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OBJECTIVE@#To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD).@*METHODS@#Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice.@*RESULTS@#The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group (P<0.05). Serum DLA was significantly lower in the YD group than HCD-P group (P<0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group (P<0.01). There was no statistical significance in the serum level of DAO among groups (P>0.05).@*CONCLUSIONS@#YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.
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Mice , Male , Animals , Lactic Acid/pharmacology , Intestinal Mucosa , Colon/pathology , Dexamethasone/pharmacology , Diet, High-Protein , Pneumonia/pathologyРеферат
Six dimensional syndrome differentiation theory, put forward by professor GU Xiao-hong at Beijing University of Chinese Medicine based on her theoretical teaching and clinical experience, emphasizes that the syndrome differentiation should be carried out from six dimensions including etiology, disease location, disease stage, disease condition, pathology, and disease nature, which is conducive to clinical thinking training and formation of traditional Chinese medicine (TCM). The differentiation and treatment of Baihutang syndrome frequently seen in cold damage and warm disease still need to be explored. Guided by the six dimensional syndrome differentiation theory coupled with diverse viewpoints of cold damage and warm disease schools, this paper summarized and reinterpreted the understandings and thoughts of GU Xiao-hong and YU He, warm disease specialists of two generations. Considering the lung-stomach dysfunction caused by the internal invasion of exogenous pathogens, Baihutang syndrome was staged into Qi aspect. In this stage, exuberant pathogens and sufficient healthy Qi allowed the prevailing of internal heat and the consumption of body fluid, manifested as high fever, profuse sweating, thirst, and the pulse corresponding to interior excess and heat syndrome. This paper also pointed out that the Baihutang syndrome involved both lung and stomach, and the adoption of Baihutang contributed to preventing tu from restricting shui in the case of extreme excess of Yang brightness and protecting the kidney Yin. As revealed by the dynamic analysis of prognosis of Baihutang syndrome based on the six dimensional syndrome differentiation theory, even though the Baihutang syndrome could be present in both cold damage and warm disease, the specific disease stage, transmission and change, condition, prognosis, pathology, and medication differed. On this basis, a series of prescriptions have been modified from Baihutang, which has expanded the application scope of Baihutang and enriched its research value, thus better promoting its clinical application.
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·AIM: To investigate the role of small interference RNA interference targeted Integrin-linked kinase (ILK SiRNA) on the proliferation and apoptosis of human Tenon fibroblasts (HTFs) induced by transforming growth factor-β2(TGF-β2). · METHODS: The HTFs were identified by immunofluorescence analysis with Vimentin and keratin. HTFs with no other addiction was as normal control;H+T group:HTFs+5μ g/L TGF-β2;H+T+NC SiRNA group:HTFs+5μ g/L TGF-β2+50nmol/L negtive SiRNA; H+T+ILK SiRNA group:HTFs+5μ g/L TGF-β2+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μ g/L TGF - β2. The protein expression of ILK were analyzed by Western Blot. The proliferation levels of HTFs were analyzed by CCK-8, the apoptosis of HTFs were analyzed by Hoechst 33342/PI double staining. ·RESULTS: The protein ILK were expressed in both TGF-β2treated and control groups, and TGF- β2up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(P< 0. 05). CCK- 8 analysis showed that compared with the normal control group,the cell proliferation rate of HTFs in TGF-β2treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h (P<0.05). Hoechst 33342/PI double staining showed that there was no change on the apoptosis of TGF - β2 stimulated group (P>0.05), compared with the normal control group, however in the ILK SiRNA group, we found lots of apoptosis cells and a few of necrotic cells (P<0.05). ·CONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF - β2, thereby enhancing the apoptosis of HTFs.
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Objective To investigate whether paraptosis and autophagy have an effect on acute retinal ischemia-reperfusion injury (RIRI) in an experimental rat model that recapitulates features of acute hypertensive glaucoma and to explore the possible underlying mechanisms.Methods A total of 30 adult male Sprague-Dawley rats were randomly divided into RIRI group and control group.The acute RIRI model was induced with normal saline in the right eye of rats from the RIRI group by anterior chamber perfusion,while the rats in the control group left untreated.On day 1,day 3,day 7,day 28 after RIRI model establishment,the changes in morphology of retinal ganglion cells (RGCs) were observed by transmission electron microscopy (TEM),and the expression of microtubule-associated protein 1 light chain 3 (LC3) was measured by immumofluorescence methods.Results When compared with the control group,the number of cytoplasmic vacuoles predominantly derived from the progressive swelling of mitochondria and/or endoplasmic reticulum (ER) in RGCs were increased in the RIRI group from day 1 to day 28 by TEM.And ultra-structural analyses showed the double-or multiple-membrane autophagosomes were markedly accumulated in the cytoplasm of RGCs following acute RIRI.The average number of autophagic vacuoles in the cytoplasm of RGCs was 0.79 per 50 μm2 in the control group,and the average number of autophagosomes reached to a maximum on day 7 after acute RIRI at 2.29 per 50 μm2,which was statistically significant compared with the control group (P < 0.05).Compared to the control group,LC3 expression in the cytoplasm of RGCs was up-regulated on day 1 after acute RIRI,which sustained throughout the experimental period.The percentage of LC3 positive cells in the retinal ganglion cell layer was 15.90% in the control group,and the data was 46.95% and 52.30% on day 1 and day 28 after RIRI,respectively,both which were statistically significant compared with the normal control group (both P < 0.05).Conclusion Both paraptosis and autophagy participate in death of RGCs after acute RIRI.Programmed cell death in different cells,either coexistence of multiple-cell death form or a single-cell death form,participates in the pathogenesis of acute RIRI.
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AIM:To compare the effect of three types of pterygium surgery and on tear film in patients with type 2 diabetes mellitus. ·METHODS:A total of 102 patients ( 102 eyes ) with pterygium combined with type 2 diabetes mellitus treated in our hospital from March 2013 to March 2016 were analyzed retrospectively. The patients were divided into three groups including the 34 cases ( 34 eyes ) with simple excision of pterygium ( resection group ) , pterygium excision combined with conjunctival flap transplantation in 34 cases (34 eyes, as conjunctival flap group ) and pterygium excision combined with limbal stem cell transplantation in 34 cases ( 34 eyes, as stem cell group ) . The wound repair time, complications, recurrence rate, uncorrected visual acuity (UCVA), tear film break-up time ( BUT ) and basal tear secretion test (SⅠt) were observed before, and 6 and 12mo after surgery in the three groups, respectively. ·RESULTS:The postoperative UCVA of the three groups was significantly higher than that preoperation ( P =0. 039, 0. 013, 0. 024 ), and there was no significant difference among the three groups ( P = 0. 317 ). The wound repair time was 5. 67 ± 1. 45d in the resection group, which was significantly higher than that in the conjunctival flap group (4. 18 ± 0. 76d) and the stem cell group (4. 09±0. 79 d) (P<0. 001), there was no significant difference between the conjunctival flap group and the stem cell group ( P = 0. 937 ). There were 4 cases in resection group reappeared, and the recurrence rate was 11. 8%, which was significantly higher than the other two groups ( P = 0. 037 ). There were 1 recurrences in the conjunctival flap group, and the recurrence rate was 2. 9%, while the patients in the stem cell group had no obvious recurrence. SⅠt and BUT increased significantly after operation (P<0. 05), especially in conjunctival flap group and stem cell group (P<0. 001). There was no significant difference between the conjunctival flap group and the stem cell group (P=0. 845, 0. 894). · CONCLUSION: Pterygium excision combined with conjunctival flap transplantation or limbal stem cell transplantation for the treatment of type 2 diabetic patients with normal blood glucose and tear film function has the similar effect, and is better than simple pterygium excision.
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@#AIM: To discuss the effect of the cystotome-assisted prechop technique performed in hard nuclear cataract phacoemulsification. <p>METHODS: One hundred and twenty-six patients(158 eyes)of age-related cataracts with nucleus density at grade Ⅲ-Ⅳ were randomly divided into two groups. Group A was performed a manual prechop technique using a surgeon-bent cystotome after the capsulorhexis, while group B was performed traditional phaco-chop without prechop technique. The average power(AP), actual ultrasonic time(U/S time), accumulated energy complex parameter(AECP)of machine, average density of endothelial cells, endothelial cells loss, uncorrected visual acuity(UCVA), corneal edema and intraoperative complications were compared between groups. <p>RESULTS: The AP, U/S time and AECP of Group A were significantly lower than that of Group B(<i>P</i><0.05). At postoperative day 1, the corneal edema of Group A was slighter than the control with significant difference(<i>P</i><0.05), so was the UCVA. While there was no significant difference of UCVA between groups at postoperative 1wk. The average corneal endothelium density of Group B was significantly lower than that of Group A at postoperative 1wk, and the average cell loss was higher than that of Group A. Two eyes of group A had posterior capsular rupture compared to 4 eyes of Group B.<p>CONCLUSION: Compared with traditional phaco-chop, the cystotome-assisted prechop technique presents shorter intraoperative ultrasound time and lower energy, while contributes to less corneal endothelial cell loss and better early postoperative UCVA.
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<p><b>BACKGROUND</b>Retinal degenerative diseases are the leading causes of blindness in developed world. Retinal progenitor cells (RPCs) play a key role in retina restoration. Triamcinolone acetonide (TA) is widely used for the treatment of retinal degenerative diseases. In this study, we investigated the role of TA on RPCs in hypoxia condition.</p><p><b>METHODS</b>RPCs were primary cultured and identified by immunofluorescence staining. Cells were cultured under normoxia, hypoxia 6 h, and hypoxia 6 h with TA treatment conditions. For the TA treatment groups, after being cultured under hypoxia condition for 6 h, RPCs were treated with different concentrations of TA for 48-72 h. Cell viability was measured by cell counting kit-8 (CCK-8) assay. Cell cycle was detected by flow cytometry. Western blotting was employed to examine the expression of cyclin D1, Akt, p-Akt, nuclear factor (NF)-κB p65, and caspase-3.</p><p><b>RESULTS</b>CCK-8 assays indicated that the viability of RPCs treated with 0.01 mg/ml TA in hypoxia group was improved after 48 h, comparing with control group (P < 0.05). After 72 h, the cell viability was enhanced in both 0.01 mg/ml and 0.02 mg/ml TA groups compared with control group (all P < 0.05). Flow cytometry revealed that there were more cells in S-phase in hypoxia 6 h group than in normoxia control group (P < 0.05). RPCs in S and G2/M phases decreased in groups given TA, comparing with other groups (all P < 0.05). There was no significant difference in the total Akt protein expression among different groups, whereas upregulation of p-Akt and NF-κB p65 protein expression and downregulation of caspase-3 and cyclin D1 protein expression were observed in 0.01 mg/ml TA group, comparing with hypoxia 6 h group and control group (all P < 0.05).</p><p><b>CONCLUSION</b>Low-dose TA has anti-apoptosis effect on RPCs while it has no stimulatory effect on cell proliferation.</p>