Objective To determine the changes of caspase 3 and 8 genes and protein expressions during indomethacin (IND) induced gastric mucosal cell apoptosis in vivo . Methods Healthy male SD rats were treated intragastrically with four different doses of IND. The rats were killed 3 hours after IND administration and the TUNEL technique was applied to detect mucosal cell apoptosis. The change of caspase 3 mRNA expression was detected by in situ hybridization and RT PCR techniques and the changes of caspase 3, 8 protein expression were monitored immunohistochemically. Results In the rats of control group, TUNEL assay revealed that only a few apoptotic cells. Oral administration of IND resulted in the appearance of massive TUNEL positive cells. Computer aided image analysis showed that the mean pixels in 30~120mg/kg groups were 6 3 , 8 0 , 12 6 and 17 1 fold that in control group ( P

The aim of the study was to demonstrate the influence of immersion and restraint stress on the ultrastructural changes in gastric acid barrier and parietal cells in rats. Twenty rats were randomly divided into control and stress groups. The gastric mucosal ulcer index was measured. The ultrastructural changes in parietal cells, epithelial cells, epithelial cell junctions, and basal lamina were observed by transmission electronic microscopy. Stress could induce gastric mucosal damage obviously. Parietal cells in a resting state in control group but became active in stress groups with plenty of mitochromosomes and secreting cysts. The cell membrane of epithelium on the luminal surface were were injured with the preservation of tight junction and basal lamina. The results indicated that the stress induced acid secretion of parietal cells, which destroys the luminal surface of the epithelium. It thus suggests the significance of epithelial cell membrane damage in the development of stress ulcer.

SD rats were exposed to single or repeated water immersion restraint stress (WRS) for 4 h every other day for up to 6 days, during which the extent of gastric mucosal lesions was evaluated grossly and histologically. The expressions of EGF and EGFR in gastric mucosa were assayed by immunohistochemistry. The results showed that single exposure to WRS led to haemorrhagic lesions, but tolerance developed following repeated exposures to WRS. Histological evaluations showed that single exposure to stress conditions led to crater like haemorrhagic necrosis almost extending to the muscularis mucosae. The submucosa was apparently congestive and edematous. Following repeated WRS, however, mucosal haemorrhagic necrosis was significantly attenuated and submucosal edema and congestion apparently lessened. Active regeneration and proliferation of mucosal cells were observed at the same time. The expressions of EGF and EGFR could be detected in normal gastric mucosa. EGF was expressed in cytoplasma mainly in the regenerative zone with weaker expression at the basal portions of the gastric glands. EGFR was mainly distributed on the membrane of cells in the regenerative zone. Single WRS led to significant decreased in the expressions of EGF and EGFR. their expressions were absent in the necrotic region. After repeated WRS, enhanced expression of EGF and its receptors could be observed not only in mucosal cells of regenterative zone but also in other areas including the lumen of gastric glands. The results suggested that single WRS could cause severe gastric mucosal lesions, but gastric mucosa became tolerant to repeated WRS by way of toleration cytoprotection. Gastric adaptation was accompanied by active regeneration of mucosal cells mediated by EGF binding with its receptor, EGFR.

BACKGROUND: Retinitis pigmentosa (RP) is a non-inflammatory, bilaterally progressive, retinal degeneration characterized by loss of photoreceptor cells via an apoptotic mechanism, and it eventually leads to blindness.Research shows that the traditional Chinese medicines of Astragalus has great prospect on blocking the progression of RP disease.OBJECTIVE: To observe the protective effect of Astragalus on N-methylN-nitrosourea (MNU)-induced retinal damage in Sprague-Dawley (SD) rats and provide the optimal treatment for RP in humans.DESIGN: Randomized controlled experiment.SETTING: School of Pharmaceutical Sciences, Xinxiang Medical College.MATERIALS :The experiment was completed in Pharmacological Laboratory of Zhongshan Ophthalmic Centre, Sun Yat-sen University between March to December 2004. Totally 114 female SD rats were purchased from the Animal Center of Zhongshan Medical College, Sun Yat-sen University.MNU was purchased from Sigma Company of America. Astragalus injection was purchased from Chengdu Diao Jiuhong Pharmaceutical Factory (Batch No. Z99060535, 2 mL/ampoule, main ingredient: Astragalus).METHODS: Among 114rats, 30 were for morphometric analysis of retinal layers, 30 were for detection of apoptosis and 54 were for detection of NF-κB p65 activity. All of them were randomly divided into different groups and each group had 6 rats. Astragalus at doses of 2.5, 5 and 10 g/kg were injected intraperitoneally into 47-day rats once a day. Meanwhile, a single intraperitoneal injection of 60 mg/kg MNU was given to 50-day rats in model and Astragalus groups. At different intervals after MNU treatment,the animals were sacrificed. Retinal damage was evaluated based on retinal thickness, the apoptotic index of the photoreceptor cells was detected by TUNEL labeling and the DNA-binding activity of NF-κB p65 was analyzed according to transcription factor assay kit.MAIN OUTCOME MEASURES: Comparison of retinal thickness, apoptotic index and the activity of nuclear NF-κB p65.RESULTS: Totally 114 rats entered the result analysis. Pretreatment with Astragalus could dose-dependently suppress MNU-induced photoreceptor cell loss and decreased the apoptotic index. Astragalus at dose of 10 g/kg also time-dependently up-regulated the activity of nuclear NF-κB p65.However, protective effect of Astragalus on MNU-induced central retinal damage was not found.CONCLUSION: Astragalus partially protects against MNU-induced retinal damage by up-modulating the activity of nuclear NF-κB p65 to inhibit apoptosis of photoreceptor cells in a dose-dependent manner.

BACKGROUND: Hemiplegia caused by stroke or brain injury may lead to secondary osteoporosis. Under the circumstance, bone mineral density (BMD) and Bone Mineral Content (BMC) are two effective factors for predicting the risk of osteoporosis-related fracture OBJECTIVE: To investigate the characteristics of secondary osteoporosis in stroke patients with hemiplegia by measuring and analyzing BMD and BMC.DESIGN: A case study.PARTICIPANTS: From March 1999 to May 2002, 156 hemiplegia patients at the Neurorehabilitation Department of Peking Boai Hospital were selected, 87 males and 69 females, with an age ranging from 17 to 65 years old, in average of (47.0±5.3) years old and having a course of diseases ranging (5.0±3.5) months. The types of their illness: 89 had cerebral infarction, 67 had cerebral hemorrhage. Hemiplegia sides: 79 patients had hemiplegia in the left and 77 in the right.METHODS: Using LUNAR Dual Energy X-Ray densitometers, all the patients underwent a fro-top-to-toe scan to assess the BMD and BMC status.MAIN OUTCOME MEASURES: ① The occurrence rate of osteoporosis in patients with various course of hemiplegia. ② The BMC in patients with hemiplegia in various part.RESULTS: All the 156 patients entered the statistical analysis procedure.① The occurrence rate of osteoporosis was highest in patients with 3 to 6 months of hemiplegia (44%, 16/36) and was lowest in those with 9 to 12 months of disease (21%,7/33). The total occurrence rate of osteoporosis was 31% (48/156). ② For those with a less-than-3-month course of hemiplegia: Of all the 47 patients, 9 had dystonic hemiplegia, all suffering from osteoporosis and 2 of whom having fracture. For those with a 3-to-6-month course of hemiplegia: Of all the 36 patients, 3 had dystonic hemiplegia and osteporosis as well. ③ BMC in the affect side of upper limb was signifi cantly lower than that in healthy side [(154.76±43.91) g, (172.59±43.78) g,t=3.591, P＜ 0.001]. BMC in the affect side of lower extremity was close to that in healthy side [(463.41±30928) g, (464.11±86.45) g, t=0.027, P＞ 0.05].CONCLUSION: In patients with hemiplegia, changes in bone mass fluctuate with the length of hemiplegia. BMC in the affect side of upper limb is low while that in the healthy side is nearly normal. It may be explained that upper extremities start the functional exercises at an early time. This indicated that bone mass has a close relationship with excercises.

Objective To explore how Vibrio vulnificus (Vv) invades dendritic cells (DC) and induces acute necrosis of DC via toll-like receptor (TLR) 2 and 4 pathways.Methods Vv 1.1758 strain and DC 2.4 mixed culture model was established,observed the infection rates of DC with optical microscope,the location of Vv and structural changes of DC by transmission electron microscope.The expression levels of TLR2 and TLR4 mRNA were determined by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and tumor necrosis factor-α (TNFα) protein titers were measured by enzyme-linked immunosorbent assay (ELISA).DNA ladder qualitative test was used to detect cell apoptosis,while flow cytometry was used to quantify cell apoptosis and necrosis rates.Statistical analysis was done by chi-square test and one-way ANOVE.Results The infection rates of DC after 0.5,1,2,4 and 6 h of mixed culture were (7.8±0.8) ％,(13.9± 1.1) ％,(34.6±4.9) ％,(77.8± 10.2)％ and (95.8 ± 13.1)％,respectively.Vv was generally located in the internal cell membrane of DC 2.4.After 2 h co-culture,nuclear chromatins of DC became active and intranuclear apoptosis bodies appeared.After 4 h,cytoplasmic vacuoles appeared,chromatin gathered,and cell membranes were seriously damaged.After 6 h,mitochondria was highly swelled and distorted,and cell apoptosis and necrosis occurred.TLR2 and TLR4 mRNA levels reached peak values after co-culture for 0.5 h; TNF-α level began to increase at 1 h (P＜0.05) and reached peak values at 2 h.DNA Ladder electrophoresis presented scouring necrosis after 2 h culture and apoptotic bands appeared between 720 bp and 900 bp after 4 to 5 h culture.Early apoptosis rates of DC after 2,4 and 6 h culture were (3.1±3.8)％,(7.8±4.7)％ and (12.7±8.2)％,and necrosis rates of DC were (16.7±12.5)％,(41.6±25.9)％ and (75.5±33.6)％,higher than that of control group (all P＜0.05).Conclusions Vv infects DC and induce DNA degradation through up-regulated expression of TLR2 and TLR4 and increasing of TNF-α inflammatory mediators.During cell degradation,apoptosis and necrosis coexist,while necrosis is predominant.

Objective To investigate the variance levels of plasma soluble leukocyte differentiation antigens CD40 (sCD40) and soluble CD40 ligand (sCD40L) in preeclamptic patients with renal damage and its relationship. Methods A total of 63 pregnant women attended the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College between August 2008 and June 2010. In the present study included 28 pregnant women with mild preeclampsia and 35 patients with severe preeclampsia. Thirty matched normotensive pregnant women were enrolled in the study as the control group. Expression of sCD40 and sCD40L were determined by ELISA. At the same time, the blood routine, C reaction protein ( CRP),urine routine, 24 hours urine protein excretion, and serum uric acid (UA), creatinine (Cr), blood urea nitrogen (BUN) were measured. The correlation analysis was performed between the sCD40/sCD40L and the blood biochemical indexes in 3 groups. Results ( 1 ) The median levels of CRP in severe preeclampsia (10. 8 mg/L)and mild preeclampsia group(7. I mg/L)are significantly higher than that of control group (3. 3 mg/L,P ＜ 0. 05 ); The level of CRP in severe preeclampsia group was also higher than that of mild preeclampsia group ( P ＜ 0. 05 ). The median gestational age at delivery in severe preeclampsia ( 32. 5 weeks)was significantly less than that of mild preeclampsia group ( 37. 2 weeks) and normal group ( 38. 6 weeks,P ＜ 0. 05). However no significant differences were observed between mild preeclampsia group and normal group ( P ＞0. 05 ). The platelet count in severe preeclampsia ( 132 × 109/L) was significantly less than those of mild preeclampsia group (212 × 109/L) and normal group ( 216 × 109/L, P ＜ 0. 01 ), but no significant differences were observed in blood platelet amount between mild preeclampsia group and normal group ( P ＞0. 05 ). There was no significant difference in hemoglobin level and white blood cell in three groups ( P ＞0. 05). (2) The sCD40 plasma concentration in severe, mild preeclampsia and normal group was 133.6,126. 5 and 90. 7 ng/L, respectively. The sCD40 L plasma concentrations were 12. 5, 10. 4 and 4. 4 ng/L respectively in the 3 groups. 24 hours urinary protein quantitative was 4. 5 g/d,0. 8 g/d and 0 in the 3 groups respectively. And the UA level was 486 μ mol/L,289 μmol/L and 162 μmol/L. In the above three groups,the monitoring indicators were significantly higher in women with severe preeclampsia group compared with mild preeclampsia and control groups (P ＜ 0. 01 ), and there were also higher in mild preeclampsia group than that in control groups ( P ＜ 0. 01 ). The level of plasma Cr ( 89 μmol/L) and BUN ( 5. 32 mmol/L) in severe preeclampsia group were higher than those of mild preeclampsia group (66 μmol/L and 4. 49mmol/L) and control group ( 57 μmol/L and 3.32 mmol/L, P ＜ 0. 05 ). There was no significant difference between mild preeclampsia group and normal group (P ＞ 0. 05 ). (3) The correlation analysis indicated that the level of sCD40 has a positive correlation with 24 hours urinary protein quantitative( r = 0. 434, P ＜ 0. 05 ),also significant positive correlation( r =0. 536,0. 528 ,P ＜ 0. 01 ) between the level of sCD40 and UA or CRP in women with preeclampsia. There was no significant correlation between the level of sCD40 and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count(r =0. 135,0. 183, -0. 133,0. 190,0. 167, -0. 221 ,all P ＞0. 05 ). There were positive correlation between the level of sCD40L and 24 hours urine protein excretion, either UA or CRP( r =0. 591,0. 445,0. 539 ,all P ＜0. 01 ). No significant correlation was found between sCD40 L and systolic blood pressure, diastolic blood pressure,delivery gestational age, Cr, BUN, and platelet count( r =0. 178,0. 212, -0. 292,0. 144,0. 135, -0. 273,all P ＞0. 05). There was significant positive correlation between plasma sCD40 and sCD40L ( r =0. 707 ,P ＜0. 01 ). There was no relationship between the level of sCD40, sCD40L and the blood biochemical indexes in normotensive pregnant women ( P ＞ 0. 05 ). Conclusions The plasma concentrations of sCD40 and sCD40 L are significantly higher in pregnant women with preeclampsia compared with the control, which may be involved in the development of preeclampsia and contribute to the kidney damage. The variance levels of sCD40 and sCD40L may be also related to the severity of preeclampsia.

Adult ; Altitude Sickness ; metabolism ; pathology ; Apoptosis ; BH3 Interacting Domain Death Agonist Protein ; metabolism ; Erythroblasts ; cytology ; pathology ; Humans ; Male ; Middle Aged ; RNA, Messenger ; genetics ; bcl-2-Associated X Protein ; metabolism ; bcl-X Protein ; metabolism

Adult ; Aged ; Altitude ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Point Mutation ; Polycythemia ; genetics

Mucin as an important member in gastrointestinal mucus layer has become the focus of research. Many studies suggested that MUC1 mucin is involved in the mechanism of Helicobacter pylori( Hp)infection. It is important to understand the relationship between MUC1 mucin and Hp for preventing Hp infection and gastric cancer. This article reviewed the relationship between MUC1 mucin and Hp.