro. Adult Sprague-Dawley (SD) rats were set randomly into control, acute hypoxia and chronic hypoxia groups (n=8 in each group). The acute and chronic hypoxic rats were exposed to simulated 5000 m high altitude in a hypobaric chamber 23 h/d for 3 d and 30 d, respectively. The brain mitochondria were isolated by centrifugation. UCP content and activity were detected by [(3)H]-GTP binding method. The proton leak was measured by TPMP(+) electrode and oxygen electrode. The membrane potential of mitochondria was calculated by detecting the fluorescence from Rodamine 123. Hypoxic exposure resulted in an increase in UCP activity and content as well as proton leak, but a decrease in the membrane potential of rat brain mitochondria. Palmitic acid resulted in further increases in UCP activity and content as well as proton leak, and further decrease in membrane potential of brain mitochondria in vitro from hypoxia-exposed rats, but hypoxic exposure decreased the reactivity of cerebral mitochondria to palmitic acid, especially in the acute hypoxia group. There was a negative correlation between mitochondrial proton leak and K(d) value (representing derivative of UCP activity, P<0.01, r = -0.906), and a positive correlation between proton leak and B(max) (representing the maximal content of UCPs in mitochondrial inner membrane, P<0.01, r = 0.856). Cerebral mitochondrial membrane potential was negatively correlated with proton leak (P<0.01, r = -0.880). It is suggested that hypoxia-induced proton leak enhancement and membrane potential decrease are correlated with the increased activity of UCPs. Hypoxia can also decrease the sensitivity of cerebral mitochondria to palmitic acid, which may be a self-protective mechanism in high altitude environment.
Altitude ; Animals ; Brain ; metabolism ; Cell Hypoxia ; Ion Channels ; metabolism ; Membrane Potential, Mitochondrial ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Oxidative Phosphorylation ; Oxygen ; metabolism ; Palmitic Acid ; pharmacology ; Protons ; Rats ; Rats, Sprague-Dawley ; Uncoupling Protein 1