[Discussion on related problems of model species of fungus： Paecilomyces hepialid].

Paecilomyces hepiali is a new species of fungus isolated from a field collection of Ophiocordyceps sinensis from Baima snow mountain, Diqing Tibetan Autonomous Prefecture, Yunnan Province by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. The specimen was identified and named as Paecilomyces hepiali by Qing-Tao Chen, the professor of the Institute of Microbiology, Chinese Academy of Sciences (Paecilomyces hepiali) (2008), who identified a dried culture of living strain 82-2 as the holotype. Until now, the holotype (the voucher specimen) was deposited in the Herbarium of the Institute of Chinese Materia Medica (HICMM), China Academy of Chinese Medical Sciences, Beijing. The P. hepiali neotype designated by the paper "Neotypification of P. hepiali (Hypocreales)" published in TAXON 64 (1) by Yao Yi-Jian et al. in February 2015 is untenable.

[Cloning and expression analysis of pathogenesis-related protein 1 gene of Panax notoginseng].

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.

[Restriction endonuclease digest - melting curve analysis: a new SNP genotyping and its application in traditional Chinese medicine authentication].

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.

[Mechanism exploration on synthesis of secondary metabolites in Sorbus aucuparia cell cultures treated with yeast extract].

Suspension cultures cell of Sorbus aucuparia (SASC) was used as materials, the changes of physiological and biochemical indexes of SASC after treatment with yeast extract (YE) were detected, and the synthetic mechanism of secondary metabolites in SASC treated with YE was preliminarily explored. The results were as follows: under the assay conditions, SASC was induced to synthesize five biphenyl compounds, and these compounds content changed differently with induction time prolonging; YE treatment inhibited cell growth, the culture medium pH was gradually reduced after treatment; water-soluble protein content showed a trend of slow decline, which was significantly increased in YE treatment group (YE group) compared with the control group (CK group), the maximum relative content was 147.76% in contrast with CK group; both YE group and CK group were extracellular Ca2+ flow influx, but the YE group flow was significantly slow than CK group. The results indicate that YE induced the cells in a stress state, which was not conducive to the growth of cells and forced the cells to synthesize biphenyl compounds against external stress; water-soluble protein may serve as intracellular enzymes involved in the synthesis of compounds regulation; Ca2+ may as signal molecule mediate cell signal transduction respond to YE stress.

[Growth and accumulation of shikonin compounds of two kinds of cells in suspension culture of Arnebia euchroma].

Via studying the phenotype, growth curve and secondary metabolites of two kinds of suspension culture cell of Arnebia euchroma, the kinetics parameters of growth and accumulation of shikonin compounds in cell suspension culture of A. euchroma was obtained through simulating and modeling. This Study found that the red high-yielding one was a fine cell line for producing shikonin compounds, and the white low-yielding one may be a mutant. The first-order and second-order derivative of the fitting function were obtained by fitting the Logistic model of growth curve to get the growth rate and growth acceleration curve of the suspended cells. It is found that the best period to subculture was the 15th day cultured in fresh medium, and the best period of the induction process was the 13th-14th day. When compared the growth rate of the red line and the shikonin compounds accumulation curve, it is found that the rapid growth of the biomass of cells was not conducive to the synthesis and accumulation of shikonin compounds.

[Identification of a novel PHOX2A gene mutation in a Chinese family with congenital fibrosis of extraocular muscles type 2].

OBJECTIVE: To investigate potential mutation of PHOX2A (or ARIX) gene in a Chinese family affected with congenital fibrosis of extraocular muscles tyep 2 (CFEOM2). METHODS: Genomic DNA was obtained from affected and unaffected members of the family. With an ABI PRSIM Linkage Mapping Set-MD10 kit, selected markers flanking the PHOX2A locus were used for linkage analysis. Exons of PHOX2 gene were amplified and sequenced. A total of 100 normal subjects were recruited as controls. RESULTS: Genetic linkage was found at 11q13 between D11S4151 and D11S1320 and the PHOX2A gene. DNA sequencing has identified a heterozygous mutation in the exon 2 of the gene (227T to G, N76K). The same mutation was not found in the unaffected and 100 normal controls. CONCLUSION: A mutation of the PHOX2A gene 227T to G is responsible for the onset of congenital fibrosis of extraocular muscles type 2 in this Chinese family.

[Cloning and bioinformatic analysis of FatB genes in Lonicera japonica Thunb and its substitutes].

A FatB unigene was obtained from the transcriptome dataset of Lonicera japonica Thunb. Full-length FatB cDNA was cloned from buds of Lonicera japonica Thunb., Lonicera japonica Thunb. var. chinensis (Wats.) Bak., Lonicera hypoglauca Miq. and Lonicera dasystyla Rehd. using RT-PCR technology, and named as LJFatB, LHFatB, LJCFatB and LDFatB. The results of bioinformatic analysis showed that LJFatB, LJCFatB, LHFatB and LDFatB and Arabidopsis thaliana AtFatB had a closely relationship. Nucleotide sequences and protein secondary structure of LJFatB, LJCFatB, LHFatB and LDFatB are different and their proteins had conserved FatB substrate binding sites and catalytic activity sites. Transcriptive level of LJFatB, LJCFatB, LHFatB and LDFatB in bud was not significantly different. Therefore, LJFatB, LJCFatB, LHFatB and LDFatB could have the same biological function as AtFatB.

[Authentication of Lonicera japonica using bidirectional PCR amplification of specific alleles].

OBJECTIVE: To identify SNP in flos Lonicerae, and authenticate Lonicera japonica from its adulterants and the mixture by using bidirectional PCR amplification of specific alleles (Bi-PASA). METHOD: SNP of L. japonica and its adulterants was identified by using ClustulW to align trnL-trnF sequences of the Lonicera genus from GenBank database. Bi-PASA primer was designed and the PCR reaction systems including annealing temperature optimized. Optimized result was performed in 84 samples to authenticate L. japonica, its adulterants and the mixture. RESULT: When the annealing temperature was 61 degrees C, DNA from L. japonica would be amplified 468 bp whereas PCR products from all of the 9 adulterants were 324 bp. The established method also can detect 5% of intentional adulteration DNA into L. japonica. CONCLUSION: The Bi-SPASA could authenticate L. japonica from its adulterants and the mixture.

[Effects of plant growth regulator mepiquat chloride on content of active compounds in Scutellaria baicalensis].

OBJECTIVE: To study the effect of plant growth regulator mepiquat chloride (DPC) on the growing development of plant and the content of active compounds in Scutellaria baicalensis. METHOD: After spraying DPC during the seedling period, the length of taproot, fresh weight of root, diameter of taproot and the length of stem were measured. The contents of baicalin, baicalein and wogonin were determined by HPLC, respectively. Total flavonoids and scavenging DPPH were determined with ultraviolet spectrophotometry. RESULT: After spraying DPC, fresh weight of S. baicalensis root was significant increased, and root diameter was also increased. The contents of baicalin and total flavonoids were significantly increased and baicalein and wogonin were decreased remarkably. Compared with controls, scavenging activity of ethanol extracts on DPPH free radical was no significant changed after spraying DPC. CONCLUSION: Plant growth regulator DPC could regulate the growth on the ground and underground effectively, and could enhance the content of flavonoids compounds of S. baicalensis.

[Effect of different developmental stage on plant growth and active compounds in Scutellaria baicalensis].

OBJECTIVE: To study the developmental phase on the growth and active compounds in Scutellaria baicalensis. METHOD: Seeds of wild plants were collected from Laiwu and sowed in Fangshan (Beijing) and Laiwu (Shandong). Samples of aerial and underground parts were collected in five growth periods of sprouts, seedlings, flowering, seed drop and withered periods respectively. The length of taproot, fresh weight of root, diameter of taproot and the length of stem were determined. The content of active compounds and total flavonoids were determined by HPLC and ultraviolet spectrophotometry respectively. The transcripted level of PAL1, PAL2, PAL3, C4H, 4CL, CHS, GUS and UBGAT were analyzed with RT-PCR. RESULT: The results showed that the aerial part of S. baicalensis grew quickly before flowering stage, and the underground part grew mostly between the periods of flowering and withered. In the whole growing developmental periods, the content of total flavonoids was not changed significantly, the content of baicalin was increased gradually and the content of baicalein was decreased gradually. Expression level of PAL and 4CL was the highest in withered period, CHS was increased between flowering and seed drop and decreased in withered period. CONCLUSION: Seedlings and withered periods may be the key phase affecting the growth and active compounds in S. baicalensis.

[Different pruning to yield and quality of Lonicerea Japonicae Flos].

OBJECTIVE: To find out the influence of differences pruning on Lonicerea Japonicae Flos on it's yield and quality, and provide reference for reasonable pruning. METHOD: Using different pruning treatments, the growth and yield and chlorogenic acid and galuteolin contents of Lonicerea Japonicae Flos were measured and compared. And the source and sink theory were used to interpret the rationality of the best way. RESULT: There existed significant differences in the growth and branching and yield at differences pruning methods, but not including chlorogenic acid and galuteolin contents of Lonicerea Japonicae Flos. CONCLUSION: The best pruning treatments is in shearing.

[Effect of acupuncture for dysphagia after stroke based on fiberoptic endoscopic swallowing function evaluation].

OBJECTIVE: To observe the effect of acupuncture combined with regular treatment and swallowing function training on pharyngeal motor, sensory function and penetration-aspiration function in patients with dysphagia after stroke. METHODS: A total of 60 patients with dysphagia after stroke were randomly divided into a control group and an observation group, 30 patients in each group. Both groups were treated with conventional treatment and swallowing function training; in addition, the observation group was treated with acupuncture at Lianquan (CV 23), Fengfu (GV 16), Yifeng (TE 17). All the treatments were given once a day, 5 days a week, for totally 4 weeks. In the two groups, the pharyngeal motor and sensory function, penetration-aspiration scores were evaluated by fiberoptic endoscopic evaluation of swallowing (FEES), and the Kubota water swallowing test scores were assessed before and after treatment, and the clinical effects were compared. RESULTS: After treatment, the pharyngeal motor and sensory function in the two groups were all higher than those before treatment (P<0.05), and those in the observation group were better than the control group (P<0.05). After treatment, the penetration-aspiration scores and Kubota water swallowing test scores in the two groups were all lower than those before treatment (P<0.05), and those in the observation group were lower than the control group (P<0.05). The total effective rate was 93.3% (28/30) in the observation group, which was better than 73.3% (22/30) in the control group (P<0.05). CONCLUSION: Acupuncture combined with regular treatment and swallowing training could improve the pharyngeal motor and sensory function, and penetration-aspiration scores in patients with dysphagia after stroke.

A Systematic Review and Meta-analysis on the Effectiveness of Transcranial Direct Current Stimulation on Swallowing Function of Poststroke Patients.

OBJECTIVE: The purpose of this study was to investigate the therapeutic effects of transcranial direct current stimulation on swallowing function in poststroke patients. DESIGN: We searched for potentially eligible randomized controlled trials from electronic databases, including the PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang, and Chinese Science and Technology Periodical (VIP) databases, from their inception to January 15, 2021. All statistical analyses were performed using RevMan 5.4, and the standardized mean difference with 95% confidence intervals was estimated for the swallowing function outcomes and to understand the mean effect size. RESULTS: Ten studies involving 343 participants were included in this meta-analysis. The overall analyses demonstrated a significant effect size for swallowing function. Subgroup analyses suggested that both acute and chronic stroke patients showed significant effects on swallowing function after transcranial direct current stimulation. Furthermore, compared with sham stimulation, transcranial direct current stimulation anodal to the affected, unaffected, and bilateral hemispheres can produce a significant effect size for swallowing function in stroke patients. CONCLUSIONS: This meta-analysis showed that transcranial direct current stimulation is likely to be effective for the recovery of dysphagia in poststroke patients, in the acute or chronic phase, and that the effect of anodal transcranial direct current stimulation to unaffected hemispheres is larger.

Protective Effects of Oroxylin A on Retinal Ganglion Cells in Experimental Model of Anterior Ischemic Optic Neuropathy.

Nonarteritic anterior ischemic optic neuropathy (NAION) is the most common cause of acute vision loss in older people, and there is no effective therapy. The effect of the systemic or local application of steroids for NAION patients remains controversial. Oroxylin A (OA) (5,7-dihydroxy-6-methoxyflavone) is a bioactive flavonoid extracted from Scutellariae baicalensis Georgi. with various beneficial effects, including anti-inflammatory and neuroprotective effects. A previous study showed that OA promotes retinal ganglion cell (RGC) survival after optic nerve (ON) crush injury. The purpose of this research was to further explore the potential actions of OA in ischemic injury in an experimental anterior ischemic optic neuropathy (rAION) rat model induced by photothrombosis. Our results show that OA efficiently attenuated ischemic injury in rats by reducing optic disc edema, the apoptotic death of retinal ganglion cells, and the infiltration of inflammatory cells. Moreover, OA significantly ameliorated the pathologic changes of demyelination, modulated microglial polarization, and preserved visual function after rAION induction. OA activated nuclear factor E2 related factor (Nrf2) signaling and its downstream antioxidant enzymes NAD(P)H:quinone oxidoreductase (NQO-1) and heme oxygenase 1 (HO-1) in the retina. We demonstrated that OA activates Nrf2 signaling, protecting retinal ganglion cells from ischemic injury, in the rAION model and could potentially be used as a therapeutic approach in ischemic optic neuropathy.

Association between achievement of hemodialysis quality-of-care indicators and quality-of-life scores.

BACKGROUND: Incremental achievement of quality indicator goals has been associated with progressive improvement in mortality and hospitalization risk in hemodialysis (HD) patients. STUDY DESIGN: Descriptive cross-sectional study. SETTING & PARTICIPANTS: All 33,879 HD patients treated at Fresenius Medical Care North America facilities for >90 days with scorable 36-Item Short Form Health Survey responses from January 1, 2006, to December 31, 2006. PREDICTOR: We hypothesized that achieving up to 5 HD goals before the survey (albumin >or= 4.0 g/dL, hemoglobin of 11-12 g/dL, equilibrated Kt/V >or= 1.2, phosphorus of 3.5-5.5 mg/L, and absence of HD catheter) results in better self-reported quality of life (QoL). OUTCOMES & MEASUREMENTS: Distributions of Physical and Mental Component Summary (PCS/MCS) scores within and across quality indicator categories determined during the prior 90 days from survey date (compared using analysis of covariance and linear regression models, with adjustment for case-mix and each of the quality indicators). RESULTS: Incremental achievement of up to 5 goals was associated with progressively higher PCS and MCS scores (both P for trend < 0.001). Compared with patients meeting all 5 goals (n = 4,208; reference group), case-mix-adjusted PCS score was lower by 1.8 point with only 4 goals met (n = 11,785), 3.4 points for 3 goals (n = 10,906), 4.9 points for 2 goals (n = 5,119), 5.9 points for 1 goal (n = 1,592), and 7.8 points in the 269 patients who failed to meet any goal (each P < 0.001 vs the reference group). The corresponding decreases in case-mix-adjusted MCS scores were 1.0 point for 4 goals met, 1.7 point for 3 goals, 2.3 points for 2 goals, 3.0 points for 1 goal, and 4.7 points with no goal met, with each P < 0.001 compared with the MCS score from patients who achieved all 5 goals. LIMITATIONS: Potential residual confounding from unmeasured covariates. CONCLUSION: Patients progressively meeting more quality goals report incrementally better QoL. Further studies are needed to determine whether prospective achievement of quality goals will result in improved QoL for HD patients.

Transient hyperopia after intensive treatment of hyperglycemia in newly diagnosed diabetes.

PURPOSE: Our aim was to observe the transient hyperopia during the intense glucose reduction in patients with newly diagnosed diabetes and severe hyperglycemia. STUDY DESIGN: Consecutive cases were observed. RESULTS: Totally 4 men and 1 woman with a mean age of 48 years were enrolled. In the 4 patients who received insulin, the hyperopia developed at 4.2 days after the initiation of treatment on average and reached a peak at 11.7 days; they recovered at 64.0 days. The other subject who received oral hypoglycemia agents revealed a peak change at 17 days and recovered at 70 days. A broader hyperopic change of 6.25 dpt was found in the patient with high myopia (-16 dpt). No significant difference was observed in the corneal curvature, axial length, lens thickness or depth of the anterior chamber during the course. The stable value of the accommodation amplitude and lens thickness may indicate that the cause of refraction change was due to the alteration in the reflection index of the lens. CONCLUSION: Intensive glucose reduction may cause transient hyperopia changes in newly diabetic patients and results in blurred vision.

Homonymous central quadrantanopia caused by an extrastriate (v2/v3) infarction: a case report.

A 66-year-old Taiwanese aboriginal male had complained of right-side blurred vision for 2 months, especially when reading. He had a 10-year history of hypertension and cardiovascular disease. His best-corrected visual acuity was 20/25 in each eye. Ophthalmoscopy revealed asymmetrical cupping, but a normal disc. Humphrey perimetry showed an upper homonymous paracentral quadrantanopic defect. Brain magnetic resonance imaging showed an infarction in the left lower calcarine area over the extrastriate (V2/V3) cortical area and a narrowing of the left middle and posterior cerebral arteries due to severe arteriosclerosis.

[Preliminary study on variation pattern of Cistanche deserticola].

OBJECTIVE: Through analyze of shape, structure and content of effective components of Cistanche deserticola, the variation of "you cong rong" is discussed. METHOD: The color surface, texture, the size of pith and the arrange of vascular bundle was observed with freshed, dried drugs and the transverse section, the effective component of echinacoside and acteoside was analyzed with hplc and the amylose was analyzed with ultraviolet spectrophotometry. RESULT: The variation of "you cong rong" is obvious different with the common drug in shape and structure, the content of echinacoside is 2.5 times, acteoside is 3.8 times, amylose yield is 1.6 times and the content is 2 times of the common drugs. CONCLUSION: "you cong rong" is an independent variation pattern of C. deserticola, it has no relationship with the ecological and geographical conditions.

[Comparative research on vitality and characteristics of seeds of Achyranthes bidentata in different producing area].

OBJECTIVE: To study the germination property of the seeds of Achyranthes bidentata from different producing area, providing foundation for choosing the better seed resources. METHOD: The germination rate, germination energy, germination index, vigor and the characteristics of the seeds of A. bidentata from different producing area were compared, and the relative electric conductivity was measured. The characteristics of the seed were recorded through the Anymicro DSS YT-5M microscope digital camera, and the differences were analyzed. RESULT: Seed from Henan Boai and Hebei Anguo showed the better germination rate, germination energy, germination index and vigor. CONCLUSION: The seeds from both Henan Boai and Hebei Anguo showed the better quality.

[Studies on callus culture of Akebia trifoliata].

OBJECTIVE: To investigate the inductive factors of effect on Akebia trifoliata and establish culture method for A. trifoliata callus. METHOD: To study the possible effective factors of culture condition by comparing with different explantation, nutrient medium, pH, temperature, illumination, growth substance of plant and its ratio. RESULT: The inductivity of leaves was the highest about 87.5%, followed with the stem section and leafstalk; The inductivity of nutrient medium such as MS, B5 callus was higher than the ones like H, SH and the White callus amended one; It was found that low-grade Phvalue benefits the growth of callus. The experiment result showed that different pH showed little difference in quality. The best condition of culture was 25 degrees C in temperature. CONCLUSION: The best culture condition for callus was the leaves as explantation. The A. trifoliata callus culture's best inductive condition was MS +2, 4-D 4.0 mg x L(-1) + NAA 1.0 mg x L(-1) + KT 1.0 mg x L(-1) (pH 5.8), cultural temperature was 25 degrees C, cultivation was dark.