Objective To research on reversal mechanism of active ingredients of Sanhuang Xiexin Decoction on lipopolysaccharide-induced acute kidney injury.Methods C57BL/6 mice were randomly divided into 6 groups: control group, LPS group, rhein group, berberine group, baicalin group and dexamethasone group.8 mice in each group.Control group were injected intraperitoneally with 0.9%NaCl 0.5 mL, and the remaining groups were injected intraperitoneally with LPS 0.1 mL/g.The mice of rhein group, berberine group, baicalin group and dexamethasone group were respectively injected intraperitoneally rhein, berberine, baicalin ( 0.1 mL/g ) and dexamethasone ( 0.0001 mL/g ) 2 h before LPS injection.One day after LPS treatment, mice were decapitated and blood collected, diagnostic kits and automatic biochemical analyzer were used to detected urea nitrogen and creatinine.TNF-αand IL-6 concentrations in serum were detected by ELISA kit.HE staining and immunohistochemistry were performed in the kidney tissues.expression of apoptosis protein detected by Western blot.ResuIts Compared to control group, the blood urea nitrogen, serum creatnine, TNF-αand IL-6 level were significantly lower ( P<0.05 ) .HE staining found the renal tubular epithelial cell which treated with LPS were swelling, degeneration, loss, narrow lumen and tube type and exudate, renal interstitial edema, inflammatory cells infiltration.And symptoms of the other group were significantly slight.Immunohistochemical results showed that the expression of Bax and Caspase3 in LPS group was obviously increased, while the expression of the others group was decreased, neither did Bcl-2.Western blot results showed that apoptosis proteins Bax, Cleave-caspase3, CytoC expression increased of LPS group, while the other groups were lower than LPS group.But Bcl-2 expression reduced in LPS group and increased in others group.ConcIusion The effective constituent of Sanhuang Xiexin Decoction can reversal of LPS induced acute kidney injury and its mechanism of action is similar to that of dexamethason.

Objective To investigate the genotypic frequency of glucose-6-phosphate dehydrogenase (G6PD) (1376G ＞ T),G6PD 1311C ＞ T and G6PD IVS11-93T ＞ C in 50 newborns with G6PD deficiency in Guangdong region.Methods To identify G6PD deficiency in the patients of neonatal ward in Guangzhou First People's Hospital during 2010,detected by methemoglobin reduction test and measurement of G6PD/6-phosphate dehydrogenase (6PGD) ratio.Fifty G6PD deficiency subjects were classified into the experimental group,20 neonatal jaundice subjects were classified into the control group.Genomic DNA was extracted by standard method from the peripheral blood of each subject.PCR-direct DNA sequence analysis was used to identify G6PD 1376G ＞ T,1311C ＞ T and 11 intron 93T ＞ C mutations.Results The overall results of mutation analysis in the 50 G6PD deficiency subjects showed the existence of 3 different alleles:G6PD 1376G ＞T,1311C ＞T,11 intron 93T ＞ C(including 1311C ＞ T/IVS11-93T ＞ C and 1376G ＞T/1311C ＞T/IVS11-93T ＞ C complex mutations).The different genotypic frequency in the experimental group was G6PD 1376G ＞T 26.0％ (13/50 cases),1311C ＞ T 4.0％ (2/50 cases),11 intron 93T ＞ C 4.0％ (2/50 cases),1311C ＞ T/IVS11-93T ＞ C 2.0％ (1/50 cases),1376G ＞ T/1311C ＞ T/IVS11-93T ＞ C 2.0％ (1/50 cases).The G6PD enzyme activity of the subjects with 1311C ＞ T/IVS11-93T ＞ C and 1376G ＞ T/1311C ＞ T/IVS11-93T ＞ C complex mutation were reduced.These G6PD mutations were not found in the controls.Conclusions G6PD 1376G ＞ T is one of the commonest mutation in G6PD deficiency newborn in Guangdong region.A novel complex mutation 1376G ＞ T/1311C ＞ T/IVS11-93T ＞ C in this study was found.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Styrenes ; analysis ; Workplace

According to the requirements of the ‘three guidance’ talent-cultivation model pro-posed by our university, humanistic spirit was permeated from multi angels during the process of phys-iology. Students'occupation quality was cultivated; physiology knowledge and clinical practice was combined for students; early contacting with clinical knowledge was conducted and teacher-student role reversal method was applied to train students' communication skills. Teaching practice showed that the reformed teaching model not only can ensure the quality of teaching but also can improve the comprehensive quality of students.

Objective To explore the higher dangerous factors,the early clinical performances and its contents of neonatal pulmonary hemorrhage(NPH).Methods The clinical performances,chest radiograms and autoptical pathological materials of 66 cases of newborns who died of NPH at our neonatal department during 1993 to 2003 were reviewed and analyzed.Results The higher dangerous factors of NPH were premature delivery/low birth weight,serious diseases lead to hypoxia and severe infections.The early clinical performances of NPH were the suddenly aggravation of dyspnea and the increasing of moist sounds.The early X-ray performances were lower penetrance of lung fields extensively and well-distributly with path clouds,the intercostals space usually increased.According to the autoptical(patho)-logy,this X-ray perfomance indicated the edema of the pulmonary with small amount of hemorrhage.Conclusion The patients with the higher dangerous factors and the early clinical performances of NPH,must be diagnosed and interfered it as early as possible to reduce the mortality of NPH.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective To investigate and analyze the postoperative progression of ossification of the patients with ossification of the posterior longitudinal ligament (OPLL) on cervical spine. Methods From Jaunary 2001 to December 2007, 95 postoperative patients with cervical OPLL were followed and analyzed retrospectively. There are 72 males, 23 females, with the average age of 56.3 years (range, 40-73years). The follow-up time was from 1 to 6 years, average 3.1 years. Among them 36 patients were performed with anterior cervical corpectomy, fusion with titanium mesh and fixed with cervical plates, others treated with posterior cervical laminectomy and fixation. 2 of the 95 cases were performed anterior and posterior combined operation. Clinical data, X-rays, CT and MR images and progression of ossification, were measured and analyzed in details .The relationships between the progression of ossification and relative factors, as gender, age, C3 ossified involved, T-OPLL, OPLL-type, time of follow-up, surgical approach, Japanese Orthopaedic Association (JOA) scores and improvement rate of JOA scores, were analyzed. Results Progression of ossification in 39 cases among the 95 followed postoperative OPLL patients, 28 men and 11 women, average age 55.9 years, range 41-71 years. The age of progressed patients included 12 cases of ≤49 years, 12of 50-59 years, 12 of 60-69 years and 3 of ≥70 years. 35 patients were operated by posterior approach and only 4 treated with anterior operation. According to the standard of the progression of ossification that 2 mm in the length or/and thickness, there are 4 cases progressed only in length, 2 only in thickness, other 33 patients both the length and thickness. Progression of length is from 2mm to 20mm (average 7.74±4.71). But thickness is progressed from 2 mm to 6 mm (average 2.67±1.51). From 1 to 3 years follow-up time it appeard as a downtrend about the progression of ossification. But it may appear an uptrend from the 4th year. JOA score and improvement rate of the JOA score were almost improved to the high-point in three years. And according to the statistic data there are obvious relationship between progression of ossification with age, surgical approach and C3 ossified involved. Conclusion There is a high rate of postoperative ossification progression in cervical OPLL patients. Cervical OPLL patients with C3 ossification involved, performed with posterior laminectomy and those young at surgery may have higher rate of progression of the ossification. The JOA score and improvement rate of the JOA score were little influenced by the progression of the OPLL during the short and intermediate-term follow-up.

Objective To compare the clinical therapeutic effects of continuous veno-venous hemofiltration (CVVH) and continuous veno-venous hemodiafiltration (CVVHDF) for treatment of patients with multiple organ dysfunction syndrome (MODS).Methods A prospective observation was conducted, seventy patients with MODS admitted to the Department of Critical Care Medicine of the Affiliated Hospital of Zunyi Medical College from September 2013 to December 2016 were enrolled, and they were divided into a CVVH group and a CVVHDF group according to different treatment, 35 cases in each group. In the CVVH group, the ultrafiltration fluid flow rate was set at 1800 mL/h, while in the CVVHDF group, the flow rate was set at 2000 mL/h for both substitution fluid and dialysate, and the blood flow of the two groups was 150-180 mL/min. The changes of creatinine (SCr), urea nitrogen (BUN), pH value, HCO3-, helper T cell (Th1, Th2) and Th1/Th2 ratio were compared between the two groups before and after treatment for 24, 48 and 72 hours.Results With the extension of time, SCr, BUN were reduced in both groups after treatment compared to those before treatment, 72 hoursafter treatment they reached the lowest value, and the degree of change in CVVHDF group was more significant than that in CVVH group [SCr (μmol/L): 150.62±32.09 vs. 180.41±30.52, BUN (mmol/L): 7.31±2.19 vs. 9.06±2.36. allP < 0.05], after treatment; the pH value, HCO3- level in the two groups had a tendency at first elevated and then lowered, 48 hours after treatment they reached the peak values, compared with those before treatment, the differences were statistically significant [CVVH group: the pH value 7.42±0.08 vs. 7.25±0.10, HCO3- (mmol/L) was 22.49±5.11 vs. 15.65±4.16; CVVHDF group: pH value 7.41±0.04 vs. 7.24±0.11, HCO3- (mmol/L) 23.24±4.78 vs. 15.65±4.16, allP < 0.05], 72 hours after treatment, they began reduced (CVVH group: pH value, HCO3- were 7.39±0.09, 22.35±4.91 respectively, CVVHDF group: pH value, HCO3- were 7.38±0.06, 23.13±4.61 respectively), but they were still significantly higher than those before treatment, and at each time point compared between the two groups, no statistical significant difference was seen (allP > 0.05). With the extension of therapeutic time, Th1, Th1/Th2 in two groups were gradually elevated after treatment, and 72 hours after treatment they reached the peak values; Th2 showed a trend of gradual decrease and after 72 hours of treatment it reached the lowest value, and the changes in CVVHDF group were more significant than those in CVVH group [Th1 (×10-2): 1.51±0.12 vs. 1.21±0.11, Th2 (×10-2): 1.64±0.65 vs. 1.70±0.18, Th1/Th2: 0.91±0.14 vs. 0.71±0.15, allP < 0.05].Conclusion Both CVVH and CVVHDF can effectively remove inflammatory mediators and metabolic products, adjust the balance of acid and base, regulate immune system in the body of patients with MODS, and the CVVHDF is more effective than CVVH.

Objective To investigate the protective effect of sevoflurane on the brain against focal ischemia-reperfusion(I/R)injury and its mechanism.Methods Twenty-four male SD rats weighing 250-300 g were randomly allocated into 3 groups(n=8 each):group Ⅰ sham operation;grouop Ⅱ I/R and group Ⅲ I/R + sevoflurane.The rats were anesthetized with intraperitoneal chloral hydrate 300 mg?kg~(-1).Middle cerebral artery occlusion(MCAO)was produced by insertion of a 4-0 mono-filament nylon thread with rounded tip at bifurcation of right common carotid artery into internal carotid artery.The nylon thread was advanced cranially until resistance was felt.The depth of insertion was 18-20 mm.After 3 h MCAO the thread was withdrawn to allow reperfusion.In group Ⅲ the animals inhaled 1.0 MAC sevoflurane for 30 min at 30 min before reperfusion.The rectal temperature of the animals was kept at 36.5-37.5℃.At the end of 24 h reperfusion the animals were weighed again.The animals'neurological deficit was evaluated using Zea Longa score(0=no defcit,4=unable to walk and unconscious).The animals were then killed.The neuronal apoptosis in striatum was assessed(TUNEL)and the PKC protein expression in striatum was determined by immunocyto-chemistry.Results The body weight of the animals in I/R group was significantly reduced after 24h reperfusion as compared to the body weight before ischemia (P＜0.01),while in control group and sevoflurane group there was no significant difference in the body weight before and after sham operation or I/R.The neurological deficit scores were significantly higher in I/R group than in sevoflurane group.The number of apoptotic neurons in striatum was significantly higher in I/R group than in sevoflurane group.The PKC expression in striatum was significantly higher in sevoflurane group than in I/R group (P＜0.01).Conclusion 1.0 MAC sevoflurane inhalation has protective effect on the brain against I/R injury. Upregulation of PKC expression in striatum decreased by I/R is involved in the mechanism.

Objective To analyze the differential proteomics of ASMC stimulated by wild IL-13 and mutant IL-13 and to investigate the relations of protein profiles of ASMC to asthma and possible targets for the treatment of bronchial asthma.Methods The total proteins of ASMC stimulated by wild IL-13 and mutant IL-13 were separated by immobilized pH gradient(IPG)-based 2-DE and the differentially expressed protein spots were identified by matrix assisted laser desorption-time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE detected approximately(840?21)spots on wild IL-13 samples and(892?17)spots on mutant IL-13 samples(n=3)and(685?19)spots matched.Six significantly differential proteins were subjected to MALDI-TOF-MS analysis and three of them were identified as stathmin 1,Ribosomal protein p~0 and NADH dehydrogenase.Conclusions ASMCs stimulated by wild IL-13 and mutant IL-13 present different proteomic profiles that may shed some light on the mechanism for the asthma causing effect of wild IL-13 and mutant IL-13.