Endoscopy ; methods ; Epidermal Cyst ; surgery ; Female ; Humans ; Middle Aged ; Skull Base ; pathology

Adult ; Chronic Disease ; Female ; Humans ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Nasal Polyps ; etiology ; Postoperative Complications ; Sinusitis ; etiology

AIM:To study the application of ultrasonic extraction in process of extracting diosgenin from Dioscorea zingiberensis C.H.Wright,and to evaluate its merits.METHODS:A new process of ethanol extracting and hydrochloric acid hydrolysis was provided to extraction diosgenin,and five different extraction methods were studied and compared,and the reaction mechanism was discussed.RESULTS:The ultrasonic extraction method was more efficient.Compared with the traditional water-hydrolylis,extraction rate was 15 percent higher,dosage of hydrochloric acid was registered 90 percent decrease,and the production was purer.CONCLUSION:The grinding pretreatment of Dioscorea zingiberensis is help to enhance the effects of ultrasonic extraction,the method has some advantages in efficiency,energy-saving and low pollution.

Background Laser assisted subepithelial keratomileusis (LASEK) is one of surgical procedures for refractive correction.Dilute alcohol that is used for the removal of epithelium during LASEK induces the apoptosis of corneal epithelial cells.Researches showed that thymosin β4 (Tβ4) can arrest apoptosis, but whether Tβ4 plays inhibitory effect on ethanol-induced damage of corneal epithelial cells is still unelucidated.Objective The aim of this study was to investigate the protective effects of Tβ4 on ethanol-induced rabbit corneal epithelial cell damage in vitro.Methods The corneal tissue of deendothelium was obtained from 10 New Zealand white rabbits.Corneal epithelial cells were cultured in vitro by using explant culture method.Cultured cells were identified by detecting the expression of keratin 12 and connexin 43 with reverse transcription PCR (RT-PCR).The cells of second generation were collected and divided into 4 groups.The cells were regularly cultured in the normal control group, and Tβ4 was added in the culture medium at the final concentration of 1 μg/ml in the Tβ4 treated group.Ethanol-induced rabbit corneal epithelial cell damage models were established by adding PBS containing 20％ alcohol in medium for 20 seconds in the model group,and Tβ4 was added in the medium of model cells at the final concentration of 1 μg/ml in the model+Tβ4 group.The survival rate of the cells was detected by MTT assay, and the apoptosis rate of the cells was examined by TUNEL method.The relative expression levels of cyclin D1 mRNA and cyclin-depensent protein kanase 4 (CDK4) mRNA in the cells were detected by real-time flurescenee quantitative PCR.The content of bcl-2 protein in the cells was detected by ELISA assay.Spectrophotometry was employed to measure the activity of caspase-3.The study complied with the Regulation for the Adminstration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The mean cell survival rate was (52.1 ± 14.07) ％ in the model group,which was significantly reduced in comparison with 100％ of the normal control group and (77.7± 19.60) ％ of the model+Tβ4 group (P=0.001 ,P=0.035).TUNEL staining revealed more positive cells in the model group and the model+Tβ4 group,and the percentage of TUNEL positive cells was (30.0±6.7)％ in the model+Tβ4 group, showing an evident decline in comparison with (42.4±4.0)％ of the model group (P=0.01).The relative expression levels of cyclin D1 mRNA and CDK4 mRNA were 0.93±0.17 and 0.88±0.09 in the model+Tβ4 group, which were significantly higher than 0.68±0.05 and 0.54±0.07 in the model group (P=0.027,0.002).ELISA assay exhibited that bcl-2 content in the cells was considerably lower,and caspase-3 activity was significantly higher in the model group than those in the model+Tβ4 group (P =0.030,0.021).Conclusions Tβ4 plays a protective effect on rabbit corneal epithelial cells from apoptosis by lowing the caspase 3 activity and increasing bcl-2 content in ethanoldamaged rabbit corneal epithelial cells.In addition, Tβ4 promotes the regrowth of corneal epithelial cells by upregulating the expression of cyclin D1 and CDK4.

Opioid receptors have drawn highly attention to scientists since they were founded in the 1990s.As a member of G protein-coupled receptor family,opioid peptides are the endogenous ligands.It is well known to us that the basic structure consists of extracellular amino residues,seven transmembrane region and the hydroxyl residues of the cell.In recent years,the progress of science laid the foundation for further studies on the function of delta receptor.Aberrant expression of delta opioid receptor in a variety of tumors,and it plays an important role of the occurrence and development of tumors.This review describes the recent research advances on delta opioid receptor and its association with hepatocellular carcinoma.

Objective To obtain the soluble single-chain fragment V (ScFv)monoclonal antibodies (McAbs) against the SSA antigen epitopes.Methods Three octapeptides (60 000 SSA antigen residues 482-493 termed as P1 epitope, residues 310-323 termed as P2 epitope and residues 230-241 termed as P3 epitope) were synthesized on the lysine frame.The McAbs were panned by coating the corresponding as targets.The specificity, affinity and gene squences of the positive clones were assessed.Soluble single-chain fragment V antibodies special for SSA antigen epitopes were expressed and then identificated.Results After 5 rounds of panning, reactive scFv clones contained full-length scFv antibodies coding regions were obtained,with sufficient affinity and specificity for respective antigen peptides.The absorbance values at 410 nm of the fusion protein of anti-P1-P3 activity with the corresponding peptides were 1.43 ± 0.23, 0.82 ±0.31 and 0.80 ± 0.25, and there was also statistically significant difference in the cross reactions ( P ＜ 0.01 ).Three clones were successfully expressed and then purified by His-bind resin.The activity in vivo of soluble ScFv antibodies was identified to be positive by the indirect immune-fluorescence assay on Hep-2 cells.Conclusion Souble ScFv McAbs against corresponding SSA antigen peptides with high affinity,specificity and activity in vivo were obtained, which are to be competent enough for epitopes expression on the target organs.

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

Objective To explore a method of establishing a canine model of intracerebral hemorrhage (ICH) by puncturing the main branch of middle cerebral artery (MCA) under ultrasonic guidance and proceed a pilot study of contrast-enhanced ultrasound (CEUS) of ICH.Methods Twelve adult healthy mongrel dogs were enrolled in the experiment.In the condition of bone window,the main branch of MCA was punctured to bleed under the ultrasonic guidance.CEUS of brain was performed every 30 minutes until hematoma formed and another CEUS was performed at 24 hours.Then brains were collected for pathologic examination after dogs were euthanized.Results Eleven of twelve dogs developed ICH successfully that located in the Out edge of basal ganglia,which was confirmed by CEUS and autopsy.One dog died within 6 hours due to intraventricular hemorrhage which was confirmed by autopsy.The hematoma diameters were measured as (22.4 ± 7.1)mm by CEUS before euthanasia and as (21.6 ± 6.9)mm on autopsy.There was no significant difference between the two methods of measurement (t =1.521,P =0.1565).The characteristics of active bleeding and hematoma on ICH were performed in the CEUS imaging.Conclusions A model of dog ICH by puncturing the main branch of MCA under the guidance of ultrasound was established successfully,and it proved to be simple,effective and repeatable.The imaging characteristics of this model are in good accordance with those of ICH in patients.

In traditional oral practice, the presystemic interactions with gut microbiota is an important mechanism underlying the holistic health benefits of Chinese herbal medicines (CHMs), making the study of CHMs distinct from the research of Western medicines of which the systemic exposure (level in blood) is the starting point and the core. Gut microbial metabolism complements host metabolism in maintaining metabolic homeostasis of many biologically important endogenous molecules and the disposition of numerous exogenous compounds. Among them, the widely distributed gut bacterial β-glucuronidases (BGUSs) coordinate with host UDP-glucuronosyltransferases (UGTs) to play a role in the occurrence and intervention of diseases by affecting the glucuronidation homeostasis and altering the intestinal local and/or systemic exposure of endogenous compounds and xenobiotics. On one hand, many ingredients of CHMs undergo enterohepatic circulation; On the other hand, CHMs can act on BGUSs directly or indirectly change the distribution and function of BGUSs through reprogramming gut microbiome. The multiple interactions between BGUSs and CHMs may play an important role in the overall therapeutic benefits of CHMs. This work firstly summarizes the latest research progress on BGUSs; then the physiological, pathological and pharmacological significance of BGUSs are exemplified with representative endogenous and exogenous compounds from the aspects of nutrient utilization, metabolic homeostasis, and therapeutic response based on the varied substrate spectra of BGUSs; finally, the scattered data in literature were integrated to summarize the multiple interactions between BGUSs and CHMs, highlighting the important role of BGUSs in the holistic actions of CHMs.

Objective:To investigate the correlation between variations in mitochondrial DNA (mtDNA) control region (D-loop) and keloids.Methods:A total of 216 patients with keloids were collected from Department of Dermatology, the First Affiliated Hospital of Kunming Medical University from 2016 to 2019. Total DNA was extracted from peripheral blood samples of all the patients, as well as keloid tissues and perilesional normal skin tissues of 25 patients with keloids. Peripheral blood samples were collected from 299 health checkup examinees without keloids in Health Examination Center, the Affiliated Hospital of Yunnan University, who served as controls. PCR amplification and Sanger sequencing were performed on the mtDNA D-loop region, and mutation sites in each sample were analyzed by comparisons with the revised Cambridge Reference Sequence (rCRS) . Haplogroups were assigned in the 2 groups by using Phylotree-mtDNA tree Build 17. Mutations in the mtDNA D-loop region were compared among keloid tissues, perilesional normal skin tissues and peripheral blood samples. A median-joining network was constructed via network 5.0 software. Binary logistic regression analysis was performed to investigate the correlation between haplogroup frequencies and the occurrence of keloids, and chi-square, t and t′ tests were used to analyze clinical data. Results:Among the 216 patients with keloids, variations in mtDNA D-loop region were classified into 10 haplogroups, including A, B, D, R9, G, M*, M7, M8, M9 and N9, with the haplogroups R9 and M9 showing the highest (21.3%, 46/216) and lowest (0.9%, 2/216) frequencies respectively. The frequencies of haplogroups M7 ( P=0.040, OR=0.248, 95% CI: 0.066 - 0.937) and N9 ( P=0.048, OR=0.191, 95% CI: 0.037-0.986) were significantly lower in the patients with keloids than in the controls. The median-joining network plot showed that the distribution pattern of the haplogroup M7 differed between the patients with keloids and controls. Significantly less number of lesional sites and younger age of onset were observed in the patients with haplogroup M7 compared with those with non-M7 haplogroups ( P=0.000 1, 0.045, respectively) . Conclusion:The haplogroup M7 is correlated with the occurrence of keloids, and may be a potential protective factor for keloid formation.