Objective To observe the expression of acid-sensing ion channel la (ASICla) and to investigate the effect of intracellular Ca2 + concentration on focal cerebral ischemia in diabetic rats.Methods 108 male Wistar rats were divided into three groups:group A [rats with middle cerebral artery occlusion (MCAO)],group B [rats with MCAO and diabetes (DM + MCAO)],group C [rats with MCAO and diabetes treated with fasudil intervention (DM+ MCAO+ fasudil)] (n=36 each).Samples were obtained at the time points of 1,3,6 and 24 h after ischemia respectively (n=9).Models of MCAO and DM+MCAO were prepared.Rats in DM+MCAO+Fasudil group were treated with fasudil 1 mg/Kg by caudal vein injection after half an hour when DM+MCAO model successfully prepared.ASICla expressions were detected at different time points of ischemia in the 3 groups respectively.Ca2+ concentration in ischemia cortex cells were determined at different time points of ischemia in group B and C.Results ASICla expressions were gradually increased along with the ischemia time in group A and B (group A:0.71±0.10,0.80±0.11,0.86±0.08,0.93±0.09;groupB:0.86±0.11,1.05±0.51,2.42±0.08,2.78±0.04; all P＜ 0.05),and ASICla expressions at different time points were higher in group B than in group A (all P＜ 0.05).Ca2-concentration were gradually increased along with the ischemia time in group B (106.32± 18.6,137.84±14.32,151.94± 18.38,183.61±7.96,all P＜0.05).Compared with group B,the levels of ASICla expression and calcium current were reduced in group C.Conclusions The activation of ASICla increases calcium ion flow internal pathway leading to intracellular calcium overload,which may be one of the reasons for the aggravation of focal cerebral ischemia in diabetes.

Objective:To explore whether microRNA-21-5p (miR-21-5p) has the effect of anti-apoptosis of human alveolar typeⅡ epithelial cells (ATⅡ).Methods:ATⅡ cells derived from the human were cultured in vitro and used for experiments when the cells were grown until the presence of lamellar bodies and microvilli were observed by light microscope. The cells were divided into blank control group (direct culture), hydrogen peroxide (H 2O 2) injury group (cultured with 0.5 mmol/L H 2O 2), and miR-21-5p overexpression group (using miR-21-5p with a multiplicity of infection (MOI) of 100 lentiviral overexpression vector with 0.5 mmol/L H 2O 2) and miR-21-5p empty virus control group (miR-21-5p lentiviral blank vector was co-cultured with 0.5 mmol/L H 2O 2). In each group, cell proliferation was detected by cell counting kit-8 (CCK-8) at 0, 12, 24, 36, and 48 hours of cell culture; cell apoptosis was detected by flow cytometry at 24 hours of culture. Results:① Cell proliferation activity test results: with the extension of cell culture time, the cell proliferation activity of the blank control group gradually increased, while the cell proliferation activity gradually decreased after the addition of 0.5 mmol/L H 2O 2. However, the cells proliferation activity in the miR-21-5p overexpression group decreased more slowly than that in the H 2O 2 injury group and the miR-21-5p empty virus control group, and the cell proliferation activity at 48 hours was significantly higher than the H 2O 2 injury group and the miR-21-5pempty virus control group ( A value: 0.295±0.005 vs. 0.184±0.005, 0.169±0.002, both P < 0.05). It showed that both H 2O 2 and lentivirus accelerated cell damage, while miR-21-5p could reduce cell apoptosis. ② Apoptosis rate test results: compared with the blank control group, the apoptosis rate increased significantly after adding 0.5 mmol/L H 2O 2; while the apoptosis rate of the miR-21-5p overexpression group was lower than that of the H 2O 2 injury group and miR-21-5p empty virus control group [early apoptosis rate: (14.31±0.12)% vs. (24.50±0.12)%, (23.41±0.13)%; late apoptosis rate: (8.12±0.13)% vs. (9.71±0.11)%, (10.41±0.15)%; overall apoptosis rate: (22.33±0.12)% vs. (34.21±0.10)%, (33.82±0.14)%; all P < 0.05], which further proved that miR-21-5p had anti-apoptotic effects. Conclusion:miR-21-5p has an anti-apoptotic effect on human ATⅡ.

Objective To explore the mutation and evolution of Yersiniapestis(Y. pestis) from the point of codon and 16S-ribosome. Methods Codon preference and 16S-ribosome of Y. pestis were analyzed by bioinformatics. Results Similar codon preference was found among 4 PCD1 Y. pestis, of the 3 old Y. pestis the codon preference between PMT1 and PCD1 was similar. There were some differences between PCD1, PCP1 and Yunnan 6 kb plasmid. Through the analysis of 16S-ribosome, the sequences were found similar in 11 strains of Y. pestis,Yersinia pseudotuberculosis was very close to Y. pestis, with only one nucleotide difference, mutated G-T, and corresponding amino acid methionine (M)-isoleucine (I). There were some differences in sequences of 16S-ribosome in Y. pestis, Escherichia coli and Pulex irritans. Conclusions The time for Y. pestis to obtain PCP1 is later than PMT1 does, in other words, the affinity of Y. pestis with PMT1 was closer than PCP1 with 6 kb plasmid;alteration of 16S-ribosomal nucleotide sites may cause changes in function and structure of 16S rRNA. The lower similarity between 16S-ribosomal sequences of Y. pestis and Pulex irritans indicates the time for co-evolution is very short,and the late emergence of Y. pestis.

Objective To study the effects of exogenous melatonin on plasma corticosterone(CS)and adrenal glucocorticoid receptor(GR)mRNA in neonatal rats after hypoxic-ischemic brain damage(HIBD). Methods One hundred and forteen 7-day-old Sprague-Dawley rats were randomly divided into four groups:model group(K group),sham operation group(H group),melatonin-treated group(T group),and normal control group(S group). Blood was obtained in different time points after HIBD. Plasma CS levels were measured by radioimmunoassay(RIA). The expression of GR mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR). Results Plasma level of CS increased and the expression of GR mRNA decreased significantly after HIBD,however,these changes were markedly reversed by exogenous melatonin. Conclusions Plasma CS and adrenal gland GR were involved in the pathological process of HIBD. Exogenous melatonin could up-regulate the expression of GR by decreasing the level of endogenous CS,so it may alleviate the excess stress injury after HIBD.

Objective To summarize the nursing points for patients with primary hepatocellular carcinoma (HCC) who are receiving CT-guided percutaneous microwave ablation treatment. Methods A total of 77 HCC patients were enrolled in this study. CT-guided percutaneous microwave ablation treatment was carried out in all patients. High quality perioperative nursing care was implemented, and the adverse reactions were analyzed and summarized. Results CT-guided percutaneous microwave ablation was successfully accomplished in all the 77 cases. After the treatment, different degrees of pain occurred in 29 cases, liver function damage was detected in 6 cases, and pneumothorax was seen in one case. After active treatment and nursing, the clinical conditions were effectively controlled in all patients. No death occurred in perioperative period. Conclusion With the help of sufficient preoperative preparation, close intraoperative cooperation and postoperative effective treatment and nursing, the clinical condition is significantly improved, the complications have been effectively prevented and reduced, and the patient’s quality of life is also improved.

OBJECTIVEOur previous work indicated that overexpression of imprinting gene PEG10 is associated with malignant phenotype of hepatocellular carcinoma. The aim of this study is to explore whether disregulation of PEG10 leads to dysregulation of microRNAs.

METHODSIn silico analysis using TargetScan indicated that miR-122 could regulate the expression of PEG10. The expression of miR-122 in three hepatoma cell lines, Huh7, Hep3B and HepG2 and in primary human normal liver cell were compared using real time RT-PCR. After pre-miR-122 was transfected into HepG2 cell, the levels of PEG10 mRNA and protein were measured.

RESULTSIn silico analysis revealed that miR-122 could regulate the expression of PEG10. Real time RT-PCR indicated that miR-122 was not expressed in Hep3B and HepG2 cells, and only weakly expressed in Huh7 cells, but highly expressed in primary human normal liver cells. The expression of miR-122 was negatively correlated with the expression of PEG10. After pre-miR122 was transfected into HepG2, the mRNA level of PEG10 was not increased, whereas the protein level of PEG10 was increased.

CONCLUSIONmiR-122 may be involved in regulation of PEG10 expression.

Carcinoma, Hepatocellular ; genetics ; Gene Expression Regulation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; MicroRNAs ; genetics ; Proteins ; metabolism

OBJECTIVETo explore the mechanism by which HBV X gene(HBx) inhibits apoptosis of human hepatoma cell line HepG2 in terms of miRNA.

METHODSThree cell lines were prepared: HepG2 cells stably transfected with HBx (HepG2/HBx), HepG2 cells stably transfected with pcDNA3.1 (HepG2/pcDNA3.1) and HepG2 cells. Flow cytometry was adopted to measure the apoptosis of these three cells and Taqman fluorescence quantitative PCR was used to examine miR-192 expression. After HepG2 cells was transfected with miR-192, the apoptosis was analyzed by flow cytometry and the expressions of p53 and PUMA at mRNA and protein levels were evaluated by SYBR Green quantitative PCR and Western blot, respectively.

RESULTSCompared with HepG2/pcDNA3.1 cells (11.46% ± 0.69%) and HepG2 cells (12.5% ± 0.66%), the apoptosis rate of HepG2/HBx cells (2.37% ± 0.35%) was significantly reduced (F = 171.722, P < 0.01). The level of miR-192 was 49.1% ± 5.9% in HepG2 cells, which was dramatically down-regulated (F = 14.319, P = 0.019) as compared to the other two groups (HepG2/pcDNA3.1: 98.0% ± 8.9%; HepG2: 100%). Compared with HepG2 cells transfected with miR-NC (10.74% ± 1.15%), transfection of miR-192 into HepG2 cells led to increased apoptosis (15.74% ± 1.17%) (F = 18.415, P = 0.013) and higher p53 and PUMA expressions at mRNA (p53: 1.68 ± 0.12 vs 0.90 ± 0.09, F = 43.115, P = 0.003, PUMA: 1.66 ± 0.10 vs 0.98 ± 0.06, F = 22.541, P = 0.009) and protein (p53: 3.07 vs 1, PUMA: 2.13 vs 1) levels.

CONCLUSIONHBx could inhibit apoptosis of HepG2 cells through down-regulation of miR-192 which induces apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Down-Regulation ; Genes, Viral ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; metabolism ; Trans-Activators ; genetics ; metabolism

[Abstra ct] Objective To investigate the long-term efficacy and adverse effects of intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC).Methods A total of 869 patients with biopsy-proven NPC without distant metastasis who underwent the whole course of IMRT from 2009 to 2010 were enrolled.Of all the patients, 84.8%received cisplatin-based chemotherapy.The prescribed dose to the primary lesion in the nasopharynx was 66-70Gy in 30-32 fractions, and the dose to the positive lymph nodes in the neck was 66 Gy in 30-32 fractions.The Kaplan-Meier method was used to calculate survival rates, the log-rank test was used for difference analysis and univariate prognostic analysis , and the Cox proportional hazards model was used for multivariate prognostic analysis .Rseu lts The 5-year overall survival( OS ) , local recurrence-free survival, regional recurrence-free survival, distant metastasis-free survival, and disease-free survival ( DFS ) were 84.0%, 89.7%, 94.5%, 85.6%, and 76.3%, respectively.In the patients with locally advanced NPC,concurrent chemotherapy tended to reduce distant metastasis (83.6%vs.75.7%, P=0.050) and improve OS (82.6%vs.77.0 %, P=0.082).Induction chemotherapy tended to improve OS ( 80.7% vs.71.4%, P=0.057 ) , and the induction chemotherapy containing docetaxel or gemcitabine tended to improve OS (83.3%vs.72.2%, P=0.058).The patients who received a boost after the initial radiotherapy had a significantly lower DFS rate than those who did not (52.2%vs.71.1%, P=0.004).The concurrent chemotherapy increased the incidence rates of long-term xerostomia and trismus, while a high dose of cisplatin increased the incidence rates of xerostomia and hearing impairment.Conclusions IMRT for NPC provides satisfactory long-term efficacy.Concurrent chemotherapy combined with IMRT tends to reduce the incidence of distant metastasis, and other values need further investigation.The boost therapy after radiotherapy may be associated with poor prognosis.Chemotherapy increases the incidence of long-term toxicities.

The present study investigated the cellular mechanism underlying the effect of ligustrazine on the ion transport in rat distal colon using the short-circuit current (I(SC)) technique. In freshly isolated colonic strips, basolateral addition of ligustrazine stimulated a rise in I(SC), which was resistant to basolateral application of neuronal sodium channel blocker tetrodotoxin (TTX), but inhibited by 55.2% by basolateral pretreatment with prostaglandin inhibitor indomethacin. The ligustrazine-induced I(SC) increase was inhibited by apical application of Cl(-) channel blockers diphenylamine-2,2'-dicarboxylic acid (DPC) and glibenclamide. Basolaterally administered bumetanide, an inhibitor of Na(+)-K(+)-2 Cl(-) cotransporter, inhibited ligustrazine-evoked current increases by 85.2% and basolateral exposure to Ba(2+), a non-specific potassium channels blocker, and blocked the current by more than 90%, indicating that basolateral Na(+)-K(+)-2 Cl(-) cotransporter and K(+) channels played an important role in the effect of ligustrazine. The results suggested that ligustrazine could stimulate rat distal colon (-) secretion that is mediated by basolateral Na(+)-K(+)-2 Cl(-) cotransporter and K(+) channel.
Animals ; Animals, Newborn ; Calcium Channel Blockers ; pharmacology ; Colon ; metabolism ; physiology ; Epithelial Cells ; metabolism ; Evoked Potentials ; drug effects ; In Vitro Techniques ; Intestinal Mucosa ; cytology ; metabolism ; Ion Transport ; drug effects ; Male ; Potassium Channels ; metabolism ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE To evaluate the feasibility and sterile effect of low-temperature plasma sterilizer produced by Chinese Academy of Sciences for the sterilization of arthroscope surgical instruments.METHODS To explore the cleaning,package,arrangement and sterile effect monitoring of instruments in the process of arthroscope sterilization using low-temperature plasma sterilizer.RESULTS In the sterilization process of arthroscope surgical instruments using low-temperature plasma sterilizer,the chemical indicators and indicating patch were discolored,the monitoring of bio-box and the sterilization monitoring of surgical instruments were all negative.CONCLUSIONS Using the low-temperature plasma sterilizer machine for the sterilization of endoscope surgical instruments can ensure the security of sterilization,make less injury of precise instruments,shorten the sterilization time,accelerate the surgical turnover,enhance the frequency of surgical instruments application,and increase the efficiency.