Specific DNA barcodes screening, germplasm resource identification, and genetic diversity analysis of Platycodon grandiflorum / 药学学报

Platycodonis Radix is the dry root of Platycodon grandiflorum of Campanulaceae, which has a variety of pharmacological effects and is a commonly used bulk Chinese medicine. In this study, the chloroplast genome sequences of six P. grandiflorum from different producing areas has been sequenced with Illumina HiSeq X Ten platform. The specific DNA barcodes were screened, and the germplasm resources and genetic diversity were analyzed according to the specific barcodes. The total length of the chloroplast genome of 6 P. grandiflorum samples was 172 260-172 275 bp, and all chloroplast genomes showed a typical circular tetrad structure and encoded 141 genes. The comparative genomics analysis and results of amplification efficiency demonstrated that trnG-UCC and ndhG_ndhF were the potential specific DNA barcodes for identification the germplasm resources of P. grandiflorum. A total of 305 P. grandiflorum samples were collected from 15 production areas in 9 provinces, for which the fragments of trnG-UCC and ndhG_ndhF were amplificated and the sequences were analyzed. The results showed that trnG-UCC and ndhG_ndhF have 5 and 11 mutation sites, respectively, and 5 and 7 haplotypes were identified, respectively. The combined analysis of the two sequences formed 13 haplotypes (named Hap1-Hap13), and Hap4 is the main genotype, followed by Hap1. The unique haplotypes possessed by the three producing areas can be used as DNA molecular tags in this area to distinguish from the germplasm resources of P. grandiflorum from other areas. The haplotype diversity, nucleotide diversity and genetic distance were 0.94, 4.79×10-3 and 0.000 0-0.020 3, respectively, suggesting that the genetic diversity was abundant and intraspecific kinship was relatively close. This study laid a foundation for the identification of P. grandiflorum, the protection and utilization of germplasm resources, and molecular breeding.

Screening of specific DNA barcode, identification of germplasm resources, and analysis of genetic diversity of Atractylodes chinenesis / 药学学报

italic>Atractylodes chinensis has important medicinal and economic values. In this study, the chloroplast genome sequences of four A. chinensis samples from different producing areas were sequenced using the Illumina platform. The specific DNA barcodes were screened and the germplasm resources of A. chinensis samples from different producing areas and the genetic diversity of the population were analyzed basing on the specific barcodes. The whole chloroplast genomes of the four A. chinensis samples had a typical cyclic tetrad structure, with 112 genes annotated. The comparative genomics results indicated that ccsA and trnC-GCA_petN were potential specific DNA barcodes for intraspecific identification of A. chinensis. Polymerase chain reaction (PCR) analysis of ccsA and trnC-GCA_petN was performed on 256 samples from 14 areas in 9 provinces, and the amplification efficiency was 100%. Sequence analysis showed that ccsA and trnC-GCA_petN had 11 and 22 variant positions, which could identify 16 and 22 haplotypes, respectively. The combined sequence analysis identified 39 haplotypes, named Hap1-Hap39, of which the most abundant and widely distributed genotype was Hap9. Haplotype diversity (Hd) = 0.896 and nucleotide diversity (Pi) = 0.002 22 indicated high genetic diversity at the species level in A. chinensis. The genetic distances of the haplotypes were 0.000 00-0.004 88, indicating that there were small genetic differences among the haplotypes. The results of phylogenetic tree analysis showed that 39 haplotypes had very close genetic relationship, and formed two obvious branches with other groups of the same genus except Atractylodes macrocephala. This study plays an important role in the identification of the origin of A. chinensis and the protection and breeding of germplasm resources.

Effect of fucoxanthin on insulin resistance in obese mice induced by high fat diet / 中国中药杂志

The aim of this paper was to study the effect and mechanism of fucoxanthin on insulin resistance of obese mice induced by high-fat diet. Fifty C57 BL/6 J male mice were randomly divided into control group and high-fat diet group. The insulin resistance model was induced with high-fat diet for 12 weeks, and model mice were randomly divided into model group, fucoxanthin-0.2% group, fucoxanthin-0.4% group and metformin group. After dietary treatment for 6 weeks, the body weight and epididymal fat weight in each group were measured. Fasting blood glucose(FBG), fasting insulin(FINS), total cholesterol(TC), triglyceride(TG), low-density lipoprotein(LDL-C) and high-density lipoprotein(HDL-C) were measured, and insulin resistance index(HOMA-IR) was calcula-ted. The pathological morphology in liver was observed by hematoxylin eosin staining, and the expressions of some key proteins in insulin receptor substrate 1(IRS-1)/posphoinositide 3-kinase(PI3 K)/serine-threonine kinase(Akt) and peroxisome proliferators-activated receptor-γ(PPARγ)/sterol regulatory element binding protein-1(SREBP-1)/fatty acid synthetase(FAS) pathways in liver were detected by Western blot. According to the findings, compared with the model group, levels of body weight, epididymal fat weight, FBG, FINS, TC, TG, LDL-C and HOMA-IR, as well as protein expressions of PPARγ, SREBP-1 and FAS in liver were significantly reduced(P<0.05 or P<0.01), while level of HDL-C and protein expressions of p-IRS-1, IRS-1, PI3 K and p-Akt in liver were signi-ficantly increased after treatment with fucoxanthin(P<0.05 or P<0.01). And the pathological changes of liver tissue in fucoxanthin-treated mice were also improved obviously. The results showed that fucoxanthin could improve obesity, hyperglycemia and hyperlipidemia, and alleviate insulin resistance in obese mice, and its mechanism is possibly related to the regulation of IRS-1/PI3 K/Akt and PPARγ/SREBP-1/FAS pathways.

Lung transplantation as therapeutic option in acute respiratory distress syndrome for coronavirus disease 2019-related pulmonary fibrosis / 中华医学杂志(英文版)

BACKGROUND@#Critical patients with the coronavirus disease 2019 (COVID-19), even those whose nucleic acid test results had turned negative and those receiving maximal medical support, have been noted to progress to irreversible fatal respiratory failure. Lung transplantation (LT) as the sole therapy for end-stage pulmonary fibrosis related to acute respiratory distress syndrome has been considered as the ultimate rescue therapy for these patients.@*METHODS@#From February 10 to March 10, 2020, three male patients were urgently assessed and listed for transplantation. After conducting a full ethical review and after obtaining assent from the family of the patients, we performed three LT procedures for COVID-19 patients with illness durations of more than one month and extremely high sequential organ failure assessment scores.@*RESULTS@#Two of the three recipients survived post-LT and started participating in a rehabilitation program. Pearls of the LT team collaboration and perioperative logistics were summarized and continually improved. The pathological results of the explanted lungs were concordant with the critical clinical manifestation, and provided insight towards better understanding of the disease. Government health affair systems, virology detection tools, and modern communication technology all play key roles towards the survival of the patients and their rehabilitation.@*CONCLUSIONS@#LT can be performed in end-stage patients with respiratory failure due to COVID-19-related pulmonary fibrosis. If confirmed positive-turned-negative virology status without organ dysfunction that could contraindicate LT, LT provided the final option for these patients to avoid certain death, with proper protection of transplant surgeons and medical staffs. By ensuring instant seamless care for both patients and medical teams, the goal of reducing the mortality rate and salvaging the lives of patients with COVID-19 can be attained.

Effects of pien-tze-huang on expression of NMDAR1 and GluR2 in cortex of MCAO rats / 中国药理学通报

Aim: To investigate the effects of Pien-Tze-Huang (PZH) on mRNA and protein expression of NMDAR1 and GluR2 in cortex of MCAO rats. Methods: Forty healthy adult male Sprague-Dawley rats were randomly divided into three groups: sham, MCAO, MCAO + PZH groups. The rats were subjected to focal cerebral ischemia/reperfusion with suture-occluded method. Neurological deficit testing was performed with Zea Longa scale. The volume of cerebral infarction was evaluated by TTC staining. The mRNA and protein expression of NMDAR1 and GluR2 in cortex of side cerebral ischemic tissues were determined using qPCR and Western blot analysis. Results: Compared with MCAO group, PZH significantly improved the neurological deficit, decreased the volume of cerebral infarction, and up-regulated the mRNA and protein expression of NMDAR1 and GluR2. Conclusions: PZH attenuates the down-regulation of mRNA and protein expression of NMDAR1 and GluR2 after focal cerebral ischemic injury in rats, which may be associated with the cerebral protective effect of PZH.

Association of Cosmc gene mutation with susceptibility to Henoch-Schönlein purpura in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the presence of Cosmc gene mutation in children with Henoch-Schönlein purpura (HSP) and the association between Cosmc gene mutation and the susceptibility to HSP.</p><p><b>MESULTS</b>Eighty-four children who were diagnosed with HSP between March 2014 and December 2015 were selected as the HSP group. Fifty-eight healthy volunteers matched for age and sex were enrolled as the control group. Fasting venous blood (5 mL) from the two groups was collected in EDTA anticoagulated tubes, followed by the isolation of peripheral blood mononuclear cells (PBMCs) through density gradient centrifugation. Genomic DNA was extracted from PBMCs according to the manufacturer's protocol, and the whole exon region of Cosmc gene was amplified by touch-down polymerase chain reaction (touch-down PCR). The PCR products were identified by 1% agarose gel and sequenced in order to further examine the association between Cosmc gene mutation and the susceptibility to HSP.</p><p><b>RESULTS</b>Sequencing results showed two mutations (c.393T>A and c.72A>G) of Cosmc gene in children with HSP. There were no significant differences in the genotype and allele frequencies at the two loci between the HSP and control groups, and this distribution was not associated with sex.</p><p><b>CONCLUSIONS</b>The mutations c.393T>A and c.72A>G in the exon region of Cosmc gene in children with HSP are not associated with the onset of HSP.</p>

The role of oxidative inflammatory cascade on pancreatic fibrosis progression in mice induced by DBTC plus ethanol / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the role and mechanism of oxidative inflammatory cascade in pancreatic fibrosis progression of chronic pancreatitis (CP) in mice induced by dibutyltin dichloride (DBTC) plus ethanol.</p><p><b>METHODS</b>Thirty-six KM mice were randomly divided into 2 groups (n = 18): control group and model group (DBTC combined with ethanol). The mice in model group were intravenously injected with DBTC (8 mg/kg) in tail vein and drink 10% ethanol. After modeling 2 weeks, 4 weeks and 8 weeks, the mice were anesthetized and sacrificed, the pathological changes and the degree of fibrosis in the pancreas were observed by HE and Masson staining, the F4/80 expression level were detected by immunohistochemistry, the content of superoxide dismutase (SOD), malondialdehyde(MDA) and myeloperoxidase (MPO) were measured in the pancreatic homogenates.</p><p><b>RESULTS</b>The fibroblasts and macrophages (f4/80 positive staining) could be seen obviously in pancreas of model group at 2 weeks. At 4 weeks and 8 weeks, macrophages infiltration increased and pancreatic tissue was substituted by the proliferation of fibrosis significantly. At every time-point, in pancreatic homogenates SOD was decreased, MDA and MPO markedly increased. There was significant differences between two groups (P < 0.05).</p><p><b>CONCLUSION</b>DBTC injection joint ethanol drinking can successfully establish the model of chronic pancreatitis and pancreatic fibrosis in mice. Oxidative inflammatory cascade plays an important role in the progression of pancreatic fibrosis.</p>

A new herbs traceability method based on DNA barcoding-origin-morphology analysis--an example from an adulterant of 'Heiguogouqi' / 中国中药杂志

The fruit of Lycium ruthenicum is a common folk medicine in China. Now it is popular for its antioxidative effect and other medical functions. The adulterants of the herb confuse consumers. In order to identify a new adulterant of L. ruthenicum, a research was performed based on NCBI Nucleotide Database ITS Sequence, combined analysis of the origin and morphology of the adulterant to traceable varieties. Total genomic DNA was isolated from the materials, and nuclear DNA ITS sequences were amplified and sequenced; DNA fragments were collated and matched by using ContingExpress. Similarity identification of BLAST analysis was performed. Besides, the distribution of plant origin and morphology were considered to further identification and verification. Families and genera were identified by molecular identification method. The adulterant was identified as plant belonging to Berberis. Origin analysis narrowed the range of sample identification. Seven different kinds of plants in Berberis were potential sources of the sample. Adulterants variety was traced by morphological analysis. The united molecular identification-origin-morphology research proves to be a preceding way to medical herbs traceability with time-saving and economic advantages and the results showed the new adulterant of L. ruthenicum was B. kaschgarica. The main differences between B. kaschgarica and L. ruthenicum are as follows: in terms of the traits, the surface of B. kaschgarica is smooth and crispy, and that of L. ruthenicum is shrinkage, solid and hard. In microscopic characteristics, epicarp cells of B. aschgarica thickening like a string of beads, stone cells as the rectangle, and the stone cell walls of L. ruthenicum is wavy, obvious grain layer. In molecular sequences, the length of ITS sequence of B. kaschgarica is 606 bp, L. ruthenicum is 654 bp, the similarity of the two sequences is 53.32%.

Acquisition and storage methods for image data collected in Chinese medicine resources survey / 中国中药杂志

The acquisition and storage of the image data are important in the Chinese medicine resources survey, and it is important data and evidence for the process and the results. The image data of the Chinese medicinal materials' habitat, original plant or animal, processing in habitat, commodity form, the relative contents and workshop scenarios in the investigation are important for the compiling of the Color Atlas of National Chinese Medicine Resources, mapping the digital scattergram of the Chinese medicine resources, establishing the digital Chinese medicine plant herbarium and acquiring the documentary of the Chinese medicine resource survey. The content, procedures and methods of the video data collecting have been related and analyzed in this article to provide reference for the Chinese medicine resources survey.

Gelsolin level for prognostic evaluation in patients with critically illness / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the changes of plasma gelsolin level in patients with critical illness and its application in prognostic evaluation.</p><p><b>METHODS</b>Ninety six critically ill patients admitted in ICU of Sichuan Provincial People's Hospital from February 2012 to December 2013 were enrolled in the prospective cohort study. Plasma gelsolin levels were detected with enzyme linked immunosorbent assay (ELISA) at admission (d1), d2, d4 and d8 after admission, and also detected in blood samples of 186 healthy subjects as controls. Logistic regression model was used to analyze the relationship between the level of plasma gelsolin and prognosis of patients.</p><p><b>RESULTS</b>The average levels of plasma gelsolin were significantly lower in critically ill patients than those in control subjects (F=1986.37, P<0.01). There was significant difference in overall level of gelsolin between survival patients and fatal patients (F=16.691, P<0.01). APACHE Ⅱ score was associated with survival outcomes (r=0.489, P=0.009); the APACHE Ⅱ score was significantly higher in fatal patients than that in survival patients (29.5±7.7 vs 22.1±5.7, t=5.375, P<0.01). There was a negative correlation between plasma gelsolin levels and fatal outcomes (r=-0.512, P<0.01). Logistic regression analysis showed that the overall plasma gelsolin levels and the last measured level was a prognostic factor for critically ill patients (P<0.05).</p><p><b>CONCLUSION</b>Plasma gelsolin levels are correlated with the severity of critically ill patients, and plasma gelsolin can be used as indicator of prognosis.</p>

Psychological distress, family functioning, and social support in family caregivers for patients with dementia in the mainland of China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Dementia is a chronic progressive disease seriously affecting the patient's daily life and working skills and may cause the patient disability and dependence. Thus, caring for dementia patients inevitably falls on families in the mainland of China. Unfortunately, there are rarely enough reports available about mental health and social support in family caregivers for domestic dementia patients. This study aimed to investigate the changes in psychological status and social support in domestic dementia family caregivers and hope the government and relevant departments pay more attention to the family caregiver's psychological change, which may be better for dementia patients.</p><p><b>METHODS</b>Fifty-eight immediate family caregivers (family group) for dementia patients, including 21 Alzheimer's disease (AD) and 37 vascular dementia inpatients, were recruited for interview. Fifty-eight age- and sex-matched normal volunteers (control group) were also interviewed in the same period. The psychiatric distress was assessed by the Symptom Checklist 90 (SCL-90), family functioning was assessed by the Family Assessment Device (FAD), and the social support was assessed by the Multidimensional Scale of Perceived Social Support (MSPSS). Group differences were analyzed using unpaired t test for comparison of SCL-90, FAD, and MSPSS mean scores. Pearson's correlation coefficient was used to find the association between the various dimensions of FAD and the social support from dementia family caregivers.</p><p><b>RESULTS</b>Except phobic anxiety dimension, the other dimensions of SCL-90 mean scores were significantly higher in the family group than those in the control group (P < 0.05). In the family group, except affective involvement dimension, the other dimensions of FAD mean scores were within the scope of unhealthy family functioning. Significant differences in problem solving, communication, roles, affective responsiveness, behavioral control, general functioning, and the total MSPSS mean scores were seen between the family group and the control group (P < 0.01). Except affective involvement dimension, the other dimensions of the family functioning correlated with the perceived social support in the family group (P < 0.05 or 0.01).</p><p><b>CONCLUSIONS</b>Dementia patients have a negative effect on families' mental health in the mainland of China. Dementia family caregivers experience defected family functioning and low levels of social support.</p>

Sequence analysis and identification of a chloroplast matK gene in Rhei Rhizoma from different botanical origins / 药学学报

Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.

Efficacy of sequential ablation of sinus atrial node fat pad and atrial ventricular node fat pad on inducibility of atrial fibrillation evoked by vagus trunk stimulation / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the efficacy of sequential ablation of epicardial fat pad on inducibility of atrial fibrillation (AF) evoked by stimulating vagus trunk.</p><p><b>METHODS</b>Eighteen adult mongrel dogs were randomly divided into 2 groups (n = 9 each): Group A underwent pre-ablation of sinus-atrial node fad pad (SANFP) and subsequent ablation of atria-ventricular node fad pad (AVNFP). Group B underwent pre-ablation of AVNFP and subsequent ablation of SANFP. AF was induced by high-frequency electrical stimulation of bilateral vagus trunks. The AF inducibility and effective refractory period (ERP) changes during vagus trunk stimulation were examined before and after ablation in atria and pulmonary veins.</p><p><b>RESULTS</b>(1) AF could be induced by vagus trunk stimulation and the incidence was higher during right vagus trunk (RVG) stimulation than left vagus trunk (LVG) stimulation [(60.0 ± 0.0)% vs (18.4 ± 22.1)%]. (2) SANFP ablation significantly attenuated AF inducibility with LVG stimulation and RVG stimulation at 2 V (decreased 67.0% and 72.0%, respectively). Subsequent AVNFP ablation after SANFP ablation further reduced AF inducibility with LVG and RVG stimulation at 2 V (decreased 100.0% and 95.5%, respectively). (3) AVNFP ablation (decreased 95.7% and 96.3%, respectively) and subsequent SANFP ablation after AVNFP ablation (decreased 98.0% and 100.0%, respectively) significantly attenuated AF inducibility with LVG stimulation and RVG stimulation at 2V. (4) Vagal stimulation induced ERP shortening was significantly attenuated by isolated SANFP ablation or AVNFP. Subsequent AVNFP ablation after SANFP induced significant ERP shortening in right atrial site compared with isolated SANFP ablation. However, changes of ERP shortening were similar between AVNFP ablation and subsequent SANFP ablation after AVNFP ablation.</p><p><b>CONCLUSIONS</b>Epicardial fat pad ablation reduced the AF inducibility and prolonged ERP of atria and pulmonary veins during vagus trunk stimulation. AVNFP, as the "integration centers" modulating the vagal innervation to the atria, may be the more effective target of ablation for treating AF.</p>

Effects of life style intervention on obesity-related vascular dysfunction in children / 中华流行病学杂志

Objective To evaluate the reversibility of obesity-related arterial dysfunction and thickening carotid intima-media through dietary and/or exercise intervention programs.Methods Seventyeight 9 to 12 year-old overweight children,were randomly assigned to group A:only dietary modification,group B:diet plus a supervised structured exercise program for 6 weeks(SE)and subsequently for 1 year (CE).The prospectively defined primary end points were assessed through ultrasound-derived arterial endothelial function(endothelium-dependent dilation)of the brachial artery and intima-media thickness of common carotid artery.Results At 6 weeks,both interventions were associated with improved arterial endothelial function[group A:(6.36±0.81)% vs.(5.32±0.84)%,P＜0.01;group B:(7.46±0.91)% vs.(5.20±0.87)%,P＜0.01 ].A conbination of diet and exercise were associated with a significantly greater improvement in endothelial function than on diet alone(P＜0.01).At 1 year,there appeared significantly Less thickening of the carotid wall in group CE[(0.57±0.05)mm vs.(0.59±0.07)mm,P＜0.001].Compared with children in group SE[(6.38±1.04)% vs.(5.22±0.96)%,P＜0.05],vascular function appeared significantly better in group CE[(8.29±0.78)% vs.(5.38±0.65)%,P＜0.001].Conclusion Obesity-related vascular dysfunction in young children was partially reversible with diet alone but more obvious when combined with exercise training for 6 weeks.Sustained improvements at 1 year was seen in those who persisted in diet program plus regular exercise.