(3S)-Linalool synthase (LIS) is a key enzyme involved in the monoterpene biosynthetic pathway. Based on our previous transcriptome study, the expression level of LIS gene was exceedingly related to glycyrrhizic acid (GA) biosynthesis. Therefore, we used hairy root culturing to further investigate the effect of LIS on the GA biosynthesis. A LIS gene (GenBank accession number: MZ169552) was cloned from Glycyrrhiza uralensis. The plant binary overexpression vector pCA-LIS was constructed by gene fusion. G. uralensis hairy roots overexpressing LIS were induced by the Agrobacterium rhizogenes ATCC15834. The expression levels of LIS were analyzed by real-time quantitative PCR (RT-qPCR) and the contents of GA in hairy root lines were determined by UPLC. It was found that in the hairy root lines overexpressing LIS, the expression levels of LIS were significantly higher than that in the wild type, while the contents of GA were remarkably lower than those in the wild type and negative control. These findings indicate that the expression level of LIS is negatively correlated with the accumulation of GA. In this study, LIS was cloned from G. uralensis for the first time and the negative regulatory effect of LIS on GA biosynthesis was confirmed by reverse genetics. This work provides support for further improvement of the molecular regulatory network of GA biosynthesis in G. uralensis.

OBJECTIVE: To study the correlation between ultrastructure of lymphatic capillary in pericancerous tissue and neck lymph node metastasis in laryngeal carcinoma. METHOD: Transmission electronic microscope was applied to observe and compare ultrastructure of lymphatic capillary in 8 normal laryngeal epithelial tissue and 12 pericancerous tissue of laryngeal cancer. RESULT: Lymphatic capillary of pericancerous tissue was significantly dilated compared with normal laryngeal tissue. A large amount of endothelial cell junction was open and devoid of basement membrane. Some endothelial cells of lymphatic capillary were destroyed or broken completely. CONCLUSION Ultrastructure of lymphatic capillary in pericancerous tissue is an important factor of laryngeal cancer metastasis through lymphatic system. This research offers theoretic basis for laryngeal carcinoma metastasis mechanism and prevention.
Aged ; Carcinoma, Squamous Cell ; pathology ; Case-Control Studies ; Female ; Humans ; Laryngeal Mucosa ; pathology ; Laryngeal Neoplasms ; pathology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; Neck ; Neoplasm Staging

Pentacyclic triterpenoids are a class of widespread natural compounds containing six isoprene structures with a wide range of pharmacological activities, including antibacterial, anti-inflammatory, antiviral, antitumor, immune regulation, etc. The structural modifications of pentacyclic triterpenoid natural products and the drug development have always been a hot research topic. This article reviews the recent progresses in the structural modifications, pharmacological effects, and clinical studies of different kinds of pentacyclic triterpenoid natural products.

Objective:To analyze the relationship between the clinical medical college students' time investment (including study, activities, entertainment and exercise) and general self-efficacy (GSE) in a medical university in Liaoning province, China.Methods:The first-year medical students were asked to participate the survey. Their GSE was measured by using general self-efficacy scale (GSES) in 2018. One year later, the independent variable table was used to investigate the extracurricular activity time, and 683 valid questionnaires were collected. Ordered logistic regression method was used to analyze the correlation between students' extracurricular activities and GSE.Results:Medical students' GSE was positively associated with their time in extracurricular study ( OR = 1.94, 95%CI = 1.49-2.54), volunteer activities ( OR=1.36, 95%CI = 1.01-1.83), and physical activities ( OR = 1.37, 95%CI = 1.01-1.85). However, there was no significant correlation with the time in activities organized by students ( OR = 1.09, 95%CI = 0.79-1.50) or activities organized by school ( OR = 1.15, 95%CI = 0.84-1.59). Furthermore, compared with clinical students of "5+3" year program, the 5-year program clinical students had a stronger correlation between medical students' GSE and the input of extracurricular study time. Conclusion:There is a positive correlation between medical students' GSE and their extracurricular time investment, which indicates that increasing medical students' GSE could be an effective method to improve their extracurricular time investment and eventually improve their comprehensive quality.

Antiepileptic drugs and anti-infective drugs are prescribed at the same time in many cases.The clinically important interactions between them may lead to adverse reaction or failure of treatment.We summarize these clinically important interactions with case reports and mechanism of action.The combinations include macrolides and carbamazepine,carbapenems and valproic acid,ciprofloxacin and carbamazepine,phenytoin,triazole antifungal agents and carbamazepine,phenytoin,and phenobarbital,antituberculosis drugs and phenytoin,diazepam,carbamazepine and oxcarbazepine,anthelmintic drugs and phenytoin,carbamazepine and phenobarbital.While new antiepileptic drugs used in China like oxcarbazepine,lamotrigine,topiramate and levetiracetam have little clinically important interaction with anti-infective drugs.

Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.

[Objective] To describe a case of a rare,novel mutation causing recurrent chorioamniotic membrane separation in a Chinese family with combined next-generation sequencing (NGS) and Sanger sequencing.[Methods] For the affected fetus,potential mutation were detected by the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing.And the prenatal diagnosis were identified by Sanger sequencing.[Results] A frameshifting mutation c.1389_1390delAG (inherited from mother),and a missense mutationc.1006 G ＞ C (inherited from mother) have been identified in the affected fetus (the second pregnancy).The prenatal diagnosis of the third fetus turns out to be a carrier,the mutation was inherited from father.[Conclusions] We describe a novel mutation in gene ZMPSTE24,which was considered with mandibuloacral dysplasia with type B,and that may be the cousecoursecausing of recurrent chorioamniotic membrane separation.This rare mutation constitutes an additional heterogeneous defect causing chorioamniotic membrane separation.And the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing allows high resolution characterization of novel mutions that are not readily detected by present methods.

The direct acting substances of Cuscuta chinensis in vivo were preliminarily identified through the correlation analysis of "metabolites-effect identification" model. The ovariectomized female rats were i.g administered with 95% ethanol extract part, 40% ethanol elution part and n-butanol extract part of Cuscuta chinensis. The serum fingerprints of different parts and times of administration were established by UPLC/Q-TOF-MS. At the same time, serum estradiol (E₂), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were detected. Bivariate correlation analysis and grey correlation analysis were used to screen estrogenic components. The results showed that nine direct acting substances in vivo highly related to estrogen effect were found in the drug containing serum, which were hyperoside, astragalin, methyl quercetin glucuronide, quercetin-diglucuronide, quercetin, apigenin, isoquercitrin, kaempferol glucuronide and kaempferol. We can preliminarily screen out the direct acting substance of estrogen effect of Cuscuta chinensis in vivo based on the research idea of serum spectrum effect correlation. It provides a reliable basis for revealing the estrogeneffective substances of Cuscuta chinensis and confirming the quality markers. This experiment was approved by Harbin University of Commerce Ethics Committees (Approval No. HSDU2020-065).

Objective:To investigate the significance and mechanism of ten-eleven translocation (Tet1) against Mycobacterium marinum ( Mm) infection in mice. Methods:SPF wild-type C57BL/6 and Tet1-knockout (Tet1KO) mice were injected intravenously with Mm. All mice were monitored and the abscesses formed in tail were observed and quantified. Pathological changes in mouse tail tissues were observed using hematoxylin and eosin (HE) staining and transmission electron microscopy and the differences between the two groups were analyzed. Immunohistochemistry staining was used to detect the expression and distribution of TNF-α and TGF-β in mouse tail tissues. Moreover, mouse tail tissues were cultured on 7H10 plates for bacterial counting. The expression of NF-κBp65 and TGF-β was detected by Western blot. Results:Obvious lesions including abscesses and ulcers were formed in the Mm-infected C57BL/6, but only scattered small abscesses were observed in Mm-infected Tet1KO mice. During Mm infection, the bacterial load was gradually increased in C57BL/6 mice, but decreased in Tet1KO mice. Histopathological examination showed that obvious inflammatory cell infiltration and typical granulomatous lesions were found in Mm-infected C57BL/6 mice, while no significant inflammatory cell infiltration was detected in Mm-infected Tet1KO mice. Immunohistochemistry staining demonstrated that the expression of TNF-α and TGF-β was lower in Mm-infected Tet1KO mice than in Mm-infected C57BL/6 mice. Moreover, the expression of phosphorylated NF-κBp65 and TGF-β was significantly reduced in Mm-infected Tet1KO mice as compared with that in Mm-infected C57BL/6 mice. Conclusions:Deletion of Tet1 could alleviate the inflammatory damage mediated by Mm and enhance the host immune response to bacteria.

BACKGROUNDGinsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The present study was undertaken to investigate the effect of ginsenoside Rg3 on human ovarian cancer metastasis and the possible mechanism.

METHODSThe experimental lung metastasis models of ovarian cancer SKOV-3 and the assay of tumor-induced angiogenesis were used to observe the inhibitory effects of Rg3 on tumor metastasis and angiogenesis. The effect of Rg3 on invasive ability of SKOV-3 cells in vitro was detected by Boyden chamber, and immunofluorescence staining was used to recognize the expression of matrix metalloproteinase 9 (MMP-9) in SKOV-3 cells.

RESULTSIn the experimental lung metastasis models of ovarian cancer, the number of tumor colonies in the lung and vessels oriented toward the tumor mass in each ginsenoside Rg3 group, was lower than that of control group. The invasive ability and MMP-9 expression of SKOV-3 cells decreased significantly after treatment with ginsenoside Rg3.

CONCLUSIONSGinsenoside Rg3 can significantly inhibit the metastasis of ovarian cancer. The inhibitory effect is partially due to inhibition of tumor-induced angiogenesis and decrease of invasive ability and MMP-9 expression of SKOV-3 cells.

Animals ; Cell Line, Tumor ; Female ; Ginsenosides ; pharmacology ; Humans ; Lung Neoplasms ; prevention & control ; secondary ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; prevention & control ; Ovarian Neoplasms ; drug therapy ; pathology