The level of CD4~+ CD25~+ regulatory T cells was assayed in the peripheral blood from the patients with Graves' disease (GD) by floweytometry.CD4~+ CD25~+ regulatory T cells as a kind of non-specific immune cells appear to have no relationship with thyroid function,however,their lowered number in early GD patients suggests that they seem to play an essential role in the pathogenesis of GD.

Objective To establish a content determination method for paeoniflorin in qisheng capsule. Methods The quantitative analysis of paeoniflorin in qisheng Capsule was carried out by high-performance liquid chromatography ( HPLC) . The chromatographic separation was achieved by using a Kromasil C18 chromatographic column (4. 6 mm×250 mm,5 μm) with a mobile phase consisting of methanol,water (1585) at a flow rate of 1. 0 mL·min-1 and 230 nm detection wavelength. Results The linear range was 2. 5-12. 5 μg·mL-1( r =0. 999 9). The average recovery and RSD of the method were 99. 97%and 0. 94%. Conclusion The method is accurate,specific,reproducible,which can effectively be used in quality control of paeoniflorin in qisheng capsule.

This report presented the endemic status of schistosomiasis in the People’s Republic of China at a national level in 2015,and analyzed the data collected from the national schistosomiasis prevention and control system and 457 national schis?tosomiasis surveillance sites. Among the 12 provinces(municipality,autonomous region)endemic for schistosomiasis japonica in P. R. China,5 provinces(municipality,autonomous region),i.e.,Shanghai,Zhejiang,Fujian,Guangdong and Guangxi, had achieved transmission interruption,and 7 provinces of Sichuan,Yunnan,Jiangsu,Hubei,Anhui,Jiangxi and Hunan had achieved transmission control by the end of 2015. There were 453 endemic counties(city,district)covering 252 million peo?ple,specifically including 29 980 endemic villages of 68.61 million people at risk of infection. Among the 453 endemic counties (city,district),75.72%(343/453)and 24.28%(110/453)reached the criteria of transmission interruption and transmission control,respectively. By the end of 2015,it was estimated of 77 194 cases of schistosomiasis,which were decreased by 33.23%compared with 115 614 cases in 2014. No acute schistosomiasis cases were reported in 2015. There were 30 843 advanced schis?tosomiasis cases documented in 2015. A total of 8 736 036 individuals received schistosomiasis examinations and 3 606 individu?als were parasitologically diagnosed,which were decreased by 56.40%compared with 8 270 cases in 2014. An Oncomelania hu?pensis snail survey was performed in 19 965 endemic villages and the snails were found in 5 609 villages,accounting for 28.09%of total villages,with 31 newly detected villages with snails. The snail survey covered an area of 593 572.66 hm2 and snails were found in an area of 173 462.50 hm2,including a newly detected area of 666.04 hm2. No infected snails were found in 2015. A to?tal of 879 373 bovines were raised in the schistosomiasis endemic regions. Of them,526 062 bovines received stool examina?tions,resulting in 315 infected bovines. There were 170 438 schistosomiasis cases receiving drug treatment in 2015,with 2 449 696 individuals undergoing expanded chemotherapy. There were 318 bovines with schistosomiasis receiving drug treatment,with 483 213 bovines undergoing expanded chemotherapy. A total of 144 305.52 hm2 area was subject to snail control by using mollus?cicides,with an actual molluscicide?treated area of 69 221.57 hm2,and 4 572.06 hm2 snail habitats were treated by environmen?tal modification. Based on the data from the 457 national schistosomiasis surveillance sites,the mean Schistosoma japonicum in?fection rate was 0.05% and 0.04% in humans and bovines,respectively. No infected snails were found in all the surveillance sites. The results demonstrate a decline in the endemicity of schistosomiasis in P. R. China and the country reached the criteria of transmission control at the national level. However,the endemic situation of schistosomiasis is unstable in some regions with newly?reached transmission control,and further control and effective surveillance should be strengthened to consolidate the achievements and reduce the endemic situation of schistosomiasis in P. R. China.

Objective To discuss the diagnostic value of high resolution computed tomography (HRCT)and MRI in the congenital inner ear malformation incomplete partition type Ⅱ deformity(Mondini deformity)combined with spontaneous cerebrospinal fluid (CSF)otorrhea.Methods The imaging data including the multiplanar reformation(MPR)images of HRCT,curved planar reforma-tion(CPR)images and MR hydrography images in three patients complicated with recurrent meningitis were retrospectively ana-lyzed.The anatomic changes of the inner ear and middle ear on the affected side were observed,the clinical and imaging features of this disease were summarized.Results One patient had bilateral incidence of the incompletely separated inner ear,combined with semicircular canal dysplasia and cochlear pipe dilation.Two patients had unilateral onset,one of them had concurrent facial nerve ca-nal dysplasia.Humble stapes floor and the defect of the bottom of the internal auditory canal were seen in all the three patients.Con-clusion HRCT with post-processing technology,and MRI have great significance in the diagnosis of the congenital inner ear malfor-mation incomplete partition type Ⅱ deformity (Mondini deformity)combined with spontaneous CSF otorrhea.

OBJECTIVE:To explore the methods of clinical pharmacists providing pharmaceutical care for a patient with N-methyl-D-aspartate receptor(NMDAR)encephalitis complicating with multiple organ infections. METHODS:Taking one clinical case as breakthrough point,the points of pharmaceutical care provided by clinical pharmacists for NMDAR encephalitis complicat-ing with multiple organ infections were analyzed,so as to put forward the suggestions in the field of antibiotics selection,medica-tion approach based on pharmacokinetics,ADR disposal and nutrition support. RESULTS:Clinical pharmacists applied pharmaceuti-cal care to resolve ADR as abnormal liver enzyme timely,and the symptom had been improved gradually. Then the patient was dis-charged from the hospital. CONCLUSIONS:Clinical pharmacists provide pharmaceutical care and screen the possible risk of drug use to avoid the occurrence of severe ADR.

Objective To analyze the investigation data of the national schistosomiasis surveillance sites in 2015,so as to provide scientific evidences for schistosomiasis control,elimination and surveillance. Methods According to National Schisto-somiasis Surveillance Programme(version 2014),457 surveillance sites were selected,and the investigation data in residents, floating population,domestic animals and Oncomelania hupensis snails were collected and analyzed from four types of endemic counties. Results A total of 4468 seropositive cases were detected from 133350 residents,among which 4457 residents with seropositive results received the etiological tests,and 71 of them were identified with positive results. Most of them were fisher-men and farmers in the middle and old-aged group. The schistosomiasis infection rate was 0.05% in local population. Totally 977 seropositive cases were examined from 85047 migrant individuals,and 16 positive cases were found out from 966 individuals who took etiological tests,which showed the schistosomiasis infection rate was 0.02% in floating population. Imported cases were found among floating people in four provinces,namely Zhejiang,Hunan,Hubei and Anhui provinces. No acute schistoso-miasis cases were reported. A total of 13406 head of cattle received examinations and only 5 were determined as stool positives. The cattle infection rate was 0.04%. The snail survey covered an area of 22295.13 hm2 and snails were found in an area of 7426.63 hm2,including 3.47 hm2 newly detected area with snails. No schistosome-infected snails were found. Conclusions Based on the collected data from 457 national schistosomiasis surveillance sites of China,the Schistosoma japonicum infection rate is 0.05% in local population which maintains a stably descending trend. In floating population,there are imported schistosome-in-fected persons. Cattle are still a vulnerable species infected with schistosome. Although no infected snails are found,snails are widely distributed in endemic areas. Some provinces detect areas with snails for the first time or the reproduction of snails. The staff in endemic provinces should carry out the surveillance work according to National Schistosomiasis Surveillance Programme (version2014)to improve the surveillance system,and enhance the sensibility and effectiveness of surveillance work.

Objective:To investigate the relationship of urinary 8-oxoguanosine(8-oxoGsn)with muscle mass, muscle strength, advanced glycation end products(AGEs), trace elements, heavy metals and other health-related indictors in different age groups of the Beijing area.Methods:A cross-sectional research was conducted.Healthy adults aged 25 to 93 years who sought health examination in the Health Examination Center of Beijing Hospital were recruited.Participants were divided into the young and middle-aged group and the elderly group with age 60 as the cutoff.Urinary 8-oxoGsn levels were detected by mass spectrometry and adjusted using urine creatinine values.Body composition was measured by multifrequency bioelectrical impedance analysis(BIA). Grip strength, 6-meter walking speed and 5-times sit to stand test were conducted by experienced team members.Skin autofluorescence was used to detect skin AGEs.A portable optical scanner was used to detect heavy metals and trace elements using reference points of the palm.Levels of fasting blood glucose, glycosylated hemoglobin, high-density lipoprotein and other common blood biochemical indicators were measured.Results:A total of 106 subjects were enrolled, including 68 in the young and middle-aged group and 38 in the elderly group.The proportion of patients with hypertension(14 ases or 36.8% vs.7 ases or 10.3%), systolic blood pressure[130(120, 140) vs.120(110, 126)mmHg], fasting blood glucose[5.7(5.2, 5.9)mmol/L vs.5.2(4.9, 5.5)mmol/L], glycosylated hemoglobin[6.0(5.7, 6.2)% vs.5.7(5.4, 5.9)%], 8-oxoGsn/Cre[1.9(1.4, 2.6) vs.1.3(1.0, 1.6)], AGEs(2.44±0.46 vs.2.01±0.29), 5-times sit to stand test scores[7.8(6.9, 9.8)s vs.6.0(5.0, 6.8)s], magnesium(31.4±7.2 vs.27.7±6.4), mercury(0.013±0.003 vs.0.008±0.003)and silver[0.011(0.010, 0.012) vs.0.010(0.009, 0.011)]were higher in the elderly group than in the young and middle-aged group, while grip strength[28.0(22.0, 35.1)kg vs.36.6(28.5, 49.1)kg], fat-free mass[44.9(37.5, 51.1)kg vs.53.3(42.4, 58.5)kg], trunk muscle mass[21.0(17.5, 23.9)kg vs.25(19.8, 27.4)kg], appendicular skeletal muscle mass[20.9(17.6, 23.9)kg vs.24.9(19.8, 27.3)kg], calcium[273.3(219.1, 480.0) vs.457.8(428.5, 489.1)], cobalt[0.029(0.027, 0.031) vs.0.031(0.028, 0.034)], selenium[1.44(0.93, 1.71) vs.1.61(1.53, 1.68)]and nickel[3.5(3.3, 4.0)*10 -3vs.3.8(3.6, 4.1)*10 -3]were lower in the elderly group than in the young and middle-aged group( P<0.05). Urinary 8-oxoGsn/Cre levels were positively correlated with age, time of 5-times sit to stand test, AGEs, fasting blood glucose, mercury and aluminum( rs=0.443, 0.292, 0.357, 0.205, 0.316 and 0.214, P<0.05), and negatively correlated with trunk muscle mass, appendicular skeletal muscle mass, fat-free mass, grip strength, silicon and manganese( rs=-0.334, -0.333, -0.332, -0.366, -0.246 and -0.234, P<0.05), with statistical significance. Conclusions:Increased urinary 8-oxoGsn/Cre levels are correlated with decreased muscle mass, poor physical function, accumulation of AGEs, decreased trace element levels and increased heavy metal levels.Therefore, 8-oxoGsn has the potential to be a broadly representative and sensitive indicator for health assessment.

Objective:To establish an LC-MS/MS method for the content determination of triptolide in Shenshe ointment.Methods:The chromatographic separation was performed on an Agilent TC-C18 (250 mm×4.6 mm,5 μm)column with the mobile phase consisting of methanol-0.1% formic acid solution (73∶27,v/v).The flow rate was 0.5 ml·min-1 and the column temperature was maintained at 35℃.An electrospray ionization (ESI) source was applied with multiple reaction monitoring (MRM) combined with a positive mode.Metronidazole was used as the internal standard.The mass transitions were 361.4→43.2 for triptolide and 172.1→128.0 for metronidazole,respectively.Results:The linear range was 10-500 ng·ml-1 with good correlation coefficient (r=0.999 9).The limit of quantification (LOQ) was 9.5 ng·ml-1.The intra-and inter-day RSDs of peak areas were all less than 3% and the average recovery was 96.87%(RSD=2.79,n=6).Conclusion:The established LC-MS/MS method is simple,efficient,sensitive and accurate in the quality control of Shenshe ointment.

OBJECTIVETo gain insights into the role of forkhead box protein 3 (Foxp3) in the pathogenesis of hepatocellular carcinoma (HCC) by performing a comparative analysis of Foxp3 mRNA expression and promoter methylation status in HCC and normal liver tissues.

METHODSThirty-nine HCC and 13 normal liver tissue specimens were evaluated by real-time quantitative PCR and pyrosequencing to measure the expression of Foxp3 mRNA and determine the methylation status of its promoter, respectively. Statistical analyses of the data were conducted by rank-sum test and Spearman's rank correlation coefficient test.

RESULTSThe HCC specimens showed significantly higher mRNA expression of Foxp3 (vs. normal liver tissues, Z =-2.770, P =0.0056). Moreover, the HCC specimens showed significant hypomethylation of the Foxp3 promoter site A (vs. normal liver tissues, Z =2.118, P =0.0339), and the Foxp3 mRNA level was negatively correlated with the methylation of site A (rs =-0.344, P =0.046). None of the other four sites in the Foxp3 promoter showed a significant difference in methylation, and the overall methylation was not significantly different between the HCC and normal liver tissues.

CONCLUSIONOverexpression and low methylation of Foxp3 may be involved in the oncogenic and progression processes of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; CpG Islands ; DNA Methylation ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Promoter Regions, Genetic ; RNA, Messenger ; genetics

Recombineering, a new genetic engineering technology based on high efficiency in vivo homologous recombination, can be used in target DNA knock-in, knock-out and gene cloning. In the process of gene subcloning mediated by Recombineering technique, high-quality target DNA fragments were difficult to obtain using in vitro overlapping PCR,therefore the efficiency of in vivo homologous recombination was severely interrupted. To solve this problem, some technology improvements have been established based on the principle of Red recombinases. The PCR DNA fragments of egfp and kan genes with complementary sequences on the end of each fragment were co-introduced into a pcDNA3.1 vector and Red recombinases containing E. coli DY331 host cells by electroporation. A recombinant plasmid pcDNA3.1-egfp-kan was screened directly by antibiotic marker. The positive rates can reach to 45%. The EGFP gene expression of pcDNA3.1-egfp-kan can be observed by transient transfection of 293 eukaryotic cells.
Bacteriophage lambda ; enzymology ; genetics ; DNA, Recombinant ; genetics ; Electroporation ; Escherichia coli ; genetics ; Gene Fusion ; genetics ; Genetic Engineering ; methods ; Green Fluorescent Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Recombinases ; genetics ; metabolism ; Recombination, Genetic