OBJECTIVE: Investigate the effect of oxidative stress on the occurrence and development of laryngeal squamous cell carcinoma associated with smoking, and the clinical diagnostic value of catalase on smoking related laryngeal squamous cell carcinoma. METHOD: Collecting 119 smokers(including the smoking related laryngeal cancer group 68 cases, the control group 51 cases), the indexes of catalase (CAT), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH), nitric oxide (NO) in blood plasma and cancerous tissue in two groups were compared. The association between these oxidative stress indicators and the occurrence and severity of smoking related laryngeal squamous cell carcinoma was analysised by SPSS 17.0. RESULT: (1) Compared with control group, the smoke frequency and amount, CAT, MDA, GSH increased significantly in the smoking related laryngeal cancer group (P = 0.000; 0.000; 0.000; 0.000; 0.000); whereas SOD, NO decreased (P = 0.000; 0.000). (2) The lower the differentiation degree, the higher the serum CAT (P = 0.000) and the higher CAT, MDA, GSH of larynx tissue (P = 0.000; 0.000; 0.000), but the lower the serum NO (P = 0.000) and the lower SOD, NO of larynx tissue (P = 0.000; 0.000); The higher the clinical stage, the higher CAT of serum and larynx tissue and the higher GSH of larynx tissue (P = 0.000; 0.001), the lower NO of larynx tissue (P = 0.009). (3) The serum CAT, MDA were independent risk factors of smoking related laryngeal squamous cell carcinoma (OR = 1.060, 2.475; P < 0.01, P < 0.05). CONCLUSION Oxidative stress is the key factor of the occurrence of smoking related laryngeal squamous cell carcinoma, and the CAT can be used as the indicator of clinical diagnosis of smoking related laryngeal squamous cell carcinoma.
Antioxidants ; metabolism ; Carcinoma, Squamous Cell ; enzymology ; Catalase ; metabolism ; Glutathione ; metabolism ; Head and Neck Neoplasms ; enzymology ; Humans ; Laryngeal Neoplasms ; enzymology ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Oxidative Stress ; Risk Factors ; Smoking ; adverse effects ; Squamous Cell Carcinoma of Head and Neck ; Superoxide Dismutase ; metabolism

OBJECTIVETo evaluate interfering effect of several short interfering RNAs (siRNA) on HCV 5' untranslated region(5' UTR).

METHODSThe green fluorescent protein (GFP) was used as reporter gene. A fused gene of HCV-5'UTR and GFP was constructed. It was cloned into the plasmid pCDNA3.1 named as pcDNA-HCV-5'UTR-GFP. Three siRNAs were designed and transfected into HepG2 cells with pcDNA-HCV-5'UTR-GFP. The change of the fluorescence intensity of HepG2 cells was shown by fluorescence microscopy and numerically detected under 488 nm wave length by flow cytometry.

RESULTSThe fused gene of HCV-5'UTR and GFP was successfully constructed. The seven groups displayed inhibitory effects on the gene expression of GFP. The inhibition rates of siRNA A, B and C were 68.4 percent, 72.6 percent and 75.6 percent, respectively. The inhibitory rates of siRN A + B, siRN B +C and siRN A +C were 91.8 percent, 87.2 percent and 92.4 percent, respectively. The inhibitory rates of siRN A+B +C was the highest, up to 95.7 percent.

CONCLUSIONThese siRNAs could inhibit expression of HCV 5'UTR gene, the inhibitory effect of combined siRNA was better than that of single siRNAs.

5' Untranslated Regions ; genetics ; Green Fluorescent Proteins ; genetics ; Hepacivirus ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

This paper is to report the polymorphism of raw materials of clopidogrel bisulfate at home and abroad. By the analysis of Fourier transform infrared spectroscopy (FTIR) and powder X-ray diffraction (p-XRD), samples are roughly classified into two groups, except one patent material. And the differential scanning calorimeter (DSC) examination showed more detailed information for these materials. The results of the study could provide comprehensive basis for the quality evaluation of clopidogrel bisulfate.
Calorimetry, Differential Scanning ; Crystallization ; Evaluation Studies as Topic ; Platelet Aggregation Inhibitors ; chemistry ; Spectroscopy, Fourier Transform Infrared ; Ticlopidine ; analogs & derivatives ; chemistry ; X-Ray Diffraction

Objective To investigate the correlation between NRAMP1 gene polymorphisms and susceptibility to tuberculosis ( TB) in Tibetan people in Qinghai. Methods A case-control study was con-ducted in this study, involving 99 Tibetan patients with TB and 89 healthy Tibetans. The single nucleotide polymorphisms of NRAMP1 gene at rs17235409 and rs3731865 sites were detected by using TaqMan probe method. Gene cloning and sequencing typing were performed to analyze the single nucleotide polymorphisms of NRAMP1 gene at the rs17235416 site. SPASS20. 0 software was used to statistically analyze the correla-tion between NRAMP1 gene polymorphisms and susceptibility to TB in Tibetan people. Results No signifi-cant difference in the genotype frequencies of rs3731865 and rs17235409 was found between the two groups (χ2=0. 852, P=0. 356;χ2=0. 279, P=0. 597). The genotype frequencies of TGTG/TGTG and TGTG/del+del/del at the rs17235416 site were 70. 7% ( 70/99 ) and 29. 3% ( 29/99 ) in patients with TB and 86. 5% (77/89) and 13. 5% (12/89) in healthy subjects. There were significant differences in the geno-type frequencies of TGTG/TGTG and TGTG/del+del/del between the two groups (χ2=6. 870, P=0. 009). The genotypes of TGTG/del and del/del at rs17235416 were risk factors for TB ( OR=0. 376; 95%CI:0. 178-1. 794 as compared with the TGTG/TGTG genotype in Tibetan people in Qinghai. Conclusion This study suggested that the NRAMP1 gene polymorphisms at rs3731865 and rs1723409 sites had no correlation with the susceptibility to TB in Tibetans in Qinghai. However, the NRAMP1 gene polymorphisms at rs17235416 site were correlated with the susceptibility to TB. The TGTG/del alleles at the rs17235416 site might be the risk factors for tuberculosis in Tibetans in Qinghai.

Objective To investigate the function of autophagy in the process of PM2.5-induced apoptosis. Methods PM2.5 was obtained from Zhanjiang in 2016. Human lung adenocarcinoma cells H441 were treated with PM2.5 at different concentrations for different times. Cell proliferation was analyzed by MTT assay; Cell apoptosis was assessed by PI and Annexin V double staining and TUNEL assay. The expression of autophagy marker LC3Ⅱ, AKT and P-AKT protein was examined by Western blot ( WB). H441 cells were treated with PM2.5 following treatment with rapamycin or 3-MA. Cell viability was evaluated by trypan blue staining. Results Compared with the control group, cell proliferation was significantly inhibited by PM2.5 at concentration of 100 μg/mL or more for 24 and 48 h. With the increase of PM2.5 concentration, the cells apoptotic rate significantly increased, the protein ex-pression of LC3Ⅱwas increased as well as the P-AKT was decreased; and the protein expression of LC3Ⅱwas in-creased significantly after AKT inhibitor treatment. Moreover, rapamycin decreased PM2.5-induced cell apoptosis, and 3-MA can promote PM2.5-induced cell apoptosis. Conclusions In H441 cells, PM2.5 activates autophagy by inhibiting activation of AKT pathway, and cell autophagy can mitigate PM2.5-induced apoptosis.

Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the adult central nervous system (CNS), however, it causes excitation in the immature CNS neurons. The shift from GABA-induced depolarization to hyperpolarization in postnatal brain is primarily due to progressive decrease in the expression of the Na-K-2Cl symporter 1 (NKCC1) and increased expression of the K-Cl cotransporter 2 (KCC2). Unlike CNS neurons, both immature and mature neurons in the enteric nervous system (ENS) are depolarized by GABA. Molecular mechanisms by which GABA excites ENS neurons are unclear. It is understood, however, that the excitatory action depends on elevated intraneuronal Cl. We aimed to test a hypothesis that high intracellular Cl in ENS neurons is maintained by activity of the NKCCs. We found that NKCC2 immunoreactivity (IR) was expressed in the ENS of the rat colon on postnatal day 1 (P1). The expression level of NKCC2 continuously increased and reached a steady high level on P14 and maintained at that level in adulthood. NKCC1 IR appeared in ENS on P14 and maintained through adulthood. KCC2 IR was not detectable in the ENS in any of the developmental stages. Both NKCC1 IR and NKCC2 IR were co-expressed with GABA receptors in ENS neurons. Exogenous GABA (1 mmol/L) caused membrane depolarization in the ENS neurons. The reversal potential of GABA-induced depolarization was about -16 mV. Blockade of NKCC by bumetanide (50 μmol/L) or furosemide (300 μmol/L) suppressed the depolarizing responses to GABA. Bumetanide (50 μmol/L) shifted the reversal potential of GABA-induced depolarization in the hyperpolarizing direction. Neither the KCC blocker DIOA (20 μmol/L) nor the Cl/HCO exchanger inhibitor DIDS (200 μmol/L) suppressed GABA-evoked depolarization. The results suggest that ENS neurons continuously express NKCC2 since P1 and NKCC1 since P14, which contribute to the accumulation of Cl in ENS neurons and GABA-evoked depolarization in neonate and adult ENS neurons. These results provide the first direct evidence for the contribution of both NKCC2 and NKCC1 to the GABA-mediated depolarization.
Animals ; Bumetanide ; Neurons ; Rats ; Receptors, GABA-A ; Symporters ; gamma-Aminobutyric Acid

OBJECTIVETo screen for genetic mutations in 35 patients with Leber's hereditary optic neuropathy (LHON).

METHODSPolymerase chain reaction and DNA sequencing were used to screen for the presence of mitochondrial DNA mutations.

RESULTSThe total detection rate of top 3 common LHON mutations were 20.0%, which included 6 cases of ND4 11778 G to A, 1 case of ND1 3460 G to A. No ND6 14484 T to C mutation was detected. A ND4 G11719A synonymous mutation was found in all patients. In addition, 21 other mutations were discovered among 23 patients, among which 13 had a single mutation, 8 had a second mutations, and 2 had a third mutation. Among the 21 mutations, ND4 11778 G to A had a frequency of 28.6%(6/21). ND1 3552 T to A, ND6 14470 T to C, ND4 11794 T to C, ND1 3497 C to T and 3644 T to C respectively had a frequency of 19.0% (4/21), 19.0%(4/21), 14.3%(3/21), 9.5%(2/21) and 9.5%(2/21). Among the 3 patients who harbored a ND4 11794 T to C mutation, 2 were heteroplasmic and one was homoplasmic in nature.

CONCLUSIONThe ND4 11778 G to A mutation is common in the Top "3" primary mutations of patients with LHON. Candidate LHON mutation ND1 3552 T to A or ND1 3644 T to C resulted in LHON pathogenesis as single or synergistic effect. The visual impairment at onset of the disease with candidate mutation were better than the eyes with the ND4 11778 G to A mutation.

Adolescent ; Adult ; Child ; Child, Preschool ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Optic Atrophy, Hereditary, Leber ; genetics

To study the new antifungal active triterpene glycosides of sea cucumber Holothuria scabra. Triterpene glycosides from Holothuria scabra were separated and purified by silica gel chromatography, reversed-phase silica gel chromatography and RP-HPLC. Their structures were elucidated on the basis of spectral data and chemical evidence. Three triterpene glycosides were identified as scabraside A (1), echinoidea A (2) and holothurin A1 (3). Scabraside A (1) is a new triterpene glycoside, and compounds 2 and 3 were isolated from Holothuria scabra for the first time. They showed antifungal activities (1 < or = MIC80 < or = 16 microg mL(-1)).
Animals ; Antifungal Agents ; isolation & purification ; pharmacology ; Glycosides ; chemistry ; isolation & purification ; pharmacology ; Holothuria ; chemistry ; Holothurin ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

BACKGROUNDThis study aimed to evaluate the effects of standard rescue procedure (SRP) in improving severe trauma treatments in China.

METHODSThis study was conducted in 12 hospitals located in geographically and industrially different cities in China. A standard procedure on severe trauma rescue was established as a general rule for staff training and patient treatment. A regional network (system) efficiently integrating prehospital rescue, emergency room treatments, and hospital specialist treatments was built under the rule for information sharing and improving severe trauma treatments. Treatment outcomes were compared between before and 1 year after the implementation of the SRP.

RESULTSThe outcomes of a total of 74,615 and 12,051 trauma cases were collected from 12 hospitals before and after the implementation of the SRP. Implementation of the SRP led to efficient cooperation and information sharing of different treatment services. The emergency response time, prehospital transit time, emergency rescue time, consultation call time, and mortality rate of patients were 24.24 ± 4.32 min, 45.69 ± 3.89 min, 6.38 ± 1.05 min, 17.53 ± 0.72 min, and 33.82% ± 3.87% (n = 441), respectively, before the implementation of the standardization and significantly reduced to 10.11 ± 3.21 min, 22.39 ± 4.32 min, 3.26 ± 0.89 min, 3.45 ± 0.45 min, and 20.49% ± 3.11%, separately (n = 495, P < 0.05) after that.

CONCLUSIONSStaff training and SRP can significantly improve the efficiency of severe trauma treatments in China.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Emergency Medical Services ; standards ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Wounds and Injuries ; Young Adult

Objective:To establish a molecular identification method for Bupleurum chinense seeds based on ribosomal DNA internal transcribed spacer (ITS) sequence, ensuring the species authenticity of the cultivated seeds of B. chinense. Method:A total of 59 seeds samples of B. chinense and its main cultivated species, marketed B. chinense were collected. The effect of different sampling amounts and different water bath conditions on DNA extraction quality of the seeds was investigated, a DNA extraction method for seeds of Bupleurum was established. Their ITS sequences were obtained by polymerase chain reaction (PCR) and bidirectional sequencing. In addition, 34 ITS sequences of main cultivated Bupleurum species, such as B. chinense, B. scorzonerifolium, B. falcatum and B. smithii, were downloaded from GenBank to enrich identification database of B. chinense seeds. The neighbor-joining (NJ) dendrogram were constructed by MEGA-X 10.0.5 software to investigate the the species identification ability of ITS sequences for B. chinense seeds. And DNA barcoding identification of marketed B. chinense seeds was conducted based on BLAST method and NJ dendrogram method. Result:In total, 59 ITS sequences were obtained. ITS sequences of B. chinense could be divided into six haplotypes, including seven variable sites. The NJ dendrogram indicated that all the haplotypes of B. chinense could form independent branches, which could be distinguished from other cultivated species of Bupleurum in the collected samples, and possessed the ability to identify species of B. chinense seeds. Based on ITS sequence barcoding identification, 3 of the 19 marketed B. chinense seeds were B. falcatum with a counterfeit rate of 15.8%. Conclusion:DNA barcoding technology based on ITS sequence can accurately and reliably identify B. chinense seeds and its adulterants, providing reference for the standardization construction of Chinese medicinal materials seeds.