Analysis of genetic diversity among Chinese Cyclocybe chaxingu strains using ISSR and SRAP markers.

Background: Cyclocybe chaxingu is an edible and medicinal fungal species commonly cultivated in China. The major problems currently facing by growers of C. chaxingu is the random labeling of strains and strains aging and degeneration. Therefore, an evaluation of genetic diversity is essential for the conservation and reproducing programs of this species. Methods: In the present study, 24 widely cultivated strains were collected from the main producing areas of China, and the genetic diversity analysis was performed. DNA polymorphism among these Chinese C. chaxingu strains was analyzed using inter-simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers. Results: Eight ISSR primers amplified a total of 75 DNA fragments of which 61 (81.33%) were polymorphic. Fifteen SRAP primer combinations amplified 166 fragments of which 132 (79.52%) were polymorphic. Cluster analysis showed that the C. chaxnigu strains fall into five groups with a genetic distance values ranging from 0.06 to 0.60 by ISSR analysis, while the SRAP analysis divided the test strains into four groups within the range of genetic distance from 0.03 to 0.57. The results of the present study reveal a high level of genetic diversity among the widely cultivated C. chaxingu strains.

Retiboletus atrofuscus (Boletaceae, Boletales), a new species from China.

Based on morphological and phylogenetic analyses, an ectomycorrhizal fungus collected from Jiangxi, China, is described as a new species. R. atrofuscus is morphological characterized by a dark gray to black pileus, white to pale yellowish white hymenophore, a grayish brown to pale yellow context, which turning orange-yellow to rusty yellow when injured, a prominently and coarsely white to pale yellow to black reticulation on upper 3/4 or entire stipe, grayish yellow to pale yellow to yellowish brown basidiospores in KOH with measuring (7-)7.5-13(-13.5) × (3-)3.5-5(-5.5) µm. Phylogenetic analysis results showed that R. atrofuscus formed an independent lineage within Retiboletus. Descriptions and hand drawings of the new species and comparisons with similar species are presented.

Effects of Chronic Hypoxic Environment on Cognitive Function and Neuroimaging Measures in a High-Altitude Population.

Objective: This study aimed to investigate the effects of long-term hypoxic environment exposure on cognitive ability and neuroimaging characteristics in a highland population in China. Methods: Health system workers in Maduo County (4,300 m above sea level) and Minhe County (1,700 m above sea level) were selected as research participants and divided into a high-altitude (HA) group and low-altitude (LA) group, respectively. Cognitive ability was assessed using the Montreal Cognitive Assessment (MoCA), Verbal Fluency Test (VFT), Symbol Digit Modalities Test (SDMT), Trail Making Test A and B (TMT), Digit Span Test (DST), and Rey Auditory Verbal Learning Test (RAVLT). All participants underwent a magnetic resonance imaging (MRI) scan, resting state functional MRI scan, and diffusion tensor imaging to clarify changes in regional gray matter (GM) volume, anisotropy index (FA), local consistency (ReHo), and low-frequency oscillation amplitude (ALFF). Results: The HA group had significantly lower MoCA, DST, VFT, RAVLT, and TMT scores compared to the control group. No significant differences were found in SDMT score. Furthermore, compared to the LA group, the HA group had significantly lower GM density of the left olfactory cortex, right medial orbital superior frontal gyrus, bilateral insula, left globus pallidus, and temporal lobe (left superior temporal gyrus temporal pole, bilateral middle temporal gyrus temporal pole, and right middle temporal gyrus). In terms of FA, compared with the LA group, the HA group had lower values for the corpus callosum, corpus callosum knee, bilateral radiative corona, and left internal capsule. The HA group had lower ALFF values of the left cerebellum, left putamen, left orbital inferior frontal gyrus, and left precuneus, but higher ALFF values of the left fusiform gyrus, bilateral inferior temporal gyrus, left orbital superior frontal gyrus and medial superior frontal gyrus, compared to the LA group. There was no significant group difference in ReHo values. Conclusion: Our findings suggest that a chronic hypoxic environment can induce extensive cognitive impairment. Decreased GM density in multiple brain regions, damaged nerve fibers, and unbalanced neuronal activity intensity in different brain regions may be the structural and functional basis of cognitive impairment due to hypoxia.

[The level of the stress biomarker salivary alpha amylase is correlated with semen quality in infertile young men].

Objective: To investigate the relationship between the level of the stress biomarker salivary alpha amylase (SAA) and semen quality in infertile young men. METHODS: Totally, 313 infertile and 96 normal healthy men, aged 20－40 years old, were enrolled in this study. The SAA levels and semen parameters of the subjects were measured and compared between the two groups. RESULTS: Compared with the normal healthy controls, the young infertility patients showed a significantly higher SAA level (ï¼»141.04 ± 44.13ï¼½ vs ï¼»151.48 ± 38.42ï¼½ µmol/L, P < 0.05) and percentage of immotile sperm (IMS) (ï¼»39.98 ± 14.53ï¼½% vs ï¼»64.48 ± 26.32ï¼½%, P < 0.05), but lower sperm concentration (ï¼»44.23 ± 21.63ï¼½ vs ï¼»32.42 ± 23.07ï¼½ ×106/ml, P < 0.05) and percentage of progressively motile sperm (PMS) (ï¼»52.13 ± 15.42ï¼½% vs ï¼»27.91 ± 21.22ï¼½%, P < 0.05). Sperm concentration (ï¼»26.33 ± 31.83ï¼½ vs ï¼»35.28 ± 27.70ï¼½ ×106/ml, P < 0.05) and the percentage of PMS were remarkably lower in the infertile men with a high than in those with a low SAA level (ï¼»19.85 ± 21.55ï¼½% vs ï¼»31.70 ± 20.02ï¼½%, P < 0.05), while the percentage of IMS was higher in the former than in the latter group (ï¼»74.19 ± 26.84ï¼½% vs ï¼»59.92 ± 24.85ï¼½%, P < 0.05). The SAA level in the young infertility patients was correlated positively with the percentage of IMS (r = 0.170, P < 0.01), but negatively with sperm concentration (r = －0.227, P < 0.01) and the percentage of PMS (r = －0.468, P < 0.01). CONCLUSIONS: The stress biomarker salivary alpha amylase level in infertile young men is negatively correlated with semen quality, and therefore semen parameters can be improved by reducing the stress level.

Neuroprotective effects of protocatechuic aldehyde against neurotoxin-induced cellular and animal models of Parkinson's disease.

Protocatechuic aldehyde (PAL) has been reported to bind to DJ-1, a key protein involved in Parkinson's disease (PD), and exerts potential neuroprotective effects via DJ-1 in SH-SY5Y cells. In this study, we investigated the neuroprotective pharmacological effects of PAL against neurotoxin-induced cell and animal models of PD. In cellular models of PD, PAL markedly increased cell viability rates, mitochondrial oxidation-reduction activity and mitochondrial membrane potential, and reduced intracellular ROS levels to prevent neurotoxicity in PC12 cells. In animal models of PD, PAL reduced the apomorphine injection, caused turning in 6-OHDA treated rats, and increased the motor coordination and stride decreases in MPTP treated mice. Meanwhile, in an MPTP mouse model, PAL prevented a decrease of the contents of dopamine (DA) and its metabolites in the striatum and TH-positive dopaminergic neuron loss in the substantia nigra (SN). In addition, PAL increased the protein expression of DJ-1 and reduced the level of α-synuclein in the SN of MPTP lesioned mice. PAL also increased the spine density in hippocampal CA1 neurons. The current study demonstrates that PAL can efficiently protect dopaminergic neurons against neurotoxin injury in vitro and in vivo, and that the potential mechanisms may be related to its effects in increasing DJ-1, decreasing α-synuclein and its growth-promoting effect on spine density.

Seasonal variation in semen quality in China.

In this study, we assessed the effect of seasonal variation on the semen quality in Chinese by using the average highest temperature (AHT) of the ejaculation day for partitioning season periods. A total of 13 635 semen samples were collected and analysed according to the AHT of the ejaculation day. Semen volume, sperm concentration and the percentage of spermatozoa with normal morphology in midsummer (AHT > 30 °C) were significantly lower than those in other periods of the year. Sperm motility with AHT 10-20 °C in winter and spring was significantly higher than that in other seasons. The percentage of head defects spermatozoa with AHT < 10 °C in winter was significantly lower than that in other periods. We observed that there was a significant effect of season on the semen parameters. Highest environment temperature of the day may be a determining factor for the seasonal variations in semen quality.

Evaluation of indices in differentiation between iron deficiency anemia and beta-thalassemia trait for Chinese children.

SUMMARY: Iron deficiency anemia (IDA) and beta-thalassemia trait (beta-TT) are the most common microcytic anemia. Kinds of discrimination indices have been established as simple, fast, and inexpensive means since 1970s. This study evaluates the validity of 12 indices in differentiation between IDA and beta-TT for Chinese children. The 12 discrimination indices were red blood cell (RBC) count, RBC distribution width, Mentzer Index, Green and King Index, RBC distribution width Index, England and Fraser Index, Shine and Lal Index, Srivastava Index, Ricerca Index, Ehsani Index, Sirdah Index, and Huber-Herklotz-Index. A total of 300 children (beta-TT, n=127 and IDA, n=173) were enrolled in the study. We calculated the sensitivity and specificity, positive and negative predictive values, and Youden's index for each index. We also established new cut-off values by receiver operating characteristic curves. Green and King Index showed the highest reliability, as it had the largest area under the curve (AUC=0.947), whereas Srivastava Index and Shine and Lal Index showed the lowest reliability (AUC=0.679 and 0.532, respectively). We concluded Green and King Index, Ricerca Index, and England and Fraser Index could be used in discrimination between IDA and beta-TT in Chinese children.

[Surgical treatment of ankle fracture in the elderly].

OBJECTIVE: To analysis the outcomes and the technique of surgical operation of osteoporotic ankle fracture in the elderly. METHODS: One hundred and twenty-eight patients with osteoporotic ankle fracture were treated by surgical operation from January 2003 to December 2008, included 26 males and 102 females, with an average age of 69 years old ranging from 50 to 82 years. According to Lange-Hanson classification, there were 113 cases of supinatio-extorsion, 18 of pronate-extorsion, 5 of supinatio-adduction, 2 of pronate-abduction. The methods of fixation, the complications, and the AOFAS scores after operation were recorded. RESULTS: All the 128 patients were followed up from 6 months to 6 years (averaged in 36 months). Delayed wound healing occurred in 11 cases (8.6%). There was no deep infection, nonunion of the fracture, and revision. The average AOFAS scores after operation was 83.6 +/- 6.3 (the mean pain scores were 33.5 +/- 5.6, the mean function scores were 40.2 +/- 7.4, the mean alignment scores were 10). CONCLUSION: Surgical treatment should be the first choice in osteoporotic ankle fracture in most cases. Improved procedures can make the results better.

Environmental lead exposure among children in Chengdu, China: blood lead levels and major sources.

A survey was performed to know blood lead level (BLL) of children under seven and the risk factors of high BLL in Chengdu, China in 2004. The mean BLL in children under seven in Chengdu was 63.88 microg/L. The detection rate of high BLL was 8.21%. Chengdu is a moderate popular region of lead poisoning. Substitute of breast milk, living at the base floor or in one-storey houses and houses near the streets are the risk factors of high BLL (p < 0.05). The risk of anorexia, spasm and impaired concentration is higher in children whose BLL is higher than those whose BLL is lower (p < 0.05). Living circumstances, feeding patterns, and eating habits affect BLL, which in turn influences children's health status.

Treatment of calcaneal osteomyelitis with free serratus anterior muscle flap transfer.

BACKGROUND: Chronic calcaneal osteomyelitis is a difficult problem to eradicate. The infected bone and soft tissue can rarely be successfully treated conservatively. But after aggressive debridement, there is usually a large defect in the calcaneus and soft tissue. Free serratus anterior muscle flap can not only fill the defect but can help eliminate the infection of the bone and soft tissue. MATERIALS AND METHODS: Thirteen patients with calcaneal osteomyelitis were treated with free serratus anterior muscle flap transfer and skin graft from 2000 to 2006. The mean patient age was 36 (range, 18 to 48) years. Four patients were women and nine were men. The followup averaged 34 months (range, 6 months to 6 years). RESULTS: All flaps survived and there was no infection recurrence. The mean operation time was 6.3 +/- 1.3 hours. Partial skin graft necrosis was found in one patient, which resolved uneventfully after another skin graft. There was minimal donor site morbidity. Winged scapula or dysfunction of the shoulder was not found in any of the patients. CONCLUSION: The serratus anterior muscle has many advantages owing to its anatomical features. We believe it is a good option for treatment of calcaneal osteomyelitis.

Reconstruction of soft tissue defects of the lower limb using the free serratus anterior flap.

OBJECTIVE: To present the application and clinical results of soft tissue reconstruction of the lower limb with the free serratus anterior muscle flap. METHODS: Twenty Chinese adult cadavers were studied to determine detailed anatomical information about the serratus anterior muscle flap. From 1997 to 2007, 82 patients with soft tissue defects of the lower limbs were treated with free serratus anterior muscle flaps and skin grafts. There were 24 females and 58 males, aged from 22 to 63 years (mean 34). The patients were followed up for an average of 30 months (range, 8 months-5 years). RESULTS: All flaps survived except for one, in which necrosis occurred. Six patients developed partial necroses of the muscle flap or skin graft. Five of them healed with debridement or wound care, and one healed with a repeat skin graft. There was no notable donor site morbidity. Dysfunction of the shoulder was not found in any of the patients. CONCLUSION: The serratus anterior muscle flap is a good option for the treatment of soft tissue defects of the lower limb. This muscle flap has many advantages owing to its anatomical features.

Purification and characterization of a barley aleurone abscisic acid-binding protein.

A protein designated ABAP1 and encoded by a novel gene (GenBank accession number AF127388) was purified and shown to specifically bind abscisic acid (ABA). ABAP1 protein is a 472-amino acid polypeptide containing a WW protein interaction domain and is induced by ABA in barley aleurone layers. Polyclonal antiidiotypic antibodies (AB2) cross-reacted with purified ABAP1 and with a corresponding 52-kDa protein associated with membrane fractions of ABA-treated barley aleurones. ABAP1 genes were detected in diverse monocot and dicot species, including wheat, tobacco, alfalfa, garden pea, and oilseed rape. The recombinant ABAP1 protein optimally bound (3)H-(+)-ABA at neutral pH. Denatured ABAP1 protein did not bind (3)H-(+)-ABA, nor did bovine serum albumin. The maximum specific binding as shown by Scatchard plot analysis was 0.8 mol of ABA mol(-1) protein with a linear function of r(2) = 0.94, an indication of one ABA-binding site with a dissociation constant (K(d)) of 28 x 10(-9) m. ABA binding in aleurone plasma membranes showed a maximum binding capacity of 330 nmol of ABA g(-1) protein with a K(d) of 26.5 x 10(-9) m. The similarities in the dissociation constants for ABA binding of the recombinant protein and that of the plasma membranes suggest that the protein within the plasma membrane fraction is the native form of ABAP1. The stereospecificity of ABAP1 was established by the incapability of ABA analogs and metabolites, including (-)-ABA, trans-ABA, phaseic acid, dihydrophaseic acid, and (+)-abscisic acid-glucose ester, to displace (3)H-(+)-ABA bound to ABAP1. However, two ABA precursors, (+)-ABA aldehyde and (+)-ABA alcohol, were able to displace (3)H-(+)-ABA, an indication that the structural requirement of ABAP1 at the C-1 position is not strict. Our data show that ABAP1 exerts high binding affinity for ABA. The interaction is reversible, follows saturation kinetics, and has stereospecificity, thus meeting the criteria for an ABA-binding protein.

A comparative analysis of two cDNA clones of the cellulase gene family from anaerobic fungus Piromyces rhizinflata.

Cellulase family and some other glycosyl hydrolases of anaerobic fungi inhabiting the digestive tract of ruminants are believed to form an enzyme complex called cellulosome. Study of the individual component of cellulosome may shed light on understanding the organization of this complex and its functional mechanism. We have analysed the primary sequences of two cellulase clones, cel5B and cel6A, isolated from the cDNA library of ruminal fungus, Piromyces rhizinflata strain 2301. The deduced amino acid sequences of the catalytic domain of Cel5B, encoded by cel5B, showed homology with the subfamily 4 of the family 5 (subfamily 5(4)) of glycosyl hydrolases, while cel6A encoded Cel6A belonged to family 6 of glycosyl hydrolases. Phylogenetic tree analysis suggested that the genes of subfamily 5(4) glycosyl hydrolases of P. rhizinflata might have been acquired from rumen bacteria. Cel5B and Cel6A were modular enzymes consisting of a catalytic domain and dockerin domain(s), but not a cellulose binding domain. The occurrence of dockerin domains indicated that both enzymes were cellulosome components. The catalytic domain of the Cel5B (Cel5B') and Cel6A (Cel6A') recombinant proteins were purified. The optimal activity conditions with carboxymethyl cellulose (CMC) as the substrate were pH 6.0 and 50 degrees C for Cel5B', and pH 6.0 and 37-45 degrees C for Cel6A'. Both Cel5B' and Cel6A' exhibited activity against CMC, barley beta-glucan, Lichenan, and oat spelt xylan. Cel5B' could also hydrolyse p-nitrophenyl-beta-d-cellobioside, Avicel and filter paper while Cel6A' did not show any activity on these substrates. It is apparent that Cel6A' acted as an endoglucanase and Cel5B' possessed both endoglucanase and exoglucanase activities. No synergic effect was observed for these recombinant enzymes in vitro on Avicel and CMC.

A variant of Orpinomyces joyonii 1,3-1,4-beta-glucanase with increased thermal stability obtained by random mutagenesis and screening.

A thermostable variant of an Orpinomyces joyonii beta-glucanase was identified by screening a mutant library constructed using error-prone PCR products. The mutant, designated 2011D, had one amino acid substitution (Val replaced Asp-70). 2011D showed similar catalytic efficiency to its wild-type enzyme, LicA. The temperature at which 50% inactivation occurred after heat treatment for 10 min was increased by 14 degrees C for 2011D, in comparison to those of wild-type enzyme.