There are few data on the molecular epidemiology of cryptococcosis in China. Here we investigated the species distribution, molecular types and antifungal susceptibilities of 312 Cryptococcus neoformans species complex isolates from ten hospitals over 5 years. Isolates were identified by internal transcribed spacer (ITS) sequencing and by two matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems. Multilocus sequence typing (MLST) was used to verify species/variety and to designate molecular types. Susceptibility to six antifungal drugs was determined by the Sensititre YeastOne™ method. Cryptococcus neoformans was the predominant species (305/312 isolates (97.8%), all were ITS type 1, serotype A), of which 89.2% (272/305) were C. neoformans var. grubii MLST sequence type (ST) 5 and 6.2% (19/305) were ST31. Other C. neoformans var. grubii STs were rare but included six novel STs. Only two strains were C. neoformans var. neoformans (both serotype AD). Cryptococcus gattii was uncommon (n = 7, four ITS types) and comprised five MLST STs including one novel ST. For C. neoformans var. grubii, the proportion of isolates with non-wild-type MICs to fluconazole significantly rose in the fourth study year (from 0% (0/56 isolates) in the first year to 23.9% (17/71) in the fourth year), including five isolates with fluconazole MICs of ≥32 mg/L. The study has provided useful data on the species epidemiology and their genetic diversity and antifungal susceptibility. The proportional increase in isolates with non-wild-type MICs to fluconazole is noted.

PCR ribotyping is the most commonly used Clostridium difficile genotyping method, but its utility is limited by lack of standardization. In this study, we analyzed four published whole genomes and tested an international collection of 21 well-characterized C. difficile ribotype 027 isolates as the basis for comparison of two capillary gel electrophoresis (CGE)-based ribotyping methods. There were unexpected differences between the 16S-23S rRNA intergenic spacer region (ISR) allelic profiles of the four ribotype 027 genomes, but six bands were identified in all four and a seventh in three genomes. All seven bands and another, not identified in any of the whole genomes, were found in all 21 isolates. We compared sequencer-based CGE (SCGE) with three different primer pairs to the Qiagen QIAxcel CGE (QCGE) platform. Deviations from individual reference/consensus band sizes were smaller for SCGE (0 to 0.2 bp) than for QCGE (4.2 to 9.5 bp). Compared with QCGE, SCGE more readily distinguished bands of similar length (more discriminatory), detected bands of larger size and lower intensity (more sensitive), and assigned band sizes more accurately and reproducibly, making it more suitable for standardization. Specifically, QCGE failed to identify the largest ISR amplicon. Based on several criteria, we recommend the primer set 16S-USA/23S-USA for use in a proposed standard SCGE method. Similar differences between SCGE and QCGE were found on testing of 14 isolates of four other C. difficile ribotypes. Based on our results, ISR profiles based on accurate sequencer-based band lengths would be preferable to agarose gel-based banding patterns for the assignment of ribotypes.

Macrolide resistance in

Sinopodophyllum hexandrum (Royle) Ying is an important medicinal and endangered species. Inter-simple sequence repeats (ISSR) analysis was conducted on seven natural populations from western SichuanProvince to investigate the genetic diversity of S. hexandrum. Leaf samples of 140 individuals were collected. Of the 139 discernible fragments generated by 12 selected primers (among 100 primers), 54 appeared to be polymorphic. The percentage of polymorphic bands (PPB) was 38.85% at the species level, and PPB within a population ranged from 7.91% to 23.74%. Low levels of genetic variation (He = 0.092, Ho = 0.142) and high levels of genetic differentiation among the populations (G st = 62.25%) was detected on the basis of results from POPGENE and analysis of molecular variance (AMOVA), respectively. Furthermore, the limited gene flow (Nm = 0.361) may result from biological characteristics, such as self-pollination and short distance seed dispersal. Based on the genetic and ecological information available for S. hexandrum, we propose some appropriate strategies for the conservation of the endangered medicinal species in this region, namely rescuing and conserving the core populations for in situ conservation and sampling and preserving more populations with fewer individuals from each population for ex situ conservation. (2006). ISSR analysis of the genetic diversity of the endangered species Sinopodophyllym hexandrum (Royle) Ying from western Sichuan Province, China.

Purpose: Hypertension is a major risk factor for cardiovascular diseases and stroke, and it requires lifelong medication. This study aimed to investigate the factors impacting on Health-Related Quality of Life (HRQoL) among hypertensive patients in Chongqing, China, and to provide evidence-based strategies to improve their HRQoL. Methods: This cross-sectional survey was conducted in Chongqing, China. Of 600 randomly selected patients, 586 patients agreed to participate and 567 patients completed the survey. A SF-36 (Medical Outcomes Study (MOS) Short Form Health Survey questionnaire) that included eight domains: physical functioning, role limitations due to physical problems, body pain, general health, vitality, social function, role limitations due to emotional problems, and mental health was used to measure HRQoL. Linear regressions were used; each domain of HRQoL was measured in the stratification of sex. Results: Self-perceived relatively low economic burden caused by hypertension and regular physical activity had a positive impact on HRQoL (p < 0.05) for both men and women. For women, younger age was associated with higher scores of measuring physical functioning and body pain. Living with more than three family members had a positive impact on domains, including physical functioning. Emotional self-regulation had a positive association with women’s mental health. Alcohol use for men was associated with higher scores in physical and mental health measures, and emotional self-regulation showed some positive impact on general health. Conclusion: Perceived economic burden caused by hypertension was the most common factor impacting on patients’ HRQoL. Female patients were more susceptible when compared to male patients. Health intervention strategies need to be further explored and adapted to the context of improving HRQoL for patients who suffer from hypertension and other chronic non-communicable diseases.

Background Birt-Hogg-Dubé syndrome (BHDS) is an autosomal dominant disease featured by lung cysts, spontaneous pneumothorax, fibrofolliculomas and renal tumors. The causative gene for BHDS is the folliculin (FLCN) gene and more than 200 mutations have been reported in FLCN, mostly truncating mutations. The aim of this study is to better characterize the clinical features and mutation spectrum of Chinese BHDS patients and to systematically evaluate the effects of non-truncating mutations on mRNA splicing pattern. Methods We enrolled 47 patients from 39 unrelated families with symptoms highly suggestive of BHDS after informed consent and detailed clinical data were collected. Exon sequencing followed by multiplex ligation-dependent probe amplification testing were applied for mutation screening. The effects of non-truncating mutations, including 15 missense mutations and 6 in-frame deletions, on mRNA splicing were investigated by minigene assays. Results A total of 24 FLCN germline variants were found in 39 patients from 31 distinct families. Out of these patients, 100% (36/36) presented with lung cysts and 58.3% (21/36) had experienced spontaneous pneumothorax. Seventeen mutation carriers had skin lesions (47.2%, 17/36) and 9 (30%, 9/30) had kidney lesions including 8 with renal cysts and 1 with renal hamartoma. Among all detected variants 14 (58.3%, 14/24) were novel, including 11 variants classified to be pathogenic and 3 variants of uncertain significance. None of 21 non-truncating mutations changed the mRNA splicing pattern of minigenes. Conclusions We found different clinical features of Chinese BHDS patients compared with Caucasians, with more lung cysts and pneumothorax but fewer skin lesions and malignant renal cancer. Chinese patients with BHDS also have a different mutation spectrum from other races. Non-truncating mutations in FLCN did not disrupt mRNA splicing pattern, in turn supporting the hypothesis that these mutations impair folliculin function by disrupting the stability of the FLCN gene product.

This study outlined the epidemiologic data of invasive yeast infections and highlighted the need for continuous monitoring of azole resistances among C. glabrata and C. tropicalis isolates in Beijing.

A data analysis of yeast collections from the National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) programme in 2013 revealed a sudden increase in the proportion of Candida parapsilosis complex isolates (n = 98) in one participating hospital (Hospital H). Out of 443 yeast isolates submitted to the CHIF-NET reference laboratory by Hospital H (2010–2014), 212 (47.9%) were identified as C. parapsilosis sensu stricto by sequencing analysis of the internal transcribed spacer region and D1/D2 domain of the 26S rRNA gene. Among the 212 C. parapsilosis sensu stricto isolates, 176 (83.0%) bloodstream-based isolates and 25 isolates from tip cultures of various vascular catheters from 25 patients with candidaemia, were subjected to microsatellite genotyping, and a phylogenetic relationship analysis was performed for 152 isolates. Among the 152 isolates, 45 genotypes (T01 to T45) were identified, and two prevalent genotypes (63.8%) were found: T15 (n = 74, 48.7%) and T16 (n = 23, 15.1%). These two main clones were confined mainly to three different wards of the hospital, and they persisted for 16–25 months and 12–13 months, respectively. The lack of proper coordination between the clinical microbiology laboratory and infection control staff as part of public health control resulted in the failure to timely identify an outbreak, which led to the wide and long-term dissemination of C. parapsilosis sensu stricto in Hospital H.