OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

AIMTo study the effect of inducible nitric oxide synthase (iNOS) gene mediated by retroviral vector on the proliferation of cultured aortic vascular smooth muscle cell (VSMC) of rat and the possibility of iNOS gene therapy for vessel graft restenosis.

METHODSEx vitro VSMC were transfected by different viral titer of viral supernatant. The expression of the retroviral iNOS transgene was examined by RT-PCR and Western blot analysis. Nitric oxide (NO) release from infected cells was determined by Griess reaction. The inhibition of iNOS transgenosis on the proliferation of VSMC was detected by modified MTT assay.

RESULTSmRNA and protein of transferred iNOS gene were detected 48 hours post-gene transfer within the transfected cells. Levels of iNOSmRNA and protein in PLXSN-iNOS infected cells were positively correlated with viral titer of viral supernatant. PLXSN-treated VSMC showed no evidence of iNOS mRNA and protein. Transfection of PLXSN-iNOS into cultured VSMC resulted in a dose-dependent increase in NO production. And iNOS transgenosis significantly inhibited proliferation of VSMC. The inhibition effect was positively correlated with viral titer of viral supernatant.

CONCLUSIONiNOS gene could be quickly and effectively transferred into cultured VSMC by retroviral vector and its expression could significantly inhibit the proliferation of cultured VSMC. Retrovirus-mediated gene transfer of iNOS might play an important role in prevention of restenosis.

Animals ; Cell Proliferation ; Cells, Cultured ; Genetic Therapy ; Genetic Vectors ; genetics ; Graft Occlusion, Vascular ; prevention & control ; Male ; Myocytes, Smooth Muscle ; cytology ; enzymology ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retroviridae ; genetics ; metabolism ; Transfection

OBJECTIVE: To compare the concentration of teeth DNA extracted by three different pretreatment methods and to explore a simple, economical and practical pretreatment method with high concentration of extracted DNA from teeth. METHODS: A total number of 21 molars were collected from 7 corpses. The pretreatment of 3 molars from each individual was randomly performed by tooth crumb method, ball-milling method and liquid nitrogen milling method and 50 mg tooth crumb was weight and DNA was extracted by AutoMate Express forensic DNA extraction system. Subsequently, the concentration of DNA and corresponding STR genotyping of three methods were compared. RESULTS: The DNA concentration extracted by tooth crumb method, ball-milling method and liquid nitrogen milling method was 0.055 6-1.989 1 ng/μL, 0.036 6-1.175 6 ng/μL and 0.037 8-1.249 0 ng/μL, respectively. The DNA concentration obtained by tooth crumb method was higher (P < 0.05) and the success rate of STR genotyping was high. CONCLUSION Combined with AutoMate Express forensic DNA extraction system, tooth crumb method is an efficient and feasible method to extract DNA from teeth, which can be applied in forensic practice.
DNA/isolation & purification* ; DNA Fingerprinting/methods* ; Genotype ; Humans ; Tooth

RNA has received more attention in the field of forensic medicine and the development of the new biological markers based on RNA shows great significance in the analysis of complex cases. circular RNA (circRNA) is a kind of non-coding RNA which is widely reported recently. Although the regulatory mechanisms of generation and expression are not fully clear, the existing research indicates that circRNA has important biological functions. CircRNA has a cell-type-specific expression with great stability and a high expression level, which makes it meaningful in forensic applications potentially. In this paper, the research progress, the generation and regulation of circRNA as well as its biological characteristics and functions are summarized, which will provide references for related studies and forensic applications.
Forensic Sciences ; Humans ; RNA ; RNA, Circular

Adult ; Aged ; Carcinoma, Small Cell ; pathology ; surgery ; Carcinoma, Squamous Cell ; pathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Thymus Neoplasms ; pathology ; surgery

Functional nucleic acids are natural or artificialnucleic acid sequences with specific functions and special structures. Special metal ions are essential trace elements for human health, but excessive metal ions will be harmful to human health. The functional nucleic acids are widely used in the detection of metal ions because of their advantages such as easy modification, low price, high stability and strong specificity. This review describes the function of functional nucleic acid and metal ions, mainly including cutting type, link type, metal ion-mediated base pairing, click chemical type, conformation change type, and other types. Then, biosensor of functional nucleic acid combined with different signal output is introduced. Finally, the research significance and problems of functional nucleic acid in metal ion detection are discussed, and the future development direction and application prospect of functional nucleic acid biosensor are prospected.

Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.

OBJECTIVETo study the effect of acupuncture on the immune function of patients with colorectal cancer liver metastasis.

METHODSSixty cases with colorectal cancer liver metastasis confirmed by pathology and mageology diagnosis were treated with acupuncture. Zusanli (ST 36), Sanyinjiao (SP 6), Neiguan (PC 6), Shangjuxu (ST 37), Hegu (LI 4), Taixi (KI 3), Taichong (LR 3) ,Yinlingquan (SP 9), Yanglingquan (GB 34), etc. were selected for acupuncture, and Shenque (CV 8), Guanyuan (CV 4), Qihai (CV 6), Zusanli (ST 36) were selected for moxibustion. The changes of CD(3) , CD(4) , CD(8) T cells and NK cells in value were examined with flow cytometry before and after treatment.

RESULTSThe value of T lymphocyte subsets such as CD(3) , CD(4), and CD(8) , as well as NK cells were obviously increased after treatment, and there were significant differences between them before and after treatment.

CONCLUSIONAcupuncture can improve the immune function of patients with colorectal cancer liver metastasis.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; immunology ; pathology ; therapy ; Female ; Humans ; Killer Cells, Natural ; immunology ; Leukocyte Count ; Liver Neoplasms ; immunology ; pathology ; secondary ; therapy ; Male ; Middle Aged ; Neoplasm Metastasis ; T-Lymphocyte Subsets ; immunology

Essential hypertension (EH) is affected by both genetic and environmental factors.The polymorphism of connexin (Cx) genes is found associated with the development of hypertension.However,the association of the polymorphism of Cxs with EH has not been investigated.This study aimed to investigate the association of the polymorphism of connexin (Cx) genes Cx37,Cx40,and Cx43 with EH in Kazak and Han Chinese in Xinjiang,China.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were used to analyze the polymorphism of Cx genes in Kazak and Han EH patients as well as their normotensive controls.The results showed that there were no significant differences in the frequencies of different three genotypes (A/A,A/G,and G/G) and A and G alleles of Cx40 rs35594137 and rs11552588 between EH patients and normotensive controis.However,in Kazak EH patients,the frequencies of three genotypes (A/A,A/G;and G/G) of Cx37 rs1630310 were 24.8％,47.2％ and 28.0％,respectively,which were significantly different from those in Han EH patients.In Han EH patients,the frequencies of the three genotypes (C/C,C/G and G/G) of Cx43 rs1925223 were 6.4％,35.6％ and 58.0％,respectively.Frequencies of the other four genotypes had no statistical differences among Kazak and Hart EH patients and their normotensive controls.These results suggest polymorphisms of Cx37 rs 1630310 and Cx43 rs 1925223 genes may be associated with the pathogenesis of EH.Carrying Cx37 rs1630310-A or Cx43 rs1925223-G genotypes may protect against the development of EH.

OBJECTIVETo evaluate the regional anatomy between the abdominal autonomic nerves including the abdominal aortic plexus (AAP) and the inferior mesenteric artery (IMA), and explore the safe ligation point on the IMA and the optimal dissection method to avoid autonomic nerve injuries.

METHODS AND RESULTSDissections and observation were carried out on 16 fixed male cadavers. The AAP located in the thin fascia layer covering the surface of the aorta and its branches. No autonomic nerves were found in the area around the root of the IMA, and the point where the IMA and the left trunk of the AAP intersected was highly variable. The left trunk of the AAP adhered more closely to the IMA than to the aorta.

CONCLUSIONSIn view of autonomic nerve preservation, the only safe site for ligation of the IMA is at its origin, and no other such sites are available along the IMA trunk and its branches. The IMA and the posterior fascia layer containing the autonomic nerves constitute the optimal surgical plane for IMA ligation, which should be performed following skeletonization of the IMA with careful preservation of the integrity of the posterior fascia layer.

Autonomic Pathways ; anatomy & histology ; surgery ; Cadaver ; Dissection ; methods ; Humans ; Ligation ; adverse effects ; methods ; Mesenteric Artery, Inferior ; surgery ; Preservation, Biological ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Trauma, Nervous System ; etiology ; prevention & control