Objective: To investigate the effect of puerarin on the regulation of AMPK-mTOR signaling pathway to inhibit autophagy and alleviate focal cerebral ischemia reperfusion injury. Methods: Forty male Sprague-Dawley rats were randomly divided into four groups: Sham group, model group, puerarin low-dose (50 mg/kg) group and puerarin high-dose (100 mg/kg) group. Pretreatment with puerarin for 7 d, then the middle cerebral artery occlusion (MCAO) model was established 0.5 h after the last administration according to Longa’s method. After 1.5 h of ischemia and 24 h of reperfusion, the neurological deficit scores were assessed, the infarct volume was calculated by TTC staining. The formation of autophagosome was observed by electron microscopy. The expression levels of LC3, p62, AMPK, p-AMPK, mTOR, p-mTOR, Ulk1, and pS757-Ulk1 were detected by Western blotting. Results: Compared with the Sham group, the neurological deficit scores and infarct volume in model group were significantly increased, the numbers of autophagosome increased, and the rate of LC3-II/LC3-I significantly increased, the expression level of p62 gradually decreased. The expression of p-AMPK was markedly up-regulated, while the expression of p-mTOR and pS757-Ulk1 was significantly down-regulated. Compared with the model group, the neurological deficit scores and infarct volume were significantly reduced, the number of autophagosome and the rate of LC3-II/LC3-I decreased, the expression of p62 was significantly up-regulated, the expression of p-AMPK was markedly down-regulated, the levels of p-mTOR and pS757-Ulk1 were significantly up-regulated. Conclusion: Puerarin alleviates cerebral ischemia-reperfusion injury may through suppressing autophagy via the AMPK-mTOR-Ulk1 signaling pathway.

[Summary] It has been believed that both environmental and genetic effects play roles in the pathogenesis of type 2 diabetes. Genetic factors may influence the effect of environmental factors on risk of developing type 2 diabetes. This review focused on the latest evidence of the interaction effect of genes and the environmental factors on type 2 diabetes.

Excessive autophagy is one of the crucial factors of cerebral ischemia-reperfusion injury (CIRI), which has been demonstrated to be one of the targets for acupuncture treatment of ischemic stroke. In the present paper, we make a review about the development of acupuncture intervention induced improvement of CIRI (such as reducing the infarction area, improving learning-memory ability and motor function) by regulating autophagy in animal studies. Outcomes showed that acupuncture intervention can function in 1) inhibiting CIRI-induced increase of the number of lysosomes and autophagic lysosomes, and relieving structural injury of mitochondria, and reducing the number of autophagosome in the central region of the ischemic cerebral cortex tissue; 2) down-regulating the expression of microtubule-associated protein Ⅱ light chain 3 (LC3Ⅱ) and the ratio of LC3-Ⅱ/LC3-Ⅰ in the ischemic cerebral region, and 3) regulating the expression of Beclin 1 (autophagy-related gene), promoting the expression of P62 (autophagy-related adaptor protein). In addition, acupuncture can also regulate phosphoinositide 3 kinase (PI3K)- protein kinase B (AKT)- mammalian target of rapamycin complex 1(mTOR) signaling at different time-points (down-regulation at the early stage and up-regulation at the later stage), and activating AMP-activated protein kinase (AMPK)-mTOR- UNC51-like kinase-1 signaling to relieve cerebral ischemic injury. These results reveal some mechanisms of acupuncture therapy underlying improvement of CIRI and provide experimental basis for clinical application of acupuncture therapy in the treatment of ischemic stroke.

To establish a new method for identifying genus of Lilium by DNA barcoding technology, ITS, ITS2, psbA-trnH, matK and rbcL sequences were analyzed in term of variation of inter- and intra-species, barcoding gap, neighbor-joining tree to distinguish genus of Lilium based on 978 sequences from experimental and GenBank database, and identification efficiency was evaluated by Nearest distance and BLAST1 methods. The results showed that DNA barcoding could identify different species in genus of Lilium. ITS sequence performed higher identification efficiency, and had significant difference between intra- and inter-species. And NJ tree could also divide species into different clades. Results indicate that DNA barcoding can identify genus of Lilium accurately. ITS sequence can be the optimal barcode to identify species of Lilium.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Lilium ; classification

The excess fatty tissues on the head, neck, thorax, and abdomen of morbid obese patients can impede the patency of the upper airway and impair lung functions. Meanwhile, these patients often have comorbidities such as obstructive sleep apnea, hypoventilation syndrome, chronic obstructive pulmonary disease, and asthma, which may result in difficult airway, intraoperative hypoventilation, and postoperative respiratory depression. Therefore, perioperative airway management for morbid obese patients may pose a big challenge to anesthesiologists. Anesthesiologists should know well about the pathophysiological features of respiratory system and grasp rational management principles, so as to improve the safety and effectiveness of perioperative airway management and optimize the clinical prognosis.
Airway Management ; Anesthesia ; methods ; Humans ; Obesity ; surgery ; Perioperative Care

Objective To evaluate the association of islet autoantibodies [ glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase antibody(IA-2A)and insulin autoantibodies(IAA)1 with HLA- DQ genotypes in the first-degree relatives of autoimmune type 1 diabetes mellitus.Methods This was a cross- sectional and case-control study.Three hundred and fifty-one first-degree relatives with normal glucose tolerance of patients with type 1 diabetes mellitus and 376 healthy controls were recruited and measured for GADA,IA-2A and IAA by radioligand assay,and 156 first-degree relatives of patients with autoimmune type 1 diabetes mellitus and 278 controls were typed for genetic polymorphisms of HLA-DQ with PCR sequencing-based typing method.Results (1)DQA1*03,DQBI*0303,*0401 alleles and DQA1 * 03-DQBI * 0303,DQA1 * 05-DQBI * 0201,DQA1 * 03-DQBI * 0401 haplotypes were significantly increased in the first-degree relatives of autoimmune type 1 diabetes mellitus(P

Helicobacter pylori was separated from gastric mucosa of chronic gastritis patients for the first time in 1983, and it is the only species known to be able to survive in the human stomach. The Urase, Vacuolating cytotoxin, cytotoxin, adhesin, catalase, Outer Inflammatory Protein, duodenal ulcer promoter factor and lipopolysaccharide antigen of HP were reviewed in this paper, so that scholars can understand the recent research situation of HP antigen. The above study lays the foundation for future screening of high efficient antigen and increasing the expression of antigen, and exploring new methods of epitope binding in HP epitope vaccine.

OBJECTIVETo investigate the expression changes of metabotropic glutamate receptor 5 (mGluR5) in neuropathic pain.

METHODSEighty-four adult male Sprague Dawley rats weighing 180-220 g were randomly divided into 7 groups (n = 12) : control group; S3, S7, and S14 groups: rats received the sham operation, the mechanical pain threshold was measured, and then the rats were decapitated and the ipsilateral lumbar spinal cord dorsal horn and dorsal root ganglion (DRG) samples were obtained on the 3rd, 7th, 14th postoperative day, respectively; C3, C7, and C14 groups: the chronic sciatic nerve constriction (CCI) model was established, the mechanical pain threshold was measured and the samples were obtained on the 3rd, 7th, 14th postoperative day, respectively. The expression level of mGluR5 mRNA and protein in the spinal cord and DRG were measured using the reverse transcriptase polymerase chain reaction and Western blot.

RESULTSIn the CCI group, the mechanical pain threshold in each observation day was significantly lower than in the sham operation group (P < 0.05). In the spinal cord, the expressions of mGluR5 mRNA and protein were significantly elevated in the C3 group than in the S3 and the control group (P < 0.05). On the 7th and the 14th postoperative day, no significant difference was found in the expression of mGluR5 mRNA and protein between CCI groups and the sham operation groups or the control group. No change was detected in DRG mRNA or protein.

CONCLUSIONmGluR5 is differentially expressed in spinal cord in response to neuropathic pain, which suggests that mGluR5 may be involved in the mechanism of neuropathic pain.

Animals ; Disease Models, Animal ; Ganglia, Spinal ; metabolism ; Male ; Neuralgia ; metabolism ; Pain Threshold ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, Metabotropic Glutamate 5 ; Receptors, Metabotropic Glutamate ; genetics ; metabolism ; Sciatic Neuropathy ; metabolism ; Spinal Cord ; metabolism

AIMTo identify "Shegan" [Belamcanda chinensis (L.) DC.] and relative medicinal plants of Iris including Iris tectorum Maxim., I. dichotoma Pall., I. germanica L. and I. japonica Thunb. by ribulose 1,5-bisphosphate carboxylase Large Gene (rbcL) sequence analysis.

METHODSGeneral DNA was isolated from the fresh leaves of Belamcanda chinensis and 4 Iris spp. by CTAB. A pair of primers was designed to amplify the rbcL gene and PCR Preps DNA kit was used to purify the PCR products. The rbcL sequences were determined by ABI (Applied Biosystems Inco.) Prism 310 Genetic Analyzer.

RESULTSA fragment of about 750 bp of rbcL gene from Belamcanda chinensis and 4 Iris spp. were amplified and sequenced. The rbcL sequences of Iris tectorum, I. dichotoma Pall. and I. japonica were reported for the first time. The rbcL sequences of 5 species of Iridaceae were aligned and analyzed using Clustal (Version 8.0) and MEGA (Version 2.0.) programs. The nucleotide number of difference is from 1.000 to 20.000. The tranversions is from 0.000 to 9.000 and the transitions is from 0.000 to 14.000. Phylogenetic tree based on rbcL partial sequence data indicated that the eleven samples of 5 species clustered separately.

CONCLUSIONThe sequence variation of rbcL can be used to identify Belamcanda chinensis and 4 species of relative medicinal plants of Iris. The molecular phylogenetic tree accords with the classical taxonomy.

Base Sequence ; Chloroplasts ; genetics ; DNA, Plant ; analysis ; Genes, Plant ; Iridaceae ; classification ; genetics ; Iris Plant ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Ribulose-Bisphosphate Carboxylase ; classification ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo probe the mechanism of acupuncture in treatment of dysmenorrhea.

METHODSAdult mice with no pregnancy were randomly divided into a normal group, a model group, an acupuncture group and a medication group. The model group, the acupuncture group and the medication group were modeled by Diethylstilbestrol and Ocytocin. For the acupuncture group, at the 7th day of modeling, acupuncture was given at "Sanyinjiao" (SP 6), "Diji" (SP 8), once a day, for 5 days; and at the 7th day of modeling, Yimucao Gao 0.6 mg/g was given intragastrically to the medication group for 5 days. The stretching latent period and the number of stretching within 30 min were observed, and mRNA levels of ocytocin receptor (OctR) and vasopressin receptor (VasR) in the uterus tissue were detected with RT-PCR method.

RESULTSCompared with the model group, the stretching latent period extended (P < 0.05) and the number of stretching within 30 min significantly decreased (P < 0.05); and there were significant differences in the mRNA levels of ocytocin receptor and vasopressin receptor in the uterus tissue in the model group as compared with those in other 3 groups (P < 0.05, P < 0.01).

CONCLUSIONAcupuncture can improve the dysmenorrheal symptom to a certain extent, and the mechanism is possibly related to regulative effects of acupuncture on hormones-mediating receptors in mice.

Acupuncture Therapy ; Animals ; Dysmenorrhea ; metabolism ; therapy ; Female ; Mice ; RNA, Messenger ; analysis ; Receptors, Oxytocin ; genetics ; Receptors, Vasopressin ; genetics