Objective　To develop a new method to induce the gene transfection in high efficiency for eukaryotic cells in vitro. Methods　 Four kinds of p14ARF gene primarily deleted human carcinoma cell line including H460,A549,U251,and PC-3 were transfected with the human p14ARF expression vector (pCI-neo-p14ARF) by using the new nonliposomal transfection reagent Fugene 6. The efficiency of gene transfer was determined by screening the cells in G418. Results　 After 21 days' selection, G418-resistant clones were shown in all the transfected plate. PCR product of p14ARF gene was positive in all the G418-resistant clones. Cytotoxicity of Fugene 6 was detected. The cell proliferation activity was not affected when it was cultured in a high dose of Fugene 6. Conclusion　 These results demonstrate that Fugene 6 is a rapid, feasible, reproducible, and noncytotoxic gene transfection approach for eukaryotic expression vector in vitro.

Objective: To study quantitative changes of cochlear hair cells and spiral ganglion cells after blast exposure in guinea pigs. Methods: The number of hair cells was calculated using the surface specimen technique and computer image analysis technique after blast exposure. The number of spiral ganglion cells was calculated using pathologic technique and computer image analysis technique after blast exposure. Results: Quantitative observation was carried out 21 days after blast exposure. The number of total hair cells in uncus, the first and second gyre in normal control group and in blast group was 3 599?159.6 and 6 022?98.4 respectively, and the number of spiral ganglion cells in the 2 low parts of cochlear in normal control group and in blast group was 51? 4.72 and 27? 6.94 respectively, the difference in mean value between the groups being significant. Conclusion: Not only the hair cells reduce, but also the spiral ganglion cells are severely damaged after blast exposure. [

Objective To study the expression of Nuclear factor E2 related factor 2 (Nrf2) ,Superoxide Dis‐mutas 1 (SOD1) ,and Heme Oxygenase- 1( HO -1) factors related to Nrf2 signaling pathway in noise-induced cochlear injury of rats .Methods A total of 30 SD rats were randomly and equally assigned into the following 2 groups :noise group and normal control group .The rats in noise group were exposed to an 115 dB SPL continuous steady white noise for 12 hours ,then 1 day ,3 days ,and 7 days later ,the rats receveid the ABR and DPOAE testing after noise exposure .Both RT -PCR and Peggy Sue trace protein detection techniques were used to detect gene ex‐pression in Nrf2 /Keapl-ARE signaling pathway and the protein levels by cochlear tissue sampling after noise ex‐posure 7 days later .Results The ABR thresholds in the noise group was higher than those of in normal control group after noise exposure 1 ,3 ,and 7 days later (P<0 .05) .The DPOAE values of noise group were significantly lower than those of normal control group (P<0 .05) .The up -regulation of expression of Nrf2 ,SOD1 ,HO -1 mRNA(1 .11 ± 0 .05 ,1 .45 ± 0 .12 ,1 .15 ± 0 .03) in noise group were higher than those of in the normal control group after noise exposure 7 days later (1 .00 ± 0 .02 ,1 .10 ± 0 .12 ,0 .92 ± 0 .08) ,and the differences were statistically sig‐nificant (P<0 .05) .And in Peggy Sue trace detection system ,we also found SOD1 ,HO -1 protein levels in noise group were higher than those of in normal control group .Conclusion After the noise -induced cochlear injury in rat ,Nrf2 expression in the cochlea increased and up-regulatd expression of downstream gene and protein content such as antioxidant enzymes SOD1 and HO-1gene by regulating Nrf2 /Keapl-ARE signaling pathway ,which may be involved in protection against oxidative stress injury in noise-induced cochlear injury of rat .

Ca2+ leak via ryanodine receptor type 2 (RyR2) can cause potentially fatal arrhythmias, and RyR2 mutations have been shown in the aetiology of catecholaminergic polymorphic ventricular tachycardia. We report the case of a patient with catecholaminergic polymorphic ventricular tachycardia resulting from a RYR2 mutation who had not only typical electroencephalogram changes, but also epileptiform discharges in electroencephalogram. We believe the changes were closely related to the RYR2 mutation.

Objective To study the effect of ratanasampil (RNSP) which is Traditional Tibetan Medicine on the levels of serum β-amyloid protein, interleukin and tumor necrosis factor alpha (TNF-α) in patients with mild to moderate Alzheimer's disease (AD). Methods One hundred AD patients were divided into two groups in randomized controlled study, including treatment group (RNSP 1 g/d) and control group (piracetam 2.4 g/d). The treatment lasted 12 weeks. The Mini Mental State Examination (MMSE), Alzheimer' s disease Assessment Scale-cognitive subscale (ADAS-cog) and Activity of Daily Living Scale (ADLs) were taken to evaluate the efficacy. Serum levels of amyloid peptides (Aβ40 and Aβ42 ) were measured by ELISA assay. The radioimmunologic assay was used to determine the serum levels of IL-1β, IL-2, IL-6, IL-8 and TNF-α. Results The scores of MMSE, ADAS-cog and ADL significantly improved at 12 weeks after RNSP treatment (P＜0.01, 0.01, 0.05, respectively), while had no significant changes in piracetam group (P＜0.05).The levels of TNF-α, IL-1β, IL-6 and Aβ42 were significantly lower in RNSP group than in Piracetam group (P＜0.01). There was a decrease trend of the Aβ42/Aβ40 ratio at 12 weeks after RNSP treatment (P＜0. 05, P＜0.01 ). The serum Aβ42 level had strong correlations with TNF-α, IL-1 β and IL-6. There were no significant differences in Aβ40 and IL-8 between RNSP group and piracetam group. No obvious drug side effect happened on the groups. Conclusions The reductions of serum TNF-α, IL-1β and IL-6 levels after RNSP treatment may lead to decrease of Aβ42 production in AD patients. RNSP may decrease the Aβ42/Aβ40 ratio and slow down the progress of AD. It may improve the learning and memory ability in treating patients with mild to moderate AD and is well tolerated and safe.

Objective To observe the Klotho expression in kidneys and renal tubular epithelial cells apoptosis in spontaneously hypertensive rats (SHRs) and the effects of cordyceps sinensis (CS), in order to study the mechanism of protective effects of CS on renal tubular cells apoptosis in hypertensive renal damage. Methods Twenty 22-week-old male SHRs were control group. After 8 weeks, the levels of 24 hours urinary protein (Upre), urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (Scr), blood urea nitrogen (BUN) and renal pathological changes were detected; the mRNA expression of Klotho, 053 and 021 was detected by RT-PCR; the protein expression of Klotho, 053, 021 and cleaved-caspase-3 was tested by Western blotting. TUNEL assay was applied to evaluate the renal tubular cell apoptosis. Results As compared to SHR group, the levels of 24 h urinary protein content [(52.16±29.3) mg, (49.97±32.5) mg, (54.67±30.09) mg vs (96.52±36.94) mg], urinary NAG [(44.13±9.11), (42.75±8.33), (41.96±7.88) U/L vs (54.07±6.57) U/L], Sct [(45.25±9.55), (43.76±8.65), (45.18±7.28) μmol/L vs (53.84±10.21) μmol/L]and BUN [(8.25±1.03), (8.40±1.58), (8.32±0.98) mmol/L vs (8.91±1.24) mmol/L]were decreased (all P<0.05), renal pathological changes were relieved, the levels of Klotho expression were up-regulated and the levels of p53 and p21 expression and cleaved-caspase-3 protein expression were down-regulated (all P<0.01), tubular cell apoptosis was decreased [7.56%±0.52%, 7.93%±0.37%, 7.37%±0.36% vs 13.32%±0.64%, P<0.01] in CS, Los and CS+Los group. Conclusions Klotho, p53 and p21 play important roles in renal tubular cells apoptosis in hypertensive renal damage. CS can up-regulate Klotho expression, down-regulate p53 and p21 expression and decrease the cleaved-caspase-3 expression and tubular cell apoptosis to ameliorate the hypertensive renal damage.

OBJECTIVETo compare the clinical effect of the double eyelid blepharoplasty with central minimal incision and with three minimal incisions. Methods: From Jul. 2010 to May 2012, 268 cases (Group A) received double eyelid blepharoplasty with central minimal incision, while 102 cases (Group B) underwent double eyelid blepharoplasty with three minimal incisions. Photos were taken immediately, and 1,2, 4, 8,12 weeks after operation. Operation time, recovery time and postoperative complications were evaluated and recorded. The operation time and recovery time were analyzed by Wilcoxon rank sum test. The postoperative complications were analyzed by chi square test. The satisfactory degree was analyzed by t test.

RESULTSThe operation time in Group A was (25.63 ± 3.74) min, compared with that (29.90 ± 4.13) min in Group B (Z = -8.011, P <0.01). Meanwhile, the recovery time in Group A was shorter than that in Group B (Z = -15.887, P <0.01). The occurrence rate of postoperative complications,including hematoma,recurrence and scar hyperplasia in Group A was also lower than that in Group B. At the same time, the satisfactory degree in Group A was(97.302 ± 1.764), which was higher than that(88.628 10.880) in Group B (t = 12.650, P <0.05).

CONCLUSIONSThe double eyelid blepharoplasty with central minimal incision, which is suitable for all cases except those who has serious blepharochalsais, has more advantages than double eyelid blepharoplasty with three minimal incisions.

Blepharoplasty ; adverse effects ; methods ; Cicatrix ; pathology ; Eyelids ; surgery ; Hematoma ; Humans ; Hyperplasia ; Photography ; Postoperative Complications ; Prospective Studies ; Recurrence

Objective： To study the application of DSA in the diagnosis and treatment of ENT diseases. Methods： The diagnostic and therapeutic roles of DSA in ENT patients admitted from November 1995 to December 1999 were retrospectively studied. Results： Therapeutic vascular embolization using DSA was performed in 9/10 patients with severe epistaxis. The treatment was successful in 8/9 patients with a successful rate of 88.89%; embolization of tumor supplying vessels using DSA as a preoperative measure for reducing operative blood loss in 3 patients with nasopharyngeal fibrohemangioma obtained a total success; diagnosis was clarified in 2 patients using DSA. No patients were with severe complications. Conclusion： DSA is not only a safe and effective measure for diagnosis and therapy, but also effective in differential diagnosis of space occupying lesions. Preoperative selective embolization of tumor supplying arteries can reduce operative blood loss.

OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) extract and losartan (Los) on the expression of Klotho (Kl), P53, P21, and apoptosis in renal tubular epithelial cell NRK-52E induced by angiotensin II (Ang II), and to elucidate its therapeutical mechanism in Ang II induced renal tubular epithelial cell apoptosis. METHODS: NRK-52E cells were incubated with CS with or without Ang II for 24 hours. Experimental groups were divided according to the increasing concentrations of CS:0 (serving as controls), 5, 10, 20, 40, and 80 mg/L. The optimal concentration of CS was selected and cells were divided into 5 groups: controls, Ang II (1*10(-8) mol/L), Ang II (1*10(-8) mol/L)+CS (40 mg/L), Ang II (1*10(-8) mol/L)+Los (1*10(-5) mol/L), and Ang II (1*10(-8) mol/L)+CS (40 mg/L)+Los (1*10(-5) mol/L). After 24 hours, cell proliferation was evaluated by MTT assay. The mRNA and protein expression of Kl, P53 and P21 were measured by RT-PCR. Activity of caspase-3 was evaluated by caspase-3 activity assay Kit. Cell apoptosis was determined by Annexin V-FITC/PI double staining and flow cytometry. RESULTS: Certain concentrations of CS promoted the proliferation of NRK-52E cells and increased cells proliferation inhibited by Ang II (P<0.01 or P<0.05 ). Ang II significantly down-regulated the mRNA and protein expression of Kl, and up-regulated the levels of P53 and P21. Caspase-3 activity and apoptotic rates were decreased, too (all P values<0.01). CS or/and Los significantly increased the expression of Kl mRNA and protein down-regulated by Ang II, decreased P53 mRNA and protein expression, P21 mRNA and protein expression,and inhibited caspase-3 activity and apoptotic rates(all P values<0.05). No cooperative effects were observed in the two drugs (P>0.05). CONCLUSION CS can increase the expression of Kl down-regulated by Ang II, decrease P53 and P21 expression and caspase-3 activity, and reduce Ang II induced NRK-52E cell apoptosis, which may be part of its mechanism of the protective effects on hypertensive renal damage.
Angiotensin II ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Cordyceps ; chemistry ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; metabolism ; Glucuronidase ; genetics ; metabolism ; Humans ; Kidney Tubules ; cytology ; metabolism ; Losartan ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Objective: To analyze the trends and influencing factors of endurance performance of 19-22 years old college students in Hunan Province from 1985 to 2019, so as to provide objective and scientific basis for sports and health work in colleges and universities. Methods: A retrospective analysis was conducted on the data of 14 490 college students aged 19-22 in Hunan Province from 8 consecutive National Student Physical Fitness and Health Surveys conducted from 1985 to 2019. The analysis indexes were 1 000 m running for boys and 800 m running for girls. Results: From 1985 to 2019, the endurance running time of 19-22 years old Han college students in Hunan Province showed an obvious trend of decline. The 1 000 m running time of urban and rural male students increased by 41.9 and 45.4 s on average, and the 800 m running time of urban and rural female students increased by 29.5 and 30.6 s on average, respectively. Multiple linear regression analysis showed that age ( β =0.17), urban students (rural students as reference; β =0.44), GDP ( β =0.94) and urbanization level ( β = 0.44 ) were positively correlated with the average endurance running time of males. Urban students ( β =0.92), GDP ( β = 1.38 ) and Engel coefficient ( β =0.93) were positively correlated with the average endurance running time of females. BMI ( β =-0.47) was negatively correlated with the females mean time of endurance running ( P ＜0.05). Conclusion The endurance performance of Han college students in Hunan Province showed a declining trend from 1985 to 2019,which is associated with age, urban and rural distribution, regional GDP, Engel s coefficient, urbanization level and BMI. Effective measures should be taken to improve the physical quality of college students.