OBJECTIVE: To establish an HPLC method for simultaneous determination of the content of sulfadiazine and dexamethasone acetate in compound nasal ointment.METHODS: The determination was performed on Inertsil C8-3 column with column temperature set at 40 ℃;the mobile phase consisted of methanol-acetonitrile-water(72∶3∶25) with a flow rate of 1.0 mL?min-1.The detective wavelength was set at 240 nm and the sample size was 20 ?L.RESULTS: The linear ranges for sulfadiazine and dexamethasone acetate were 74.65～746.5(r=0.999 9) and 1.495～14.95 ?g?mL-1(r=0.999 7),respectively.The average recoveries were 100.72%(RSD=1.7%) and 100.41%(RSD=1.6%),respectively.CONCLUSION: This method is rapid,simple,accurate and reliable and suitable for the determination of the content of sulfadiazine and dexamethasone acetate in compound nasal ointment.

Objective:To investigate the clinical characteristics of patients infected with Brucella Melitensis ( B. Melitensis) type 1 and type 3 in Hulunbuir, Inner Mongolia Autonomous Region. Methods:A retrospective analysis method was used to collect clinical medical records of patients with brucellosis admitted to Hulunbuir People's Hospital from June 2013 to August 2017, and 71 patients with brucellosis positive in blood culture and identified by polymerase chain reaction (PCR) and AMOS-PCR were selected as the study subjects. According to the identification results, they were divided into B. Melitensis type 1 and type 3 groups. General information, epidemiological characteristics, clinical characteristics, laboratory examinations, complications and efficacy of the two groups were compared. Results:Among 71 patients with brucellosis, 22 cases were B. Melitensis type 1, including 16 males and 6 females, aged (39.91 ± 16.04) years old; 49 cases were B. Melitensis type 3, including 34 males and 15 females, aged (40.67 ± 18.72) years old. There were no significant differences in gender composition and age between the two groups (χ 2 = 0.081, t = 0.166, P > 0.05). There were 10 cases(45.5%) of B. Melitensis type 1 patients living in agricultural areas, 10 cases (45.5%) in pastoral areas, and 2 cases (9.1%) in cities; there were 40 cases (81.6%) of B. Melitensis type 3 patients living in agricultural areas, 7 cases (14.3%) in pastoral areas and 2 cases (4.1%) in cities, and the difference between regions was statistically significant (χ 2 = 9.276, P < 0.05). Testicular swelling and pain symptoms [22.7% (5/22), 6.1% (3/49)] in B. Melitensis type 1 and type 3 patients were compared, the difference was statistically significant (χ 2 = 4.187, P < 0.05); other clinical features were compared, the differences were not statistically significant ( P > 0.05). There were no significant differences in white blood cell count (WBC) and platelet count (PLT) decreased, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) increased between the two groups ( P > 0.05). The number of complications in B. Melitensis type 1 and type 3 patients were 12 cases (54.5%) and 14 cases (28.6%), respectively, and the difference between the two groups was statistically significant (χ 2 = 4.413, P < 0.05). Ten cases (45.5%) of B. Melitensis type 1 patients were cured, 12 cases (54.5%) were improved, 34 cases (69.4%) of B. Melitensis type 3 patients were cured, 15 cases (30.6%) were improved, and there were no invalid or relapsed patients in both groups, the difference in curative effect between the two groups was statistically significant (χ 2 = 3.690, P < 0.05). Conclusions:In Hulunbuir, Inner Mongolia Autonomous Region, there are differences in the living areas of B. Melitensis type 1 and type 3 patients. The B. Melitensis type 1 patients are prone to testicular swelling and pain and brucellosis complications.

Objective To investigate the associations of cognitive function with apolipoprotein E (APOE) gene polymorphism and chronic diseases among long-lived people in Zhongxiang City of Hubei Province.Methods A total of 110 long-lived elderly residents aged 90 years and over were collected.Their cognitive function was determined face-to-face using questionnaires by trained interviewers.According to mini-mental state examination(MMSE) scores,subjects were divided into a no dementia risk group and a high dementia risk group.General demographic characteristics,activities of daily living,depression state and nutrition status were compared between the two groups.Correlations of dementia with APOE gene polymorphism and chronic diseases were analyzed.Results The average MMSE score was 22.3±4.8.Among the 110 long-lived people,18 cases had a high risk for dementia,accounting for 16.4％,and 92 cases had no risk of dementia,accounting for 83.6％.The risk of dementia in long-lived elderly people was correlated with activities of daily living,mental state,nutritional status and falls(all P＜0.05).There were 8 cases with the APOE gene ε4/ε3 genotype in the high dementia risk group and 16 cases with the APOE gene ε4/e3 genotype in the no dementia risk group,with the former group showing a higher frequency of the APOE ε4/ε3 genotype (44.4％ vs.17.4％,x2 =6.46,P＜0.05).The former group also seemed to have a higher APOE ε4 frequency,but the difference was not statistically significant(22.2 ％ vs.10.3 ％,x2 =3.96,P =0.055)Chronic diseases prevalent in the long-lived people were hypertension(86 cases,78.2 ％),hearing loss (72 cases,65.5％),hyperlipidemia(56 cases,50.9％),anemia(43 cases,39.1％),impaired vision(39 cases,35.5 ％),chronic kidney diseases(25 cases,22.7 ％),chronic heart diseases (18 cases,16.4 ％) and osteoarthritis(18 cases,16.4％).No correlation was found between the risk of dementia and chronic diseases(P＞0.05).Conclusions Cognitive function is highly correlated with activities of daily living,mental state and nutritional status among long-lived elderly people in Zhongxiang City.The risk of dementia has a correlation with the APOE gene e4/e3 genotype but not with chronic diseases in long-lived people.

Objective To evaluate the influence of Danhong injection on endothelial progenitor cells ,inflammatory factor and neural function in patients with acute cerebral infarction .Methods 116 cases of patients with acute cerebral infarction in our hospi-tal from April 2016 to November 2016 were chosen for the research ,and were divided into research group (Danhong injection with routine treatment) and control group (routine treatment) ,the changes of endothelial progenitor cells ,neural function and inflamma-tory factors between two group were compared 30 d after treatment .Results Endothelial progenitor cells in peripheral blood be-tween two group had no statistically significant difference before treatment (P>0 .05) .After treatment ,endothelial progenitor cells of two group all increased ,and had a statistically significant difference compared with before (P< 0 .05) .Endothelial progenitor cells at 7 d ,14 d and 30 d after treatment of research group were higher than those of control group ,and the difference was statisti-cally significant (P< 0 .05) .NIHSS score before treatment between two group had no statistically significant difference (P>0 .05) .After treatment ,NIHSS score of two group all decreased ,and had statistically significant difference compared with before (P<0 .05) .NIHSS score at 7 d ,14 d and 30 d after treatment of research group were higher than those of control group ,and the difference was statistically significant (P<0 .05) .IL-6 and IL-10 levels before treatment between two group had no statistically sig-nificant difference (P>0 .05) .After treatment ,IL-6 and IL-10 levels of two group all decreased ,and had a statistically significant difference compared with before (P<0 .05) .IL-6 and IL-10 levels at 7 d and 14 d after treatment of research group were higher than those of control group ,and the difference was statistically significant (P<0 .05) .Conclusion Danhong injection could effec-tively improve the level of endothelial progenitor cells and inflammatory ,and promote the recovery of neural function .

Objective:To explore the relationship between the Spike and the Gamma rhythm of the local field potential (LFP) in Alzheimer's disease (AD) transgenic mice during fear memory activity.Methods:Six-month-old APP/PS1/tau three transgenic (3xTg) mice and wild-type (WT) mice were divided into 3xTg group and WT group, with 10 mice in each group. The electrodes were embedded into the hippocampus of mice under sterile conditions, and the behavioral experiment of conditioned fear box test was carried out two weeks later. The changes of Gamma rhythm, Spike and Burst firing were recorded and analyzed by the wireless telemetry device which embedded in the mouse head. Finally, the correlation between Gamma rhythm and Spike was calculated by entropy value.Results:(1) In behavioral experiments, the freezing ratio caused by conditioned stimulation (CS) in 3xTg mice was ((54.07±2.32)%), which was significantly lower than that of WT mice ((76.21±2.88)%) ( t=4.796, P<0.01). (2) Simultaneously recorded the average power of the Gamma oscillation in the Pre-CS period of the WT mice was ((11.574±1.147) dB), which increased to ((18.108±1.177) dB) after CS ( t=3.386, P<0.01). After CS administration, the average power of Gamma in 3xTg group((12.346±1.345) dB) was significantly lower than that of WT group ( t=3.423, P<0.01). (3) The frequency of Spike release in WT mice during the Pre-CS period was ((5.667±1.475)times/s), significantly increased to ((11.008±1.335) times/s) after CS ( t=3.542, P<0.01). The frequency of Spike release of 3xTg mice after CS ((5.249±1.033) times/s) was significantly lower than that of WT group ( t=4.788, P<0.01). (4) The Burst duration of WT group in pre-CS and CS period were ((0.550±0.043)s) and ((1.075±0.034)s), respectively. It suggested that the Burst firing frequency of WT group increased significantly after conditional stimulation ( t=5.188, P<0.01). However, the release interval of 3xTg group after CS ((0.619±0.033)s) was significantly lower than that of WT group ( t=3.352, P<0.01). (5) After CS, the Spike-Gamma entropy curve of WT mice was always higher than that of 3xTg mice. The maximum correlation of WT group and 3xTG group were (0.403±0.031) and (0.314±0.028), respectively. The Spike-Gamma correlation of the 3xTg group was significantly lower than WT mice ( t=3.372, P<0.01). Conclusion:The defect of fear memory in Alzheimer's disease may be caused by the disharmony of Spike-LFP (Gamma) distribution.

OBJECTIVE: To carry out phenotypic and genotypic analysis for two Chinese pedigrees affected with coagulation factor XII (F XII) deficiency. METHODS: Plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), and blood coagulation factor VIII, IX, XI, XII activity (FVIII:C, FIX:C, FXI:C, FXII:C) were determined with one stage clotting assay on a STAGO coagulation analyzer. FXII antigen was determined with an enzyme linked immunosorbent assay (ELISA). The 14 exons and their flanking sequences of the F12 gene were subjected to PCR amplification and Sanger sequencing. The conservation and structure of mutant protein were analyzed with MegAlign software and PYMOL software. RESULTS: The APTT of the probands was significantly prolonged, while their FXII:C and FXII:Ag were significantly reduced. Genetic analysis of the proband has revealed three novel mutations in the F12 gene, including g.5972G>A splice site mutation in intron 5, g.8810_8814delGTCTA in exon 14, and g.6259G>A (p.Pro182Leu) in exon 7. In addition, a previously known mutation IVS13-1G>A has been found. CONCLUSION Four mutations have been identified in the two Chinese pedigrees, among which three were novel. Above mutations probably played a role in the defect of FXII in the two pedigrees.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Genetic Testing ; Humans ; Pedigree

OBJECTIVETo explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with FX deficiency.

METHODSProthrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen, FII activity(FII:C), FVII activity(FVII:C), FIX activity (FIX:C), FX activity(FX:C) were determined with a one-stage clotting assay. The FX antigen(FX:Ag) was detected with an enzyme linked immunosorbent assay(ELISA). The 8 exons, introns and 5' and 3' untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing. Suspected mutation was confirmed by reverse sequencing. Polymorphisms were excluded by direct sequencing of 100 healthy individuals.

RESULTSThe PT and APTT of the proband have prolonged to 16.1 s and 49.0 s, respectively. Her FX:C and FX:Ag were reduced by 27% and 56%, and her mother's PT, APTT, FX:C and FX:Ag were 14.8 s, 37.4 s, 44%, 34%, respectively. Her grandmother's PT, APTT, FX:C and FX:Ag were 15.8 s, 42.2 s, 31%, 45%, respectively. The results of her father and other family members were all within the normal range. Genetic analysis has revealed a heterozygous G to A mutation in the proband at position 28076 in exon 8 of the F10 gene, which resulted in a p.Gly363Ser substitution. The same mutation was also found in her mother and grandmother. No mutation of the F10 gene was found in her father. Gly363Ser may result in changes in the secondary structure of the FX protein and reduction of its activity.

CONCLUSIONThe g.28076G to A(p.Gly363Ser) mutation of the F10 gene probably underlies the FX deficiency in this pedigree. The mutation was discovered for the first time in Chinese patients.

Objective:To analyze the epidemiological characteristics, clinical features of brucellosis patients with abnormal blood routine, and to improve the awareness of brucellosis among clinicians.Methods:A total of 1 036 patients with brucellosis admitted to the Department of Brucellosis, Hulunbuir People's Hospital from January 2011 to December 2017 were selected, including 274 patients with abnormal blood routine (case group), and 762 patients with normal blood routine(conrtol group). Epidemiological characteristics, clinical features and laboratory tests were analyzed retrospectively.Results:In 274 patients of case group, there were 128 males and 146 females, and the age was (36.3 ± 18.7) years old. In 762 patients of conrtol group, there were 381 males and 381 females, and the age was (35.4 ± 20.4) years old, and there were no significant differences in sex ratio and age between the two groups ( P > 0.05). The main route of infection in the two groups was to raise livestock such as sheep and cattle, 254 cases (92.7%) and 724 cases (95.0%), respectively. The proportions of patients with dizziness symptoms were 31.0% (85/274) and 17.7% (135/762) in the two groups, and the proportions of patients with rash were 14.2% (39/274) and 3.0% (23/762), and the differences were statistically significant between the two groups (χ 2 = 21.331, 45.054, P < 0.05). The symptoms of fever, sweating and fatigue were common in both groups, and the splenomegaly was the most common sign. However, there were no significant differences in the proportion of abnormal characteristics between the two groups ( P > 0.05). Among the 274 patients in case group, 48 had leucopenia, 160 had anemia, and 148 had thrombocytopenia; and 17 had both leucopenia and anemia, 23 had both leucopenia and thrombocytopenia, and 16 had both anemia and thrombocytopenia, and 13 had leucopenia, anemia, and thrombocytopenia at the same time. Conclusions:In the brucellosis epidemic area, when the patient has symptoms such as fever, fatigue, sweating, leucopenia, anemia, and thrombocytopenia, and excluding abnormal blood routine caused by other reasons, clinicians should consider the possibility of Brucella infection.

BACKGROUND:Oxidative injury is considered to be one of the important factors of cerebral ischemia-reperfusion injury.Superoxide dismutase 2(SOD2)is a key mitochondrial antioxidant molecule,and fenofibrate can regulate the expression of SOD2 by activating peroxisome proliferator-activated receptor α. OBJECTIVE:To explore whether the mechanism of fenofibrate in the treatment of cerebral ischemia-reperfusion injury depends on the expression of SOD2. METHODS:The TALENs system was used to construct SOD2 transgenic mice.The transgenic mice were genotyped by PCR and DNA sequencing techniques.The expression of SOD2 protein in transgenic mice was detected by western blot assay.Wild-type and SOD2 transgenic mice were randomly divided into four groups:wild-type control group(n=6),wild-type fenofibrate group(n=6),SOD2 transgenic control group(n=5)and SOD2 transgenic fenofibrate group(n=5).A mouse model of middle cerebral artery occlusion was prepared using the suture-occlusion method.After 90 minutes of ischemia,the thread was removed to reperfuse cerebral blood flow for 30 minutes.A cerebral blood flow monitor was used to monitor local cerebral blood flow.Brain tissue slices were taken for 2,3,5-triphenyltetrazolium chloride staining to analyze the situation of cerebral infarction in each group. RESULTS AND CONCLUSION:After PCR and DNA sequencing analysis,nine SOD2+/+ transgenic mice were successfully constructed.After cerebral ischemia-reperfusion,the wild-type fenofibrate group showed partial recovery of cerebral blood flow and significantly reduced cerebral infarction volume compared with the wild-type control group(P<0.001).There was no significant difference in cerebral blood flow and cerebral infarction volume between the SOD2 transgenic fenofibrate group and the SOD2 transgenic control group.The SOD2 transgenic control was superior to the wild-type control group in terms of improving cerebral blood flow and cerebral infarction(P<0.001).There were also no significant differences in cerebral blood flow and cerebral infarction volume between the wild-type fenofibrate group and the SOD2 transgenic control group and between the wild-type fenofibrate group and the SOD2 transgenic fenofibrate group.To conclude,the expression of SOD2 is one of the mechanisms of fenofibrate in the treatment of cerebral ischemia-reperfusion injury.

Objective:To evaluate the effect of controlled low central venous pressure with milrinone on laparoscopic hepatectomy in the patients.Methods:Fifty American Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both sexes, aged 18-64 yr, with body mass index of 18-30 kg/m 2, of Child-Pugh grade A or B, undergoing elective laparoscopic hepatectomy, were divided into 2 groups ( n=25 each) using a random number table method: milrinone group (group M) and nitroglycerin group (group NG). After the start of surgery, milrinone 0.5 μg·kg -1·min -1 was continuously infused in group M, and nitroglycerin was continuously infused with the initial dose of 0.5 μg·kg -1·min -1 to maintain central venous pressure (CVP)≤5 mmHg in group NG.Mean arterial pressure and heart rate were recorded on admission to the operation room (T 0), at skin incision (T 1), at the beginning of liver resection (T 2), at completion of liver resection (T 3), at the end of operation (T 4), and CVP, cardiac index and stroke volume variation were recorded at T 1-4.Internal jugular vein blood samples were collected to determine the concentrations of hemogloblin, blood lactate at T 1 and T 4, and serum alanine aminotransferase, aspartate aminotransferase and creatinine concentrations at 1, 3 and 7 days after surgery.The score of blood oozing in hepatic surgical field, amount of norepinephrine used, blood loss, postoperative recovery and occurrence of complications within 7 days after operation were recorded. Results:Compared with group NG, cardiac index was significantly increased at T 2, 3, the CVP was decreased at T 2, the blood oozing score, blood loss, consumption of norepinephrine, and concentrations of blood lactate were decreased, and the postoperative drainage indwelling time was shortened in group M ( P<0.05). There was no significant difference in the serum alanine aminotransferase, aspartate aminotransferase and creatinine concentrations and incidence of postoperative complications at 1, 3 and 7 days after operation between the two groups ( P>0.05). Conclusions:Milrinone is better than nitroglycerin in decreasing central venous pressure, reducing blood loss, maintaining stable circulatory function and tissue perfusion in laparoscopic hepatectomy.