The corneal epithelial cells are at the surface of the cornea.Human corneal epithelial cells cultured in vitro contribute to fully understand the biological properties of corneal cells,construct disease models in vitro,and play its clinical significance better.This article reviews the recent advances in human corneal epithelial cells cultured in vitro about extractive fraction,extraction methods,factors of progress,impact identification methods.

Dry eye is a common ocular disease with a high morbidity.Many factors contribute to dry eye with cause pathophysiological changes of the ocular surface.In recent years,with the increasing attention of dry eye and the research progress,people have more in-depth understanding of the dry eye.This article reviews the researches in recent years and new progress in the treatment of dry eye.

Objective To explore the efficacy of hormone replacement therapy (HRT) for rheumatoid arthritis (RA).Methods The literature about HRT in the treatment of rheumatoid arthritis were searched.Eleven papers were subjected to a Meta-analysis and a heterogeneity test was conducted to evaluate the efficacy of HRT.Results HRT reduced the level of erythrocyte sedimentation rate (ESR) in RA patients (P=0.016),the SMD was-0.22(-0.40,-0.04); improved bone mineral density (BMD) in RA patients (P=0.022),the WMD was 2.83(0.41,5.26); decreased clinical parameters for disease activity evaluations of RA patients (P=0.048),the SMD was-0.19 (-0.38,0.00); decreased the level of C-reactive protein (CRP) in RA patients,the SMD was-0.08 (-0.37,0.21),but the difference was not statistically significant (P=0.591).Conclusion The findings from this Meta-analysis indicate that HRT can reduce the ESR level,improve clinical indexes and improve BMD level of RA patients.HRT may suppress disease activity and osteoporosis of RA patients,so it may be used as an auxiliary therapy in the treatment of RA.

AIM: To investigate the pharmacokinetic properties and bioequivalence of nifedipine sustained-release tablets after multiple doses administration in healthy volunteers. METHODS: Twenty two male healthy volunteers were enrolled in a randomized two-way crossover design with multiple doses (20 mg·d-1×7 d) study. Nitrendipine was used as the internal standard and the concentrations of nifedipine in plasma were determined by HPLC-APCI-MS. The pharmacokinetic parameters were calculated and the bioequivalence were compared by DAS (ver 1.0) program. RESULTS: The pharmacokinetic parameters of test and reference preparations were as follows: Cmax (52.5±27.4) and (54.0±31.2) ng·ml-1;Cmin (5.4±4.1) and (6.2±5.9) ng·ml-1;Cav (16.8±9.2) and (19.3±12.4) ng·ml-1;Tmax (3.7±0.9) and (4.1±1.1) h;t1/2 (8.9±4.9) and (8.5±3.1) h;AUC0-τ (403.4±221.0) and (461.9±296.6) μg·h·L-1, AUC0-36h (444.4±256.1) and (503.1±330.9) ng·h·ml-1;AUC0-∞ (482.1±268.9) and (542.3±348.4) ng·h·ml-1;DF (299.8±117.7)% and (279.2±97.5)%, respectively. There were no significant differences (P>0.05) in Tmax, Cmax, Cmin, Cav, DF, AUC0-τ, AUC0-36h, AUC0-∞ and t1/2 between the two preparations. The relative bioavailability of test tablets was (100.6±38.6)%. CONCLUSION:The test and reference preparations were bioequivalence.

OBJECTIVE:To establish a method for simultaneously determination of glycyrrhizic acid and glycyrrhetinic acid in Yaotongning capsule. METHODS:HPLC was performed on the column of Agilent TC-C18 with mobile phase of methanol-0.2 mol/L ammonium acetate(gradient elution) at a flow rate of 1.0 ml/min;detection wavelength was 250 nm and column temperature was 25 ℃ and injection volume was 10 μl. RESULTS:The linear range was 0.007 1-0.178 0 mg/ml(r=0.999 8)for glycyrrhizin acid and 0.354 8-8.720 0 μg/ml of glycyrrhetinic acid(r=0.999 8);RSDs of precision,stability and reproducibility tests were no more than 1.74%,average recoveries were 95.49%-100.62%(RSD=1.98%,n=9)and 96.80%-102.26%(RSD=1.83%,n=9),respec-tively. CONCLUSIONS:The method is simple,reproducible,accurate and reliable,and can be used for the simultaneous determi-nation of glycyrrhizic acid and glycyrrhetinic acid in Yaotongning capsule.

Objective：Our aim was to investigate the proportion of autosomal recessive (AR) inheritance among families with patients with Duchenne muscular dystrophy (DMD) and clinical feature in patients with AR form of DMD. Methods:A total of 193 families was studied， 8 of them with at least one girl with “DMD - like” phenotype and 185 with only boys with this kind of phenotype. Based on the number of families with at least one affected girl and the number of patients per sibship among these pedigrees, the proportion of families with DMD inherited as an AR trait was estimated. The clinical examination, family history and serum creatine-kinase were studied in 11 patients diagnosed as AR form of DMD. Results: The proportion of families with AR form of DMD was estimated as 9.4%. The average age of being able to walk is (1.47±1.00) year, serum creatine-kinase levels were (2785.10±1500.29) U/L. The clinical symptom occurred at the average age of (8.11±4.32) year in patients with AR form of DMD. Conclusion： The AR form of muscular dystrophy and DMD not be distingushed clinically. Some families with only affected boys diagnosed as typical DMD, in fact, have the AR form of the disease. This study is very useful for genetic consulting.

Objective To observe the effect of general anesthesia induction assisted dexme-detomidine on blood pressure responses to ephedrine.Methods Forty-four patients scheduled for lap-aroscopic cholecystectomy were randomly divided into normal saline(group N)and dexmedetomidine (group D)group.Group D was treated 15 minutes by micro pump injecting the dose of 0.8 μg/kg dexmedetomidine before anesthesia induction.Then the rate was changed to 0.4 μg·kg-1 ·h-1 and maintained.Meanwhile patients were given anesthesia induction and trachea intubation.0.1 mg/kg ephedrine was injected 5 minutes after trachea intubation.Likewise group N was treated 15 minutes by micro pump injecting physiological saline before anesthesia induction.The other treatments were same.SBP,DBP and HR were recorded before micro pump injecting dexmedetomidine or physiologi-cal saline(T0 ),before anesthesia induction(T1 ),during trachea intubation(T2 ),2 min after trachea intubation(T3 ),during ephedrine injection(T4 ),2 min,5 min,10 min and 15 min after ephedrine (T5 ,T6 ,T7 ,T8 ).Results Compared with T0 ,SBP and DBP of group N was lower at T1 ,T3-T8 but SBP,DBP and HR was higher at T2 (P<0.05 or P<0.01).HR of group N was lower at T4 ,T7 and T8 (P<0.05 or P<0.01).SBP at T1-T8 ,DBP at T1-T4 and T8 ,HR at T1 and T3 ,T4 was lower in group D(P<0.05 or P<0.01).Compared with T2 ,SBP,DBP and HR of group N was lower at T3 and T4 (P<0.01).SBP of group D was lower at T4 (P<0.01).Compared with T4 ,SBP of group N was only higher at T5 and T6 (P<0.05 or P<0.01).SBP,DBP and HR of group D were higher at T5-T7 and SBP was kept higher until T8 (P <0.01).Compared with group N,HR of group D was lower at T1-T3 (P<0.05 or P<0.01),SBP,DBP was lower at T2 (P <0.01)and was kept higher from T5 to T8 (P<0.05 or P<0.01).Conclusion Intubation stress response will be relieved during anesthesia induction with dexmedetomidine,which can amplified ephedrine effect.

Objective To investigate the effect of 17β-estradiol on ketamine-induced long-term cognitive deficits in neonatal rats.Methods 80 SD male rats aged 7 days were randomly divided into group C,V,E,K and K+E,and 16 per group.Group C was intraperitoneally injected with same volume of saline for three consecutive days,Group V was subcutaneously injected with same volume of sesame oil for three consecutive days,Group E was subcutaneously injected with 600 μg · kg-1 17β-estradiol for three consecutive days,group K was intraperitoneally injected with 75 mg · kg-1 ketamine for three consecutive days,group K+E was intraperitoneally injected with 75 mg · kg-1 ketamine in combination with 600 μg · kg-1 17β-estradiol injected subcutaneously for three consecutive days.At 2 months of age,learning and memory abilities were tested with the Mortis water maze.After Morris water naze test,ten rats from each group were decapitated and the prefrontal cortex (PFC) was isolated to detect acetylcholine esterase(AchE) activity with ELISA assay and to measure acetylcholine(Ach) level by hydroxylammonium chloride method.Results The escape latency ((40.26±2.36) s,(30.25±2.20) s,(21.55±2.42) s) and path length((1019.35±58.13) cm,(811.16±27.58) cm,(598.34±34.74) cm) of group K were more than those of group C on the third,fourth aud fifth training days (all P＜0.05),while escape latency ((29.46±2.20) s,(24.86± 2.14) s,(17.20±1.91) s) and path length((913.90±41.89) cm,(729.42±31.36) cm,(487.64±18.61)cm) of group K+E were significantly lower than those of group K(all P＜0.05).On test day 6,rats were subjected to a probe trial,ratio of time spent in the target quadrant ((24.5±2.7) ％) and the number of crossings over previous platform locations (1.9±0.5) in group K were fewer than those of group C (all P＜0.05),while ratio of time spent iu the target quadrant((42.3±3.0) ％) and the number of crossings over previous platform locations(3.5±0.5) of group K+E were more than those of group K (all P＜0.05).The AchE activity((0.69±0.04) U · mg pro-1) in rats PFC of group K was significantly higher than that of group C ((0.52±0.06) U · mg pro-1) (P＜0.05).The AchE activity of group K +E ((0.58±0.12)U · mg pro-1) was lower than that of group K(P＜0.05).The Ach level ((2.59±0.34)mg · g-1) in rats PFC of group K was significantly lower than that of group C ((4.35±0.56) mg · g-1) (P＜0.05).The Ach level of group K+E((3.88±0.61) mg · g-1) was higher than that of group K(P＜0.05).Conclusions These results indicate that ketamine impairs learning and memory abilities as rat matures,while 17β-estradiol treatment improves these impairments by inhibiting AchE activity and increasing Ach level.

Objective To investigate the copy number changes on chromosome 3q26. 1 in urothelial carcinoma of the bladder, and to explore its potential clinical significance. Methods The microarray-based comparative genomic hybridization (Array-CGH) approach was used to analyze the genome-wide copy number changes of 35 tumor tissue samples of bladder cancer. To confirm the loss of a small fragment in 3q26. 1 detected by Array-CGH, real-time fluorescent quantitative polymerase chain reaction (real-time PCR) was performed with 57 frozen tumor tissue samples and 34 formalinfixed paraffin-embedded (FFPE) tumor tissue samples. The urine sediment cells collected from 15 healthy volunteers and 29 bladder cancer patients were checked as above. Results The Array-CGH data showed that the copy number loss of a small fragment in 3q26. 1 was detected in 77.1% (27/35)of the tumor tissue samples investigated. Real-time PCR analysis validated this loss of a small fragment of 3q26.1 with high frequencies in both 57 frozen tumor samples and 34 FFPE tumor samples.The percentage of samples exhibiting loss was 78.9% (45/57) and 100. 0% (34/34) respectively.Furthermore, the relative copy number of the 3q26.1 small fragment was significantly lower in the urinary sediment cells of the patients (median=0. 0020), comparing with that of healthy controls (median=0. 0030) (P＜0.01). Conclusions Loss of the small fragment in 3q26.1 could be a characteristic genetic change of urothelial carcinoma of the bladder. It may serve as a potential molecular marker for bladder cancer.

To screen and identify the possible pathogen of the firstly confirmed human case of avian influenza A in Beijing, the throat swabs and tracheal aspirates of this case were collected and the H5N1 viral nucleotide was tested with real time RT-PCR. The certification of result, screening of other pathogens in respiratory tract and sub-typing of influenza viruses were made by using re-sequencing microarray. It was found that the H5N1 viral nucleic acid was positive in the tracheal aspirate of this case by means of detection with real time RT-PCR and the specific sequence of the non-structural protein (NS) gene of H5N1 virus was obtained through the detection with re-sequencing clip. Through the comparative study with the sequence in Genbank, it was proved to be the H5N1 nucleic acid of avian influenza viruses and excluded the possibility of infections with 30 subtypes of influenza viruses and 33 other respiratory tract pathogens. It is apparent that the pathogen detection with re-sequencing clip shows the high sensitivity and specificity and it plays an important role in the pathogen screening and identification for the firstly confirmed human case of avian influenza A in Beijing.