Background Retinal flatmount of oxygen-induced retinopathy (OIR) animal models is a useful tool in the study of ischemic retinopathy.The retinas of OIR of rat or mouse pups were small and thick and difficult in operating of conventional preparation and quantitative analysis of retinal flatmounts.Objective This study was to explore an easy and stable operating method of retinal flatmount combined with immunofluorescence staining in rodent.Methods Forty ＜6-hour-old SD rat pups were randomly assigned to OIR model group and normal control group.The pups were raised with nursing mothers in hyperoxia environment (80％) and normal oxygen environment (21％) alternately at a 24-hour interval for 14 days in the OIR model group,and the pups were raised in the room air for 14 days in the normal control group.The eyeballs of the rats were extracted to isolate the retinas intactly.The retinas were stained with glutamine synthetase (GS)-isolectin B4 firstly and then expanded into flatmounted and cut into 4 petals radially.Adobe Photoshop CS3 imaging analysis system was used to match the pictures into entire retinal vascular images and analyzed under the fluorescence microscope.The pixel values of retinal avascular areas and the entire retina were quantified by this system.The percentage of avascular areas to the entire retina was calculated to analyze the severity of non-perfusion areas.The use and care of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results An intact and smooth retinal flatmount could be obtained by firstly staining method.Ora serrata structure was seen surrounding the whole retina.Strong green fluorescence was exhibited in retinal vessel net with clearly visible vascular branches;while the background fluorescence was weaker.Fully developed blood vessels were displayed in the retinas of the normal control group.Non-capillary areas around the central optic disk and large peripheral avascular areas could be seen in the retinal flatmounts of OIR models.Conclusions The preparation of retinal flatmount is easy and feasible by first immunofluorescence staining for retinal vessels followed by radially cutting of retina.This method of retinal flatmount can ensure the integrity of retinal vascular system and it is available for the observation and evaluation of retinal vascular structure in OIR models.

Clinical biochemical test course construction in the new medical model, first of all, requires a combination of subject characteristics of the course, and proceeds with the improvement from the teaching methods and evaluation system. Secondly, we should cultivate students' doctor-patient communication skills and humanities quality in the teaching process. Finally, we should establish the effective clinical biochemical test teaching mode to achieve both teaching and learning.

Objective To evaluate the effect of Sengstaken-Blakemore tube in assisting cyanoacrylate injection combined with sclerotherapy and variceal ligation under emergency endoscopy in treating escophageal and gastric variceal bleeding. Methods 60 patients were randomly selected, and were evenly divided into 2 groups single blindly. Endoscopic cyanoacrylate injection, sclerotherapy and variceal ligation were performed in control group, and additional Sengstaken-Blakemore tube were used in experimental group. Results All the 30 patients in study group got effective hemostasis (30 of 30, 100.00 %), whereas only 23 patients in control groups stopped bleeding (23 of 30, 76.67 %) (P < 0.05). escophageal and gastric variceal were cured 19 patients in the experimental group (19 of 30, 63.33 %) , and only 4 were cured in the control group (4 of 28, 14.28 %) (P < 0.05). Meanwhile, occurence of fever, chest pain, hospital days and hospitalization costs of experimental group was significantly lower than that in control group (P < 0.05). Conclusions Treating the escophageal and gastric variceal bleeding, using endoscopic cyanoacrylate injection, sclerotherapy; and variceal ligation under emergency endoscopy combined with Sengstaken-Blakemore tube could significantly increase the therapeutic efficiency as well as reduce complications and therapy cost.

With the goal of practice teaching for laboratory medicine students, we focus on culti-vating students' clinical thinking and scientific thinking through setting up reasonable teaching task and designing the scientific graduation thesis, and try to develop a novel practice teaching mode for laboratory medicine students. Therefore, we do exploration and practice in the following aspects: orientation training, clinical practice, medical ethics education, clinical communication ability, and scientific research design, hoping to improve students' research thinking ability and innovation ability. Through actual operation and graduation thesis writing, we try to help students to establish and improve their clinical thinking ability and innovation ability.

Objective To investigate the effects of endotoxin on the activation of nuclear factor-kappaB in myocardium, and to explore the molecular mechanism of acute myocardial injury. Methods Forty male Wistar rats were randomly divided into normal control group (n=10), lipopolysaccharide (LPS) 1h, 2h, 6h groups (n=10 for each group). In the latter two groups, LPS was injected into the peritoneal cavity. Tissue samples from the myocardium were collected to determine NF-?B activation by electrophoretic mobility shift assay (EMSA), and the mRNA expression of TNF-? was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) with GAPDH as internal standard. Results The activation of NF-?B (2.173?0.063) and TNF-? mRNA expression (0.292?0.031) could be detected in myocardium in very low values in normal control group rats, while both the activation and expression were up-regulated markedly after LPS challenge, and peaked 2 hours afterwards (37.793?4.785 and 1.182?0.146, respectively). They were down-regulated (17.910?3.791 and 0.901?0.128, respectively) 6 hours after LPS challenge. Compared to normal controls, both NF-?B activity and TNF-? mRNA expression were significantly elevated at various time points following LPS stimulation (all P

Objective To investigate the changes in Toll-like re ceptor 4 (TLR4) and myeloid differential protein-2 (MD-2) gene expression in t he liver of septic rats, and to elucidate the relationship between the expressio n of TLR4/MD-2 mRNA and TNF? mRNA. Methods Sepsis model was reproduced in rats by intraperitoneal injection of LPS (5mg/kg). 80 Wistar rats were randomly divided into 4 groups: normal control and 1h, 2h, 6h after LPS in jection (20 rats for each group). Rats were sacrificed at different time points, and the expression of TLR4/MD-2 mRNA and TNF-? mRNA in the liver was measure d by RT-PCR. Results The expressions of TLR4/MD-2 mRNA and TN F-? mRNA were up-regulated, peaking 1h after LPS injection, and then declined gradually 2~6h after LPS challenge. The expressions at every time point were hi gher than that in control group (P

Adult ; Bacteremia ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Nontuberculous Mycobacteria ; Postoperative Complications

BACKGROUND: When the dynamic equilibrium between oxidation system and anti-oxidation system in the body is upset, the overly produced active oxygen-derived free radicals will attack the target organs, thus resulting in the damage to organs and big molecules, and then diseases. Total anti-oxidation capacity (T-AOC), superoxide dismutase (SOD), and nitric oxide synthase (NOS) activity are the majorindexes for the defense system of the body, and their status is closely related to diseases. OBJECTIVE:To explore the level of T-AOC, SOD and NOS in serum,and find out its relationship with lifestyle. DESIGN:Single sample investigation. SETTING:Department of Health Laboratory Technology, College of Public Health of Harbin Medical University. PARTICIPANTS:Totally 531 residents of Bin County aged 20 to 70years were recruited between September 2000 and May2001. MATERIALS:The reagents kits of T-AOC, SOD and NOS were manufactured by Nanjing Jiancheng Bio-engineering Institute. METHODS:The investigators surveyed the residents with the same questionnaire. Questionnaire was made and the investigators were trained in advance. The questionnaire was filled in item by item as required. Questionnaire items consisted of the general situation, financial status, dietary habit, hobby, status of health care, and mental factor. Finally 95 investigation indexes were completed. The activity of serum T-AOC, SOD and NOS of the 531 residents in Bin County was determined, respectively, with reagent kits produced by Nanjing Jiancheng Bio-engineering Institute. The lifestyle factors were analyzed by multivariate stepwise regression analysis,and informed consent was obtained from the participants. MAIN OUTCOME MEASURFS:① Serum T-AOC, SOD and NOS in the participants; ② lifestyle factors affecting T-AOC, SOD and NOS.RFSULTS:According to actual analysis, the 531 participants entered the results analysis. T-AOC was measured in 489 participants, SOD in 525participants, and NOS in 531 participants. ① Indexes of the participants:T-AOC of the males was obviously higher than that of the females [(20.01±7.21), (15.25±6.22) kU/L, P ＜ 0.05]. SOD of the males was slightly lower than that of the females while NOS of the males was slightly higher than that of the females, but there was no significant difference. ②Multivariate stepwise regression analysis showed that risk factors related to T-AOC were gender difference (OR=2.188), educational level (OR=1.859),and the presence of rheumatism, respectively (OR =1.142). SOD-related risk factors were educational level (OR=1.584), years of spirit drinking (OR =1.048), presence of nephritis (OR=1.093), and irradiation (OR=1.770);frequency of tea drinking was a beneficial factor (OR=0.800). NOS-related risk factors were the average amount of cigarette smoking (OR=1.194) and the times of weekly spirit drinking (OR=1.368). However, the beneficial factor that affected serum NOS was the frequency of weekly mutton eating (OR=0.458) CONCLUSION:This experiment revealed that the ability to clean free radical in the 531 subjects was good and that it was better in males than in females. The increased educational level and presence of rheumatism can decrease T-AOC. Drinking, smoking and irradiation decrease SOD activity while the frequency of tea drinking and mutton eating are beneficial factors.

Objective To synthesize glycylproline p-nitroanilide tosylate as a substrate for glycylprolyl dipeptidyl-aminopeptidase(GPDA),and to study its clinical application.Methods The title compound was prepared by DCC-HOBt synthesis.GPDA in human serum was detected by continuous monitoring method using substrate. Results The substrate with purity of 98.5% was obtained.The linearity of the method was up to 358.1 U/L.Intraassay CV and interassay CV of GPDA in diverse serum samples were 3.01% and 5.04%,respectively.It was shown that GPDA level in patients with hepatic cancer was significantly increased,while those in patients with gastric carcinoma and chronic gastritis were significantly decreased compared with that in the control group in clinical detecting(P

Objective To investigate the influence of gender differences on NF-kB activation in livers in septic rats. Methods Total 20 female and 20 male Wistar rats were randomly divided into four groups.Tissue samples of the livers were collected to measure NF-kB activation by EMSA.The level of plasma ALT,TNF-?and estrogen were measured also. Results NF-kB activation in normal male and female rats has no significant difference (P＞0.05).After stimulated by LPS,the level of NF-kB activation and ALT,TNF-?in plasma were markedly upregulated,and the index of female group lower than that in male group (P＜0.01).The level of NF-kB activation in livers and ALT,TNF-?in plasma both in male and female have significantly negative correlation with the level of estrogen in plasma (P＜0.05 ).Conclusion There are significantly gender differences on NF-kB activation in livers in septic rats.Estrogen may decrease the injury of livers in septic rats.