DNA methylation of skeletal muscle function-related secretary factors identifies FGF2 as a potential biomarker for sarcopenia.

BACKGROUND: Sarcopenia is characterized by progressive loss of muscle mass and function due to aging. DNA methylation has been identified to play important roles in the dysfunction of skeletal muscle. The aim of our present study was to explore the whole blood sample-based methylation changes of skeletal muscle function-related factors in patients with sarcopenia. METHODS: The overall DNA methylation levels were analysed by using MethlTarget™ DNA Methylation Analysis platform in a discovery set consistent of 50 sarcopenic older adults (aged ≥65 years) and 50 age- and sex-matched non-sarcopenic individuals. The candidate differentially methylated regions (DMRs) were further validated by Methylation-specific PCR (MSP) in another two independent larger sets and confirmed by pyrosequencing. Receiver operating characteristic (ROC) curve analysis was used to determine the optimum cut-off levels of fibroblast growth factor 2 (FGF2)_30 methylation best predicting sarcopenia and area under the ROC curve (AUC) was measured. The correlation between candidate DMRs and the risk of sarcopenia was investigated by univariate analysis and multivariate logistic regression analysis. RESULTS: Among 1149 cytosine-phosphate-guanine (CpG) sites of 27 skeletal muscle function-related secretary factors, 17 differentially methylated CpG sites and 7 differentially methylated regions (DMRs) were detected between patients with sarcopenia and control subjects in the discovery set. Further methylation-specific PCR identified that methylation of fibroblast growth factor 2 (FGF2)_30 was lower in patients with sarcopenia and the level was decreased as the severity of sarcopenia increased, which was confirmed by pyrosequencing. Correlation analysis demonstrated that the methylation level of FGF2_30 was positively correlated to ASMI (r = 0.372, P < 0.001), grip strength (r = 0.334, P < 0.001), and gait speed (r = 0.411, P < 0.001). ROC curve analysis indicated that the optimal cut-off value of FGF2_30 methylation level that predicted sarcopenia was 0.15 with a sensitivity of 84.6% and a specificity of 70.1% (AUC = 0.807, 95% CI = 0.756-0.858, P < 0.001). Multivariate logistic regression analyses showed that lower FGF2_30 methylation level (<0.15) was significantly associated with increased risk of sarcopenia even after adjustment for potential confounders including age, sex, and BMI (adjusted OR = 9.223, 95% CI: 6.614-12.861, P < 0.001). CONCLUSIONS: Our results suggest that lower FGF2_30 methylation is correlated with the risk and severity of sarcopenia in the older adults, indicating that FGF2 methylation serve as a surrogate biomarker for the screening and evaluation of sarcopenia.

Association of AST/ALT (De Ritis) ratio with sarcopenia in a Chinese population of community-dwelling elderly.

Background: The aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio, also known as De Ritis ratio, has been reportedly associated with malnutrition which plays a crucial role in sarcopenia. The aim of this study was to examine the relationship between AST/ALT ratio and sarcopenia in the Chinese community-dwelling elderly. Methods: A cross-sectional study with 2751 participants (1343 men and 1408 women) aged ≥60 years was performed. Appendicular skeletal muscle mass index (ASMI), grip strength, and gait speed were measured to diagnose sarcopenia according to the latest Asian Working Group for Sarcopenia (AWGS) consensus. The association of AST/ALT ratio with sarcopenia was examined using logistic regression analysis. Results: The prevalence of sarcopenia in the present study was 4.4%. AST/ALT ratio was higher in the sarcopenia group than in the non-sarcopenia group (1.30 ± 0.33 vs. 1.16 ± 0.62, P = 0.010). AST/ALT ratio was negatively correlated with the components of sarcopenia, including ASMI, grip strength, and gait speed. Logistic regression analysis indicated that high AST/ALT ratio (>1.20) was associated with increased risk of sarcopenia even after adjustment for potential confounders (adjusted OR = 2.33, 95%CI = 1.48-3.68, P < 0.001). Stratification analyses indicated that the association of high AST/ALT ratio with high risk of sarcopenia was more significant in males and the elderly with ≥70 years. Conclusions: Our findings demonstrate that high AST/ALT ratio is associated with increased risk of sarcopenia in a Chinese population of community-dwelling elderly.

[Epidemiological Survey on Intestinal Parasitic Infections in Lhasa City in 2014].

Objective: To investigate the prevalence of parasitic infections in Lhasa, Tibet. Methods: The residents in Chengguan District, Dangxiong County, Dazi County and Qushui County were selected to participate in the study in 2014, using the stratified random sampling method. Infections with the soil-borne nematodes in feces were examined in triplicates using the Kato-Katz method. Further, the Enterobius vermicularis were examined using the cellophane anal swab method in children aged 3-12 years, and the intestinal protozoa were detected by iodine staining. The parasitic infection was analyzed by sex, age, and occupation. Comparisons were made using Chi-square test. Results: A total of 1 015 residents（39.3±18.2 years: age range, 1-89 years） participated in the study, including 391 males（38.6%） and 623 females（61.4%）. Two hundred and forty-nine participants were positive for parasitic infections, with an overall infection rate of 24.5%（249/1 015）. The infection rate was 21.7%（220/1 015） for Taenia sp., 0.2%（2/1 015） for Trichuris trichiura and E. vermicularis respectively, 4.1%（42/1 015） for Entamoeba coli, 0.4%（4/1 015） for Giardia lamblia, 0.2%（2/1 015） for Iodamoeba butschlii. No infection was found for other parasites. There was no significant sexual difference in the infection rate of Taenia (male 21.2%, 83/391; female 22.0%, 137/624), T. trichiura (male 0; female 0.3%, 2/624), E. vermicularis (male 0.3%, 2/391; female 0.2%, 1/624), E. coli (male 5.4%, 21/391; female 3.4%, 21/624), G. lamblia (male 0.8%, 3/391; female 0.2%, 1/624), or I. butschlii (male 0; female 0.3%, 2/624) (P>0.05). The Taenia sp. infection rate in the age groups of 0-17, 18-29, 30-39, 40-49, 50-59 and ≥ 60 years was 10.9%（18/165）, 14.0%（18/129）, 23.0%（43/187）, 25.3%（61/241）, 26.5%（41/155）, and 28.3%（39/138）, respectively, with significant differences between groups of 0-17 and 40-49 years and between groups of 50-59 and ≥ 60 years (P<0.01). The infection rate of E. coli, T. trichiura and I. butschlii was highest in the age group of 50-59 years（6.5%, 10/155; 0.7%, 1/155; 1.3%, 2/155 respectively）. Among various occupations, farmers showed the highest infection rate for Taenia（44.5%, 147/330）, E. coli（6.4%, 21/33）, E. vermicularis （0.6%, 2/330） and I. butschlii（0.3%, 1/330）; housewives had the highest infection rate for T. trichiura（2.5%, 1/40）; and the students had the highest infection rate for G. lamblia (0.7%, 1/134). Conclusion: There are infections with various intestinal parasites in the participants of Lhasa City, among which taenia infection reaches 20.7% while others have a low level of infection.

siRNA against KIR3DL1 as a potential gene therapeutic agent in controlling HIV-1 infection.

OBJECTIVES: The aim of this study was to develop a small interfering RNA (siRNA) against the expression of KIR3DL1 receptor on natural killer (NK) cells, in order to promote the ability of NK cells to destroy human immunodeficiency virus (HIV)-infected cells and thus prevent failure of siRNA therapy targeting human immunodeficiency virus type 1 (HIV-1) virus among HIV-1 infected patients in vitro. METHODS: A siRNA targeting KIR3DL1 was synthesized and then modified with cholesterol, methylene, and sulfate. The inhibitory action of the siRNAs on primary cultured NK cells was detected. The amount of IFN-γ and TNF-α secretions in NK cells was measured. The intended functions of NK cells in vitro were analyzed by CFSE and PI methods. RESULTS: There were no significant differences in inhibiting the expression of KIR3DL1 on NK cells between the modified and unmodified siRNAs, while inhibition by each of them differed significantly from controls. The amount of IFN-γ and TNF-α secretions in the NK cells was abundant due to unsuccessful expression of KIR3DL1 on NK cells, which further promoted function of the NK cells. CONCLUSION: The siRNA against KIR3DL1 could enhance the ability of the NK cells to kill the HIV-1 infected cells in vitro and successfully prevented the failure of siRNA therapy targeting the HIV-1 virus. Therefore, it can act as a potential gene therapeutic agent among HIV-1 infected people.

Mycoplasmas infection in male HIV/AIDS patients in Jiangsu, China.

Mycoplasmas are widely distributed among animals, plants, and human. The four species namely, Mycoplasmas genitalium(Mg), Mycoplasmas fermentans(Mf), Mycoplasmas pentrans(Mpe), Mycoplasmas pirum(Mpi) are also called AIDS-associated mycoplasmas due to their involvement in the development and outcome of AIDS. To investigate the infection prevalence of Mg, Mf, Mpe and Mpi among male HIV/AIDS patients in Jiangsu Province and to analyze the relationship between pathogenic mycoplasmas and cellular immune function of them. First void urine and venous blood samples were collected and epidemiology questionnaires were administered after informed consent. Nested PCR was performed to determine the infection of Mg, Mf, Mpe and Mpi while ELISA assay was applied to detect interleukin-2(IL-2), interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α). SAS 9.0 software was applied to analyze the data. A total of 713 HIV/AIDS patients were recruited in this study. The overall infection rates of Mg, Mf, Mpe and Mpi are 27.9%, 9.7%, 1.0% and 18.4% respectively. Generally, the infection rates of Mg(χ(2) = 10.311, P = 0.006) and Mpi were declined as the CD4+ cell counts increased, while Mf infection was higher in CD4+ T cell>350/µl group. The levels of cytokines are different with the variance of mycoplasmas infection. Mycoplasma infection among male HIV/AIDS patients is associated with changes in cellular immune response (cytokines). However, the affect of mycoplasmas on the immune function is complex, further studies are still required to elucidate whether mycoplasmas interact with HIV by interfering host immune system.

[Acupuncture and moxibustion in Tunisia].

The development status of acupuncture and moxibustion in Tunisia is introduced in this article. Although acupuncture and moxibustion only has a history of more than 30 years in Tunisia, it is very popular among the local people. Until now, there is one acupuncture and moxibustion center aided and built with the help of the Chinese government. Acupuncture and moxibustion clinical department has been set in some of the hospitals, and acupuncture and moxibustion clinical practice is also carried out in some private clinics. Cost of acupuncture and moxibustion in public hospitals has already been covered by medical insurance. As for education of acupuncture and moxibustion, training courses were set up in medical colleges of Tunisia by Tunisian government which is lectured by Chinese acupuncture experts. Acupuncture and moxibustion has been used to treat many diseases in Tunisia and is warmly welcomed by Tunisian.

[The application of network scale-up method on female sex workers and clients size estimation in Taizhou city].

OBJECTIVE: To estimate the size of female sex workers and clients in Taizhou city. METHODS: A household survey using network scale-up method (NSUM) was conducted among the 3000 community residents in Taizhou city from August to October in 2011, which aimed to estimate the social network size (c value) of Taizhou residents, and the c value was adjusted by demographic characteristics, back estimation and outlier elimination. Using the adjusted c value, the number of acquaintance of female sex workers or clients and the respect level toward female sex workers or clients were used to estimate the size of female sex workers and clients. RESULTS: A total of 2783 valid questionnaires were collected, among which 1380 (49.6%) were collected from Taixing city, 1403 (50.4%) were collected from Jingjiang city. 1334 respondents were male (47.9%) and 1449 (47.9%) respondents were female. The mean age was (39.4 ± 10.7) years. The average personal social network size using original data for Taizhou residents was 525, which differed from place, sex, age, educational level and marriage status. Using the remaining known populations through back estimation, the social network size was 419 and became 424 after the elimination of outliers. The estimated population size for female sex worker was 6370 (95%CI: 5886 - 6853), which accounted for 0.52% (6370/1 229 980) of the total number of female aged from 15 to 49. The estimated population size for clients was 15 202 (95%CI: 14 560 - 15 847), which accounted for 1.28% (15 202/1 190 340) of the total number of males aged from 15 to 49 and the ration of clients to female sex worker was 2.39:1. CONCLUSION: NSUM is an easy and quick way to estimate the size of female sex workers or clients, but the estimated sizes are subject to bias and error due to estimate effect and sample representativeness.

[Analysis on Mycoplasma pirum infection in male HIV/AIDS patients and related 16S rRNA genes in Jiangsu province].

OBJECTIVE: To investigate the prevalence of Mycoplasma pirum (Mpi) in male HIV infected patients, and to identify the 16S rRNA gene of Mpi. METHODS: The first void urine of male HIV/AIDS patients in Jiangsu province was collected for Mpi detection. Purified 16S rRNA gene PCR production was sequenced for analysis on its identification, homogeneity and phylogenetic tree. P1 protein sequence of Mpi was analyzed by Vector NTI Advance 11.0 to calculate the coded amino acid sequence. Homogeneity analysis was conducted between the theoretical amino acid sequence of Mpi and other Mycoplasmas. RESULTS: The prevalence of Mpi in male HIV/AIDS patients was 21.5% while the Mpi prevalence rates in different age groups were significantly different (χ² Mpi = 124.63, P < 0.01). The homogeneity of 18 strains of Mpi was higher than 90%. CONCLUSION: The Mpi prevalence seemed much higher than the results from previous detection on HIV/AIDS patients, suggesting that more attention should be paid on AIDS treatment. More bioinformatic research on gene/nucleotide sequence analysis and forecast should be carried out to identify the molecular characteristics of Mpi.

Emergence of peripheral CD3+CD56+ cytokine-induced killer cell in HIV-1-infected Chinese children.

Cytokine-induced killer (CIK) cells are immune effector cells characterized by co-expression of CD3 and CD56 molecules. We examined the quantities of CIK cells and the changes of these cell expressing NK cell receptors in HIV-1-positive children infected via mother-to-child transmission. The percentage of CIK cells was quantified and the changes in the surface cell receptor profiles in 18 HIV-1-infected children were examined. We found that CIK cell percentages were dramatically increased in HIV-1-infected children. Furthermore, the expressions of CD16, NKp30, NKp44, NKp46, NKp80 and CD244 on CIK cells were decreased, while the expressions of KIR3DL1 and NKG2D on CIK cells were increased in HIV-1-infected children. However, the expressions of KIR2D and NTB-A on CIK cells did not change in the HIV-1-infected children. CIK cells possessed the characteristics of promoting the maturation of dendritic cells and killing functions in HIV-1-infected children. Moreover, serum concentrations of IL-4 and IFN-γ were significantly increased in HIV-1-infected children compared with the HIV-negative controls. These changes likely occurred as a protective mechanism against transmission of maternal HIV-1 virus and thereby helped to limit viral spread, eliminate infected cells and help HIV-1-infected patients to slow the progression to AIDS.

[Observation on relations between onset seasons of peripheral facial paralysis and therapeutic courses and effect of acupuncture].

OBJECTIVE: To observe the influences of seasonal factors on peripheral facial paralysis by acupuncture. METHODS: Four hundred cases of facial paralysis were divided into spring, summer, autumn and winter groups, 100 cases in each group. All these cases were treated by routine puncture. Fengchi (GB 20), Yifeng (TE 17), Qianzheng (Extra), Jiache (ST 6), and Dicang (ST 4), etc. were applied at affect side, once a day. 2 months observation was carried on to compare the clinical therapeutic effects and average courses. The facial symptoms, physical sign and functional activities were taken as observation indexes of therapeutic effect. RESULTS: The effect rate was 78.0% (78/100) in spring group, 82.0% (82/100) in summer group, 89.0% (89/100) in autumn group and 92.0% (92/100) in winter group; the effect rate in summer or autumn group was superior to those in spring group and in summer group (all P < 0.05); the average course was (47.6 +/- 22.3) days in spring group, (43.7 +/- 18.4) days in summer group, (31.5 +/- 11.3) days in autumn group and (22.6 +/-9.2) days in winter group, indicating the significant differences between groups except that between spring and summer group (all P < 0.01). The cured and markedly effective rate was 80.1% (161/201) for wind cold type, 53.5% (61/114) for wind heat type, and 36.5% (31/85) for damp heat type, indicating that it of wind cold type was superior to that of wind heat type or damp heat type (P < 0.001, P < 0.05). CONCLUSION: The syndrome distribution and courses of peripheral facial paralysis are different in different seasons, hence, the diseases should be treated according to attack time and syndromes.

Differences in natural killer cell quantification and receptor profile expression in HIV-1 infected Chinese children.

Natural killer (NK) cells are believed to play a role in the progression of human immunodeficiency virus 1 (HIV-1) disease, and NK cell levels are reduced in individuals with chronic HIV-1 infection. To assess the effects on quantity of NK cells and the changes of NK cell receptors in HIV-1 infected children via mother-to-child transmission, the percentage of NK cells is quantified and the changes in the NK cell receptor profiles in 20 HIV-1 infected children who are not progressing into AIDS were examined. The results showed that NK cell percentage was decreased in the HIV-1 infected children. The expression of NKp30 on NK cells was increased, while the expressions of CD16, NKp44, NKp46, NKp80, NTB-A, CD244, KIR2D, KIR3DL1 and NKG2D on NK cells were decreased in the HIV-1 infected children. NK cell cytolytic activity was elevated in HIV-1 infected children. These results indicate that the acute changes in NK cell percentage and NK cell receptors in HIV-1 infected children are different from the HIV-1 infected adult individuals. Moreover, serum concentrations of IL-18 were elevated in HIV-infected children compared to HIV-uninfected controls. These differences probably play a role in protecting against transmission of maternal HIV-1 virus and guiding the therapeutic strategies for HIV-1 infected children.

[Risk factors which were associated with heroin use during the methadone maintenance treatment among 1301 patients in 9 cities of China.].

OBJECTIVE: To determine the proportion of heroin use among patients who were involved in community-based methadone maintenance treatment (MMT) program and to identify the risk factors associated with heroin use. METHODS: This study was conducted in 9 MMT clinics within 3 provinces. Thirteen hundred and one patients who met the study criteria were selected from each of the five groups with different dosages of methadone users. An administrative questionnaire was applied to explore the demographics, drug abuse-related behaviors and MMT services received by the clients, etc. The prevalence of depression and anxiety among the clients were also collected by SAS and SDS. Urine samples were collected as a biological marker to indicate if heroin had been used. RESULTS: Of the 1301 patients, 76.2% were males. The mean age was (34.6 +/- 6.5) years while 71.7% had an education level of primary school or below. The average daily dosage of methadone was (48.1 +/- 29.4) mg and self-satisfied evaluation score on treatment was 8.6. On average, 27.7% urine samples showed positive opiate evidence. Marital status, employment status, treatment retention, self-satisfied evaluation score on dosage and dropout history were found to be significantly associated with heroin use, while gender, education level and dosage had no significant association with heroin use. It seemed that risk factors that associated with heroin use were different from areas to areas. CONCLUSION: High quality MMT clinic services, high self-satisfied score, longer treatment retention and low dropout rate seemed to have the effects of reducing the risk of ongoing heroin abuse under the methadone maintenance treatment program.

Changes in NK cell counts and receptor expressions and emergence of CD3(dim)/CD56+ cells in HIV-1 infected patients in China.

Natural killer (NK) cells are believed to play a role in human immunodeficiency virus 1 (HIV-1) disease progression, and NK cell levels are reduced in individuals with chronic HIV-1 infection. In the present study, we compared the frequency and phenotype of peripheral blood CD3-CD56+ NK cells in HIV-1 infected patients in China who were infected through different routes of transmission, including heterosexual and homosexual sexual contact, and blood transmission through injection drug use or importation of blood or blood products. The results showed significantly reduced numbers of CD3-CD56+ NK cells with no association with route of transmission. The expression of CD16 on CD3-CD56+ NK cells in HIV-1 infected patients was similar to that in healthy controls. Among the examined receptor (KIR3DL1, NKp80, NKp44, CD244, NKG2D, and NTBA) expressions, only KIR3DL1 and NKp80 expressions on CD3-CD56+ NK cells were suppressed in HIV-1-infected patients compared to healthy controls, and no significant difference was observed between patients upon comparison of different routes of transmission. A subset of CD3(dim)/CD56+ cells was dramatically increased in HIV-1-infected patients. This study suggests that changes in NK cell count and receptors are not related to the route of HIV-1 transmission. A new subset of CD3(dim)/CD56+ cells emerged only in HIV-1-infected patients, and may play a role in limiting viral spread, eliminating infected cells, and slowing the progression from HIV-1 infection to AIDS.

[Study on fluoroquinolone resistance and the relationship between resistance and mutations of gyrA and parC in Neisseria gonorrhoeae].

OBJECTIVE: To study the phenotypic and genotypic resistance to Fluoroquinolones in Neisseria gonorrhoeae (NG) isolated in Jiangsu province of China. METHODS: In-vitro, susceptibility testing of ciprofloxacin and levofloxacin against ninety-five clinical isolates were determined by agar dilution method. Detection of mutation in the gyrA and parC genes was performed by polymerase chain reaction (PCR) assay and sequence analysis. RESULTS: The clinical isolates demonstrated 100% resistance to ciprofloxacin. Based on gyrA and parC mutations, 18 types could be categorized among the 54 isolates. Based on the same gyrA mutations,isolates with high MIC appeared to have had more mutations in parC gene. CONCLUSION: The status of resistance to ciprofloxacin in NG was quite serious, and ciprofloxacin treatment for the treatment of NG infections in Jiangsu province should not be recommended. The results from this study suggested that mutations in the parC gene had contributed to the development of high Fluoroquinolone resistance in NG.

Antimicrobial susceptibility of Neisseria gonorrhoeae isolated in Jiangsu Province, China, with a focus on fluoroquinolone resistance.

In this study, the phenotypic and genotypic resistance to fluoroquinolones in Neisseria gonorrhoeae isolated in Jiangsu Province, China, was analysed. In vitro susceptibility testing of eight antimicrobial agents, including ciprofloxacin and levofloxacin, against 95 clinical isolates was carried out. Detection of mutations in the gyrA and parC genes was performed by sequence analysis. The clinical isolates demonstrated 100% resistance to ciprofloxacin and 98.9% non-susceptibility to levofloxacin. All of the isolates were susceptible to cefotaxime and ceftriaxone. For cefepime, spectinomycin and tetracycline, 98.9, 94.7 and 1.1% of the isolates were susceptible, respectively. None of the isolates was susceptible to penicillin. Five types based on gyrA mutations could be categorized among 54 isolates with seven different mutation sites found on their parC gene. Analysis of sequence results showed that the gyrA mutation Asp-95-->Ala and the parC mutations Ser-87-->Arg and Ser-87-->Asn made a significant contribution to the resistance to fluoroquinolones, in addition to double mutations found in each gene. Therefore, the use of fluoroquinolones in the treatment of N. gonorrhoeae infections in Jiangsu Province is not recommended, while the use of third- and fourth-generation cephalosporins and spectinomycin is recommended.

[Sequential analysis of detection Mycoplasma in different positions of female genital tract].

OBJECTIVE: To discuss the main parasitic position of Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) in female genital tract. METHODS: Using the standard aseptic cotton swab to collect secretion in vaginal fornix and orificium internum uteri, to culture Uu and Mh in Mycoplasma ID medium of France Bio-Merieux Co. According to double-direction quality reaction sequential test design, detection results of different position were analyzed. RESULTS: Total positive and > or = 10(4) ccu/ml positive of Uu in vaginal fornix were significantly higher than that in orificium internum uteri. Total positive of Mh in vaginal fornix was significantly higher than that in orificium internum uteri as well. CONCLUSION: In order to raise the detectable rate of Mycoplasma, we suggested that the secretion in vaginal fornix position be collected.