Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

ObjectiveTo study the quality control methods for 99Tcm-TRODAT-1 kit and injection.MethodsThe appearance,pH,contents of the bases,the labeling yield,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 kits from three different batches were examined.The kit stability was estimated under different conditions.The transparence,pH,radiochemical purity,half-life,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 injection were tested.ResultsThe 99Tcm-TRODAT-1 kit and injection were both achromous and transparent,with pH values being 5.9 ± 0.1 and 5.5 - 7.0 respectively.The contents of stannous chloride and TRODAT-1 were stable.The labeling yield of the kit and the radiochemical purity of the injection were both ≥95％.The asepsis test demonstrated that the characters of 99Tcm-TRODAT-1 kit and injection were qualified.TRODAT-1 kit was stable at 0 -4 ℃ for 6 months or at room temperature (20 -25 ℃ )for 10 days,and the radiochemical purity of the injection was still ＞ 90％ at room temperature for 8 hours.ConclusionsThe quality control methods for 99Tcm-TRODAT-1 kit are simple and practical.The kit and injection are qualified and can be used for clinical application.

Background The incidence of retinal vascular diseases increase annually,such as diabetic retinopathy,retinopathy of prematurity and age-related macular degeneration.The key of treatment for these diseases is how to evaluate retinal vascular change effectively and objectively.Retro-orbital injection of fluorescein isothiocyanatedextran (FITC-dextran) is a simple and effective method for observing C57BL/6J mouse retinal vessels.But,whether it is suitable for other mice and rats is seldom reported.Objective This experiment was to assess the feasibility of the observation of retinal vessels by retro-orbital injection of FITC-dextran in different genus of mouse and offer the reference for relevant study.Methods Twelve animals of C57BL/6J mice,Kunming mice,SD rats and Wistar rats were selected,respectively and divided into the experimental group and control group at average.The right eyes of the animals of the experimental group received the retro-orbital injection of 9 ml/kg FITC-dextran,and the right eyes of animals of the control group received PBS solution at the same volume and way.All the animals were sacrificed 10 seconds after injection and both eyes of each animal were obtained for retinal stretched preparation.The retrobulbar tissue and whole-mount retina were viewed under a fluorescence microscope.The use of the animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Retinal blood vessels labeled by FITC-dextran could be observed in both eyes of C57BL/6J mice and Kunming mice to present with a green fluorescence in experimental group under a fluorescence microscope,but no any fluorescence-labeled retinal blood vessel was exhibited in the control mice.The retinal blood vessel could not be observed in all eyes of SD rats and Wistar rats after the injection of FITC-dextran both in the experimental group and the control group under a fluorescence microscope.The surrounding tissues of the right eyes of mice and rats dyed with green fluorescence of FITC-dextran in the experimental group,however,green fluorescence could not be seen in the surrounding tissues of the left eyes of mice and rats.Conclusions Retro-orbital injection of FITC-dextran is a suitable method of observing the retinal vessels of mouse but not rat.

OBJECTIVETo study the effects of Tongxinluo ultramicro-pulverization (TXLU) on experimental myocardial infarction and platelet aggregation of rats, investigate its mechanisms on ischemia heart disease and offer a reference to clinical usage.

METHODRats were separated randomly into 7 groups: sham, model, diltiazem (0.15 mg x kg(-1)), TXL(1.2 g x kg(-1)), TXLU (1.2, 0.6, 0.3 g x kg(-1)). The experimental myocardial infarction was induced with ligating the left anterior descending branch of the coronary of rats. The infarction size was determined after myocardium tissue was stained with 2,3,5-triphenyltetrazolium chloride (TTC). And the serum of rats was separated to analyze CK, LDH, SOD, MDA. Another 60 rats were separated randomly into 6 groups: control, aspirin (0.15 mg x kg(-1)), TXL (1.2 g x kg(-1)), TXLU (1.2 ,0.6,0.3 g x kg(-1)). The rat platelet aggregation was induced with adenosine diphosphate (ADP) and collagen to observe the inhibitory effects of TXLU.

RESULTTXLU could relieve the myocardial infarction size and weight stained with TTC significantly, the myocardial infarction size of the three groups of TXLU were (2.7 +/- 2.1)%, (3.4 +/- 1.2)%, (2.8 +/- 1.8)%, compared with model group (8.9 +/- 5.9)%, P < 0.05 or P < 0.01. The myocardial infarction weight of the three groups of TXLU were (8.4 +/- 3.5)%, (8.7 +/- 4.1)%, (9.7 +/- 4.1)%, compared with model group (l2.2 +/- 3.6)% P < 0.05 or P < 0. 01. And the content of MDA and the activities of CK and LDH in rats subjected with ligation of coronary artery were inhibited obviously too, compared with model group P < 0.05 or P < 0.01, then the activity of SOD increased. TXLU could inhibit the maximum percentage of rats platelet aggregation induced with ADP and collagen, the maximum percentage of platelet aggregation induced with ADP were (26.9 +/- 9.2)%, (24.4 +/- 13.4)%, (30.6 +/- 12.2)%, compared with control group (44.3 +/- 15. 7)% P < 0.05 or P < 0.01; The maximum percentage of platelet aggregation induced with collagen were (33.8 +/- 6.9)%, (32.1 +/- 8.3)%, (41.5 +/- 7.8)%, compared with control group (49.2 +/- 15.9)%, P < 0.05 or P < 0.01.

CONCLUSIONThe experiment results indicated that TXLU could protect myocardial tissue of rats from ischemic injury and the mechanism may be related with antioxidation and inhibiting platelet aggregation, and the results also suggested TXLU could lower clinical dosage.

Adenosine Diphosphate ; pharmacology ; Animals ; Aspirin ; pharmacology ; Diltiazem ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Medicine, Chinese Traditional ; methods ; Myocardial Infarction ; blood ; chemically induced ; drug therapy ; pathology ; Platelet Aggregation ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; pharmacology

OBJECTIVETo investigate the status of voice fatigue and throat disease in the secondary school teachers in Urumqi.

METHODSThis study included 11 689 teachers from 109 secondary schools offered by Teaching and Researching Center of Urumqi as the second-stage random sampling. Among them, 3217 teachers were selected to carry out voice fatigue test and routine examination of throat. For those with abnormal routine inspection results, the stroboscopic laryngoscopy were added.

RESULTSThe failure rate difference of 3217 secondary school teachers voice fatigue tests during different periods had statistical significance (F = 202.653, P < 0.05). The failure rate difference of voice fatigue test among different throat diseases groups had statistical significance (F = 9.516, P < 0.05). There was statistical significance in failure rates of the voice fatigue tests between 908 cases of disease group and 2309 cases of normal group among different genders and periods (P < 0.05). Female teachers had higher failure rates than male teachers in voice fatigue tests. High failure rates appeared when high volume required for male and female teachers in voice fatigue tests. There was statistical significance in mean value sound pressure level (MVSPL) and mean value fundamental frequency (MVF0) of the voice fatigue tests between different gender and periods (P < 0.05). Male teachers had higher MVSPL than female teachers, female teachers had higher MVF0 than male teachers. There was statistical significance in failure rates of the voice fatigue tests among teachers from different tribes in different periods (P < 0.05). The Uighur had the highest failure rate, while Han had the lowest. Make further pairwise comparisons, failure rates difference of the Han and the Uighur in the voice fatigue tests during different periods had statistical significance. Comparison among other tribes had no statistical significance (P > 0.05).

CONCLUSIONSVoice fatigue is common in secondary school teachers. Female teachers tend to get voice fatigue more easily, when required high volume, the voice fatigue is more serious. Uighurs teachers have highest degree of voice fatigue.

Adult ; Faculty ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; diagnosis ; epidemiology ; Teaching ; Voice Disorders ; diagnosis ; epidemiology ; Young Adult

Objective To explore the features of connected speech produced by Chinese mild Alzheimer's disease (AD) patients.Methods Thirty Chinese mild AD patients (eight males and 22 females,and the age was (72.73 ± 7.78) years) and 30 age and education matched normal controls from three communities were recruited in Xuanwu Hospital from January 2018 to May 2018.All subjects completed tasks of picture naming,semantic fluency (animal),and Cookie Theft picture description.Picture description was analyzed for speech production (total word output,speech rate,sound errors),lexical content (number of nouns and verbs,proportion of pronouns and verbs,semantic errors),information content (information unit,idea density,efficiency),syntactic structure and complexity (mean length of utterance,words in sentences,syntactic errors).Differences between groups were calculated.Association between picture naming,semantic fluency,and measures in picture descriptions was analyzed.Results Compared with the controls,mild AD patients achieved lower scores on the picture naming task (74.45±8.04 vs 79.60± 1.73,t=-2.800,P=0.010) as well as on the semantic fluency task (10.35±3.08 vs 18.50±4.66,t=-6.520,P＜0.01) and produced lower speech rate (131.80±36.40 vs 149.64±26.69,t=-2.095,P=0.041),less number of nouns (7.52±2.55 vs 9.67±3.58,t=-2.509,P=-0.015) and verbs (4.40±2.24 vs 5.73±1.92,t=-2.375,P=0.021),increased proportion of pronouns (0.286 (0.319) vs 0.039 (0.122),Z=3.602,P＜0.01) and semantic errors (0.629 (1.396) vs 0 (0.779),Z=2.386,P=0.017),less information units (11.00±3.63 vs 13.70±3.53,t=-2.787,P=0.007),decreased idea density (0.11±0.04 vs 0.16±0.05,t=-3.946,P＜0.01),decreased efficiency (14.23±7.56 vs 22.95±7.14,t=-4.738,P＜0.01),shorter mean length of utterance (6.64± 1.27 vs 8.25± 1.70,t=-3.928,P＜0.01),and less words in sentence (0.84±0.10 vs 0.95±0.06,t=-4.625,P＜0.01).Group differences in total word output,sound errors,proportion of verbs,and syntactic errors did not reach statistical significance.Scores on picture naming task were negatively associated with the proportion of pronouns (r=-0.737,P=0.003),and positively associated with the idea density (r=0.540,P=0.046) and efficiency (r=0.592,P=0.026) in the AD group.Scores on semantic fluency task were negatively associated with the proportion of pronouns (r=-0.554,P=0.011).Conclusions Connected speech in mild AD patients was characterized by decreased naming accuracy,decreased sematic fluency,decreased content words,information units,conciseness,efficiency,and syntactic complexity,but motoric aspects of speech and sentence structure remained relatively preserved.Decreased content words and increased pronouns in mild AD may relate to semantic impairment.

OBJECTIVE: To explore the effect of electroacupuncture on heroin seeking behavior and FosB expression in relevant brain regions. METHODS: Rat model of heroin relapse behaviors was developed with progressive fixed ratio program,and model rats were randomly divided into 3 groups: a restraint group, a needle retention group, and a electroacupuncture group. The heroin seeking behavior was elicited by a small dose of heroin. FosB expression in relevnt brain region was assessed with immunohistochemical technique. RESULTS: Tests on reinstatement of drug seeking behavior induced by heroin priming showed that compared with the restraint group, active pokes in the electroacupuncture group decreased significantly(P<0.05). Compared with the restraint group, the expression of FosB positive nuclei in Acd, Pcg and CeA of rats brain both in the electroacupuncture group and the needle retention group (P<0.05) decreased significantly. In LC, the expression of FosB positive nuclei in the needle retention group decreased significantly compared with the restraint group (P<0.05). CONCLUSION Continuous acupuncture and needle retention attentuate the reinstatement of heroin-seeking behaviors induced by heroin priming, and the inhibitory effect may be mediated partially by the expression of FosB in relevant regions which are involved in the process of heroin addiction.
Amygdala ; metabolism ; Animals ; Behavior, Animal ; Brain ; metabolism ; Electroacupuncture ; methods ; Heroin Dependence ; metabolism ; psychology ; therapy ; Male ; Nucleus Accumbens ; metabolism ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the prevalence and distributing feature of chronic heart failure (CHF) in adult population of Xinjiang.

METHODSFour-stage random sampling method was used to analyze the prevalence and distributing feature of self-reported congestive heart failure among different nationalities in adult (35 years and over) population of Xinjiang. Sampling was collected from 6 localities (Urumqi, Kelamayi, Fukang, Turfan Basin, Hetian, Yili Hazakh).

RESULTSA total of 8459 adults were surveyed. The prevalence of CHF was 1.26% in this cohort. The prevalence of CHF was 0.89%, 1.11% and 2.14%(*) in Han, Uygur and Hazakh population, respectively ((*)P < 0.05 vs. Han and Uygur). The risk of CHF was higher in the males than in the females (1.61% vs. 0.93%, u = 2.79, P < 0.05). The prevalence of CHF increased in proportion with aging and was 0.29%, 0.60%, 1.32%, 2.55% and 4.10% in 35 - 44, 45 - 54, 55 - 64, 65 - 74, 75 years and over age groups, respectively. Common complications of CHF were hypertension (63.55%), coronary heart disease (42.99%), diabetes (18.69%), valvular heart disease (5.61%) and atrial fibrillation (4.67%).

CONCLUSIONThe prevalence of CHF in Xinjiang was higher than the average level in China (0.9%) and was the highest in the Hazakh population.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; China ; epidemiology ; Chronic Disease ; Female ; Heart Failure ; epidemiology ; Humans ; Male ; Middle Aged ; Prevalence

Objective To investigate the relationship between stroke and ankle-brachial index (ABI), pulse wave velocity (PWV) in Han, Uighur, and Kazakh populations of Xinjiang. Methods Data was from the investigation of cardiovascular risk survey (CRS) program among different nationalities in Xinjiang from October 2007 to March 2010. A total of 14 618 samples aged over 35 with complete data dimension were surveyed. Results ( 1 ) There were 633 patients with stroke, including 258 Hans (4.48%), 247 Uighurs (5.18%), and 128 Kazakhs (3.13%). The incidence of stroke was significant different in the three ethnic groups (P＜0.001). (2)Compared to the Han population, the incidence of stroke was higher ( OR= 1.304) in Uighur but lower (OR=0.794) in Kazakh. (3) PWV was significant different between stroke and non-stroke patients. After adjustment for age, body mass index, systolic blood pressure, triglyceride, total cholesterol and other risk factors, the difference remained significant, indicating that PWV (OR=1.001, P＜0.001) might associate with the occurrence of stroke. Conclusion The results of this study showed that the incidence of stroke was significantly different in the three ethnic groups. PWV might associate with the occurrence of stroke.