Objective: To investigate the epidemiological and clinical characteristics of 91 cases of dengue fever outbreak in Jiangxi Province in 2019, and to strengthen the management and prevention of dengue fever. Methods: The clinical data, laboratory results and etiology tests of 91 patients with dengue fever from the Ninth Hospital of Nanchang, Zhangshu People′s Hospital, Fengcheng People′s Hospital and Nanchang County People′s Hospital from July 31, 2019 to September 27, 2019 were retrospectively collected. The t test was used for continuous variables and chi-square test was used for categorical variables. Results: A total of 91 dengue fever patients were reviewed. The patients age ranged from 4 to 87 years old, and the majority were adult patients. Most patients were farmers and they all had mosquito-biting history. Local cases were the major source of this outbreak. The incubation time was (5.19±2.96) days. All patients had fever over 38 ℃. Thirty-eight (41.8%) patients developed rashes, mainly distributed on the limbs and trunk, and 15 patients (16.5%) developed myalgia and fatigue. The incidence of rashes in patients admitted in September was 15.9% (7/44), which was significantly lower than 68.1% (32/47) in July and August (χ²=25.262, P<0.01). The incidence of headache in patients admitted in September was 27.3%(12/44), which was significantly lower than 78.7% (37/47) in July and August (χ² =24.206, P<0.01). Among the 91 patients, 64 (70.3%) patients had white blood cell counts less than 4×10⁹/L, 14(15.4%) patients had platelet counts less than 50×10⁹/L. The positive rate of dengue non-structural protein 1 (NS1) antigen was 100.0%(37/37), the positive rates of dengue antibody IgM and IgG were 50.0%(11/22) and 13.6%(3/22), respectively. Among the 113 serum samples tested for dengue virus RNA, 106 were identified with dengue virus type-1 and two were dengue virus type-2. In 22 patients who received tests within 7 days, the positive rate of NS1 antigen was 100.0% (22/22), the IgM and IgG positive rates were 50.0%(11/22) and 13.6% (3/22), respectively, and 19 patients were infected with dengue type-1. Among the IgM antibody positive cases, the onset days before sampling was (5.09±1.64) days, longer than that of IgM antibody negative cases ((2.82±1.83) days), with statistical significance (t=3.063, P=0.007). The onset time before sampling in dengue virus RNA-positive group was (3.53±1.87) days, which was shorter than that of RNA-negative group ((6.67±0.58) days), and the difference was statistically significant (t=2.839, P=0.013). Conclusions The dengue fever outbreak cases in Jiangxi Province in 2019 have typical clinical manifestations of dengue fever. Using the NS1 antigen test, IgM and IgG antibody tests and real-time fluorescent quantitative polymerase chain reaction for dengue virus genotyping during early onset of the disease can assistant clinicians in clinical management, controlling infectious source and stopping disease transmission.

Objective: To evaluate the efficacy and safety of ombitasvir/paritaprevir/ritonavir (OBV/PTV/r) 25/150/100 mg once daily and dasabuvir (DSV) 250 mg twice daily combined with ribavirin in adult patients of Mainland China with chronic HCV genotype 1b infection and compensated cirrhosis. Methods: An open-label, multicenter, phase 3 clinical trial study was conducted in mainland China, Taiwan, and South Korea. Adult patients with compensated cirrhosis (Metavir score =F4) who were newly diagnosed and treated for hepatitis C virus genotype 1b infection with ombitasvir/paritaprevir/ritonavir and dasabuvir combined with ribavirin for 12 weeks were included. Assessed SVR rate of patients obtained at 12 and 24 weeks after drug withdrawal. Efficacy and safety were evaluated in patients who received at least one time study drugs. Results: A total of 63 patients from mainland China were enrolled, 62 of whom (98.4%) had a baseline Child-Pugh score of 5 points. The overall rate of SVR12 and SVR24 in patients was 100% (95% CI: 94.3% to 100.0%). Most of the adverse events that occurred were mild. The incidence of common (≥10%) adverse events and laboratory abnormalities included elevated total bilirubin (36.5%), weakness (19.0%), elevated unconjugated bilirubin (19.0%) and conjugated bilirubin (17.5%), and anemia (14.3%). Three cases (4.8%) of patients experienced Grade ≥ 3 adverse events that were considered by the investigators to be unrelated to the study drug. None patients had adverse events leading to premature drug withdrawal. Conclusion Mainland Chinese patients with chronic HCV genotype 1b infection and compensated cirrhosis who were treated with OBV/PTV/r plus DSV combined with RBV for 12 weeks achieved 100 % SVR at 12 and 24 weeks after drug withdrawal. Tolerability and safety were good, and majority of adverse events were mild.

Objective: To evaluate the safety and efficacy of ombitasvir/paritaprevir/ritonavir (OBV/PTV/r) 25/150/100 mg once daily combined with dasabuvir 250mg, twice daily in non-cirrhotic Chinese adult patients with newly diagnosed and treated chronic HCV genotype 1b infection. Methods: A randomized, double-blind, placebo-controlled, multicenter phase 3 clinical trial was conducted in mainland China, Korea, and Taiwan.Safety and efficacy of OBV/PTV/r plus DSV administered for 12 weeks were evaluated in a newly diagnosed and treated (interferon alpha /pegylated interferon alpha) and ribavirin non-cirrhotic adults with chronic HCVgenotype 1b infection. Patients randomly received OBV/PTV/r plus DSV for 12 weeks (Group A), or placebo for 12 weeks (Group B) followed by an open-label phase of OBV/PTV/r plus DSV for 12 weeks. Sustained response (SVR12) rate obtained at 12 weeks and (SVR24) 24 weeks after discontinuation of treatment, and the incidence of adverse events and laboratory abnormalities after double-blind and open-label phase treatment were assessed. Results: A total of 410 cases of Chinese patients were included and randomly assigned to group A and B (with 205 cases in each group) in a 1:1 ratio. The rates of SVR12 and SVR24 were 99% (95% CI: 94.8% - 99.8%) in the newly diagnosed patients in group A (205 patients) and the rates of SVR12 and SVR24 were 100% in treated patients (95% CI: 96.3% - 100%). Different baseline characteristics had no effect on SVR12 and SVR24 rates. Most of the adverse events occurred were mild, asymptomatic, and≥ 3 laboratory abnormalities during treatment were rare, including elevation of alanine aminotransferase (2 cases in double-blind stage A group), aspartate aminotransferase (Double-blind stage A (3 cases) and total bilirubin (1 case in open-label phase B group); however, those mild adverse events could be recovered after drug withdrawal or discontinuation. only1 person discontinued drugs due to adverse events (Group B, open-label phase). Conclusion The 12 weeks treatment course of OBV/PTV/r combined with DSV produced 99% ~ 100% rates of SVR12 and SVR24 in non-cirrhotic Asian adult patients with newly diagnosed and treated chronic HCV genotype 1b infection, and the tolerance and safety were good.

OBJECTIVE:To establish HPLC fmgerprints of Naodesheng solid dispersion capsules.METHODS:HPLC method was adopted.The determination was performed on Hyspersil ODS2 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelengths were 210 nm (puerarin),345 nm (hydroxfsaffior yellow A).The column temperature was 30 ℃,and sample size was 10 μL.Using puerarin and hydroxysaffior yellow A as reference,HPLC fingerprints of 10 batches of sample were determined.Similarity evaluation,common peak identification and chemical components confirmation were performed by using Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition).RESULTS:There were 29 and 23 common peaks in HPLC fingerprints of 10 batches of samples at 210 nm and 345 nm,respectively.The similarity was higher than 0.90.The medicinal material attribute of common peaks were Panax notoginseng,Ligusticum chuanxiong,Carthamus tinctorius,Pueraia lobata and Crataegus pinnatifida.Moreover,7 chemical components were identified at 210,345 nm,respectively.CONCLUSIONS:Established HPLC fingerprints can provide reference for identification and quality evaluation of Naodesheng solid dispersion capsules.

Objective:To discuss the screening results and clinical characteristics of children of Miao and Dong nationalities with mediterranean anemia in ethnic minority areas of Qiandongnan State of Guizhou Province,and to clarify the differences of the mediterranean anemia among different minorities.Methods:A total of 1 623 children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State were selected by multistage stratified random sampling method;quantitative analysis of HbA2 and HbF was used to screen the selected children with mediterranean anemia initially;phenol chloroform extraction method was applied to extract the DNA from the children with mediterranean anemia;ASO/RDB-PCR reverse dot blot hybridization method was used to analyze the gene characteristics of the children with mediterranean anemia.Results:A total of 1 623 children of Miao and Dong nationalities were selected as the subjects.Among 938 children with Miao nationality,there were 18 children with positive α-mediterranean anemia and 36 children with positive β-mediterranean anemia,and the positive detection rate was 1.92%.Among 685 children with Tong nationality,there were 13 children with positive α-mediterranean anemia and 24 children with positive β-mediterranean anemia,and the positive detection rate was 3.50%.The detection rates of composite of α-and β-mediterranean anemia in the children of Miao nationality and Tong nationality were 1.49% and 4.61%.There was no significant difference in the detection rates of different kinds of mealiterranean anemia between two nationalities (P<0.05).The major gene mutations in α-mediterranean anemia were——SEA/-αα and-α3.7,and the major gene mutations in β-mediterranean anemia were CD17/N and CD14-15/N,while the major gene types of the composite of α-and β-mediterranean anemia were——SEA/β41-42 and——SEA/β17.There was no difference in the positive rates of major gene types of different kinds of mediter ranean anemia between two nationalities(P<0.05).Conclusion:There is no difference in the positive rate of children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State.CD17/N,——SEA/-αα and ——SEA/β41-42 are the major gene types of α-,β-,and αβ-mediterranean anemia,respectively.

Objective To investigate the possible role and significance of soluble programmed death-1 (sPD-1)/soluble programmed death ligand 1 (sPD-L1) in the pathogenesis of oral lichen planus(OLP).Methods Thirty-six patients with OLP(20 cases of reticular OLP and 16 cases of erosive OLP) were enrolled in this study,and 18 healthy people served as controls.Lymphocyte subsets(CD3+,CD4+,CD8+,CD19+,CD16++56+) were examined by flow cytometric analysis and humoral immunity indexes(IgG,IgA,IgM,C3,C4) tested by nephelometry immunoassay.The levels of sPD-1 and sPD-L1 proteins in serum of patients with OLP were determined by enzyme-linked immunosorbent assay.The correlations between the level of sPD-1,sPD-L1 proteins and the immune status and clinical characteristics of patients with OLP were analyzed by SPSS 19.0.Results CD3+,CD4+,CD8+,CD16++56+ in patients with OLP were decreased compared with the normal value,while CD19+ in patients with OLP was increased compared with the normal value(P＜0.05).C3 and C4 in patients with OLP were decreased compared with the normal value,but IgM in patients with OLP was increased(P＜0.05).The levels of sPD-1 and sPD-L1 proteins in patients with OLP were significantly higher than that in control group[26.10(8.81,40.00) ng/L vs 17.65(0.00,26.10) ng/L,29.53 (21.47,36.76) ng/L vs 22.79(1.19,28.29) ng/L](P＜0.05),but the expression of sPD-1 and sPD-L1 was not related with clinical characteristics of OLP.There were negative correlations between the levels of sPD-1 protein and CD4+T cells or CD 16++56+ cells(r1=-0.378,P1=0.007;r2=-0.365,P2=0.009),while there was a positive correlation between the levels of sPD-1 and CD19+B cells(r=0.482,P=0.000).There was a negative correlation between sPD-L1 expression level and CD4 + and a positive correlation between sPD-L1 expression level and IgG(r1=-0.286,P1=0.044;r2=0.365,P2=0.029).Conclusions In patients with OLP,the cellular immune function is low with humoral immunity function disorder.PD-1/PD-L1 signaling pathway,which might be influenced by the involvement of sPD-1 and sPD-L1 proteins in a certain extent,may play an important role in the immune pathogenesis of OLP.

ObjectiveTo investigate the effect of combined spinal epidural analgesia (CSEA) on immune function by observing the changed level of T lymphocyte subsets in maternity sera in labor.MethodsFifty healthy primipara with single birth,vertex present and ASA I between July 2007 and Dec 2007 at the first Affiliated Hospital of Nangchang University,who were in spontaneous labor,were randomly divided into two subgroups when their rerviral dilations were in 2～3 cm.In interfering subgroup( n =25),the puncture point of CSEA was at L3-4 interspace,the fentanyl (20 μg) was used in lumbar anesthesia,the ropivaraine (0.1％) rombined with fentanyl (2 μg/ml) was used in epidural analgesia.Blood samples were taken from the mother vein at cervical dilation in 2 ～ 3 m (T1),fetal disengagement(T2),24 hrs after childbirth ( T3 ).Flow cytometry was used to measure T lymphocyte subsets,Radioimmunoassay (RIA) was used to measure cortisol.In addition,Data on labor progress,VAS score,and neonatal Apgar score were recorded for each patient.Results(1)The active phase in the first stage of labor after analgesia in the CSEA group [ ( 177.64 ± 67.98 ) min ] was significantly lower than that in control group [ (219.40 ± 67.37) min ].No significant difference was found for the active phase in the second stage and the third stage,and for the neonatal Apgar score between the CSEA group [ (32.92 ± 11.59 ) min,( 7.56 ± 2.47 )min,9.20 ± 0.82,respectively ] and the control group [ ( 31.44 ± 13.93 ) min,( 7.28 ± 2.25 ) min,8.84± 1.31,respectively ].(2)The level of cortisol in A group [ ( 548.11 ± 75.67) ng/ml ] was significantly lower than that in C group[ (789.32±96.07) ng/ml] at T2.(3) In two groups,the levels of the CD3+,CD4+,CD4+/CD8 + degraded in different degree at each point,more significantly decreased at T3,and these in C group[ (48.43 ± 6.46) ％,( 31.35 ± 8.93 ) ％,(0.96 ± 0.21 ) ％,respectively ] were significantly lower than those in A group [ (52.3 ± 5.62 ) ％,( 36.90 ± 7.91 ) ％,( 1.16 ± 0.25 ) ％,respectively ].ConclusionsCSEA could shorten the active delivery phase in the first stage of labor,and did not affect the neonatal Apgar score.It can alleviate the inhibitory effect of pain stress response on the immune function.

ObjectiveTo determine the dose of butorphanol at which 50％ and 95％ effective dose (ED50 and ED95) of patients inhibition uterine contraction pain on analgesic artificial abortion.Methods Twenty-six patients undergoing analgesic artificial abortion were sequentially given different doses butorphanol so that the ED50 and ED95 could be determined by up-and-down sequential test.Anesthetic depth was observed when giving propofol with 200 mg/min speed,uterine contraction pain at awaking immediately,10,20 min after awaleing.Data was analyzed by Probit regression analysis for calculating ED50 and ED95..ResultsButorphanol could restrain uterine contraction pain on analgesic artificial abortion and the ED50 was 246 μg with the 95％ confidence interval of 201 μg to 281 μg,the ED95 was 324 μg with the 95％ confidence interval of 287 μg to 548 μg.ConclusionED50 and ED95 value can be expected as a parameter to optimize analgesic artificial abortion.

BACKGROUND: The investigation of culturing, passaging and establishing human limbal stem cells can strengthen the recognition of the stem cells and provide the enough cellular reserve for the basic and clinical research of limbal stem cell transplantation.OBJECTIVE: To explore a method of pessaging and establishing cell line of human limbal stem cells cultured in vitro.DESIGN: Randomized controlled observation.SETTING: Gannan Medical College.MATERIALS: The experiment was performed at Scientific Center of Gannan Medical College and the National Key Laboratory of Ophthalmology Hospital Affiliated to Sun Yat-Sen University from June 2003 to April 2004. Fresh human limbus corneae were isolated from two healthy donors. Procedures were performed according to the informed consent of the donors. Main reagents contained RPMI-1640 (Sigma R8755, containing L-glutamine) and 200 g/L fetal calf serum (FCS) (Gibco 16140-071). DMEM medium, chondroitin sulfatase and human epidermal growth factor (hEGF) were purchased from Sigma Co. USA; HEPES and DMSO were bought from Gibco, USA; 100% glycerinum was purchased from Yunjia Huangpu Pharmaceutical Product Limited Company, PRC; glutaraldehyde was bought from E.Merk, Germany; Alcohol, chlorhydric acid, acetone and methyl aldehyde were purchased from Beijing Chemical Agent Company, PRC; 0.25% parenzyme was bought from Shanghai Xinhua Pharmaceutical Factory, PRC.Above-mentioned reagents were analytical pure grade.METHODS: After digestion, human limbal tissues in limbal basilar part with an abundant pigment were cultured in the culture flask containing RPMI-1640 and 200 g/L FCS and in culture dish containing amniotic extracellular matrix (AECM) as the cultural supporter. Primary and passage cells were observed under light microscope and scanning electron microscope (SEM). The revival ratio of stem cell refrigeration of every generation was calculated by the trypanblau exclusion experiment.MAIN OUTCOME MEASURES: ① Observational results of limbal stem cells during the primary culture and serial subcultivation in vitro, and ② revival ratio of stem cell refrigeration.RESULTS: ①Findings of primary culture: Most limbal stem cells in the culture flask had the adherence and were arrayed uniformly sparsely to form monolayer and adhered to the bottom of culture flask under the inverted phase contrast microscope after 1-day culture. ② Findings of serial subcultivation: After human epidermal growth factor (hEGF) was added into the second passage, cells were scattered into the monolayer and adhered to grow quickly.Morphological variability of all the cells increased obviously when passage the 30th generation. The cellular volume was obviously increasing, and the round or irregular round cells gathered together. The 33rd generation human limbal stem cells still could vigorously differentiate, proliferate and grow in ACEM. ③ The revival ratio of stem cell refrigeration was 82.2%.CONCLUSION: The human limbal stem cell lines were preliminarily established by culturing and freezing the cells of 33 generations in vitro. The human limbal stem cell lines preferred to grow in the culture dish containing AECM as the cultural supporter.

Objective To analyse the effect of mobilization and collection's time of peripheral blood stem cells(PBSC) from 8 cases of healthy donors. Methods The 10 donors were studied by self-control design.The number of aphereses was two times every donors. Healthy donors received rhG-CSF according to two different PBSC collection starting time: group 1:PBSC collection was starts 2 hours(2 h) after the fourth day or the fifth day of rhG-CSF. group 2:PBSC collection was starts 4 hours(4 h) after the fourth day or the fifth day of rhG-CSF.(The first dose of rhG-CSF was given on day 1, considering day 0 as the day before starting mobilization). In this study we have compared with two groups of apheresis product. Results The MNC count was significantly higher for donors 4 h collection (groups 2) then 2 h. ( groups 1)(P