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.

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

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.

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

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.

To explore the new method of discriminating Cistanche deserticola, Cynomorium songaricum and Orobanche pycnostachya by using PCR amplification of specific alleles. 30 samples of the different C. deserticola, 21 samples of C. songaricum and O. pycnostachya were collected. The total DNA of the samples were extracted, the ITS sequences from C. deserticola, C. songaricum and O. pycnostachya were amplified by PCR and sequenced unidirectionally. These sequences were aligned by using ClustulW. Specific primer was designed according to the ITS sequences of specific alleles, and PCR reaction system was optimized. Additionally, compare with the identification of specific PCR method and DNA sequence analysis method. The result showed that the 331 bp identification band for C. deserticola and the adulterants not amplified bands by a single PCR reaction, which showed good identification ability to the three species. PCR amplification of specific alleles can be used to identify C. deserticola, C. songaricum and O. pycnostachya successfully.
Alleles ; Cistanche ; classification ; genetics ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Phylogeny ; Polymerase Chain Reaction ; methods

In this paper, Liuwei Dihuang pill was used to study the identification of Chinese patent medicine by fluorescence sequencing typing technology. The DNA of Paeonia suffruticosa was used as template to amplify by five pair of FAM fluorescence labeling primers. Then, the amplified products were sequenced. The sequencing results were analyzed by GeneMarker V1.80 to screen the best fluorescence labeling primers. As a result, psbA-trnH fluorescence labeling primer was used to identify the raw materials of Liuwei Dihuang pill. The results showed that three kinds of raw plant medicinal materials in Liuwei Dihuang pill were able to be correctly identified by psbA-trnH fluorescence labeling primer. The fluorescence sequencing typing technology can stably and accurately distinguish raw medicinal materials in Chinese patent medicine.
DNA Primers ; chemistry ; genetics ; DNA, Plant ; chemistry ; genetics ; Drugs, Chinese Herbal ; chemistry ; standards ; Fluorescent Dyes ; chemistry ; Plants, Medicinal ; chemistry ; genetics ; Polymerase Chain Reaction ; instrumentation ; methods ; Quality Control ; Staining and Labeling

The sequences of ITS, matK, rbcL and psbA-trnH of 9 Gynostemma species or variety including 38 samples were compared and analyzed by molecular phylogeny method. Hemsleya macrosperma was designated as outgroup. The MP and NJ phylogenetic tree of Gynostemma was built based on ITS sequence, the results of PAUP phylogenetic analysis showed the following results: (1) The eight individuals of G. pentaphyllum var. pentaphyllum were not supported as monophyletic in the strict consensus trees and NJ trees. (2) It is suspected whether G. longipes and G. laxum should be classified as the independent species. (3)The classification of subgenus units of Gynostemma plants is supported.
Gynostemma ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Sequence Analysis, DNA