Pathological effects of feeding aflatoxin-contaminated feed on immune status and reproductive performance of juvenile white leghorn males and its mitigation with ∝-tocopherol and Moringa oleifera.

This study was planned to detect the adverse pathological consequences of aflatoxin B1 in White Leghorn (WLH) layer breeder males. Eight-week-old male layer cockerels were separated into six experimental categories: A group was kept as negative control, offered with normal feed only; group B was fed with 400 ppb amount of aflatoxin, while groups F and D fed with normal feed and supplemented with vitamin E 100 ppm and 1% Moringa oleifera, respectively, whereas groups E and C were fed with 400 ppb aflatoxin containing feed and ameliorated with vitamin E 100 ppm and 1% Moringa oleifera, respectively. This study was continued for 2 months and immunologic disorders and reproductive parameters were observed during the trial. To find out immunological status lymphoproliferative response to phytohemagglutinin-P (PHA-P), antibody titers against sheep red blood cells (SRBCs) and carbon clear assay were performed by collecting samples from five birds from each group. The whole data was measured by ANOVA test, and group means were compared by DMR test by using M-Stat C software. Regarding the reproductive status, spermatogenesis, blood testosterone level, testes weight, testes histology, sperm motility, and morphology were negatively affected by aflatoxins, but these deviations positively ameliorated by vitamin E and Moringa. Vitamin E and Moringa found advantageous in boosting the immune status of affected bird. All the immunological parameters including antibody titers against sheed red blood cells, lymphoproliferative response to avian tuberculin and phagocytic potential of macrophages were suppressed by AFB1 however in control, Moringa and vitamin E groups these immunological responses were significantly higher.

Recalling the reported toxicity assessment of deoxynivalenol, mitigating strategies and its toxicity mechanisms: Comprehensive review.

Mycotoxins frequently contaminate a variety of food items, posing significant concerns for both food safety and public health. The adverse consequences linked to poisoning from these substances encompass symptoms such as vomiting, loss of appetite, diarrhea, the potential for cancer development, impairments to the immune system, disruptions in neuroendocrine function, genetic damage, and, in severe cases, fatality. The deoxynivalenol (DON) raises significant concerns for both food safety and human health, particularly due to its potential harm to vital organs in the body. It is one of the most prevalent fungal contaminants found in edible items used by humans and animals globally. The presence of harmful mycotoxins, including DON, in food has caused widespread worry. Altered versions of DON have arisen as possible risks to the environment and well-being, as they exhibit a greater propensity to revert back to the original mycotoxins. This can result in the buildup of mycotoxins in both animals and humans, underscoring the pressing requirement for additional investigation into the adverse consequences of these modified mycotoxins. Furthermore, due to the lack of sufficient safety data, accurately evaluating the risk posed by modified mycotoxins remains challenging. Our review study delves into conjugated forms of DON, exploring its structure, toxicity, control strategies, and a novel animal model for assessing its toxicity. Various toxicities, such as acute, sub-acute, chronic, and cellular, are proposed as potential mechanisms contributing to the toxicity of conjugated forms of DON. Additionally, the study offers an overview of DON's toxicity mechanisms and discusses its widespread presence worldwide. A thorough exploration of the health risk evaluation associated with conjugated form of DON is also provided in this discussion.

Protective efficacy of a novel multivalent vaccine in the prevention of diarrhea induced by enterotoxigenic Escherichia coli in a murine model

Background@#Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. @*Objectives@#Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. @*Methods@#SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). @*Results@#Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. @*Conclusions@#Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

Evaluation of efficacy of tacrolimus in the one-year treatment of vitiligo / 中华皮肤科杂志

Objective@#To evaluate the efficacy of long-term treatment with tacrolimus ointment in patients with vitiligo.@*Methods@#A total of 156 patients with vitiligo (70 males and 86 females) were enrolled into this study, who were firstly diagnosed with vitiligo and continuously followed up in Department of Dermatology, Yiwu Dermatology Hospital and the Third People′s Hospital of Hangzhou between January 2016 and February 2018. Of the 156 patients, 114 were adults, and 42 were children aged 6 - 18 years. All the patients received 6-month treatment with tacrolimus ointment twice a day. They were followed up once a month, the time to initial repigmentation and vitiligo area severity index (VASI) were recorded, and VASI improvement rate was calculated. After 6-month treatment, the patients achieving marked improvement were divided into conventional treatment group and interval treatment group to be treated with tacrolimus ointment twice a day and once every 3 days respectively for another 6 months, and final efficacy of the 2 protocols was compared. Mauchly′s test of sphericity, randomized block analysis of variance and Bonferroni method were used to analyze differences among pre-treatment VASI scores, and 1- to 6-month post-treatment VASI scores.@*Results@#Among the 156 patients, the pre-treatment, and 1-, 2-, 3-, 4-, 5- and 6-month post-treatment VASI scores[M (P25, P75) ] were 2 (0.6, 5) , 2 (2, 5) , 1.6 (0.575, 4.5) , 1.2 (0.5, 4.0) , 0.4 (1.15, 3.5) , 0.75 (0.3, 2.575) , 0.6 (0.2, 2.2) respectively. Additionally, there was a significant difference in the VASI score among the above time points (F = 6.14, P < 0.05) , and the VASI score showed a gradual decreasing trend over time. During the initial 6-month treatment, the average time to initial repigmentation was 2.2 ± 0.6 months. Of the 156 patients, 98 (62.8%) received marked improvement, and the time to marked improvement was 5.2 ± 0.6 months. Of the 98 patients who received marked improvement, 46 completed another 6-month treatment. After 12-month treatment, there was no significant difference in the repigmentation rate between the conventional treatment group (73.3% ± 18.2%) and interval treatment group (72.0% ± 16.2%, t = 0.42, P > 0.05) .@*Conclusions@#The initial 6-month topical tacrolimus therapy showed marked efficacy for vitiligo. After the marked improvement of skin lesions, good efficacy can be maintained by long-term treatment with topical tacrolimus once every 3 days or twice a day.

Evaluation of efficacy of tacrolimus in the one-year treatment of vitiligo / 中华皮肤科杂志

Objective To evaluate the efficacy of long-term treatment with tacrolimus ointment in patients with vitiligo.Methods A total of 156 patients with vitiligo (70 males and 86 females) were enrolled into this study,who were firstly diagnosed with vitiligo and continuously followed up in Department of Dermatology,Yiwu Dermatology Hospital and the Third People's Hospital of Hangzhou between January 2016 and February 2018.Of the 156 patients,114 were adults,and 42 were children aged 6-18 years.All the patients received 6-month treatment with tacrolimus ointment twice a day.They were followed up once a month,the time to initial repigmentation and vitiligo area severity index (VASI) were recorded,and VASI improvement rate was calculated.After 6-month treatment,the patients achieving marked improvement were divided into conventional treatment group and interval treatment group to be treated with tacrolimus ointment twice a day and once every 3 days respectively for another 6 months,and final efficacy of the 2 protocols was compared.Mauchly's test of sphericity,randomized block analysis of variance and Bonferroni method were used to analyze differences among pre-treatment VASI scores,and 1-to 6-month post-treatment VASI scores.Results Among the 156 patients,the pre-treatment,and 1-,2-,3-,4-,5-and 6-month post-treatment VASI scores [M(P25,P75)] were 2 (0.6,5),2 (2,5),1.6 (0.575,4.5),1.2 (0.5,4.0),0.4 (1.15,3.5),0.75 (0.3,2.575),0.6 (0.2,2.2) respectively.Additionally,there was a significant difference in the VASI score among the above time points (F =6.14,P ＜ 0.05),and the VASI score showed a gradual decreasing trend over time.During the initial 6-month treatment,the average time to initial repigmentation was 2.2 ± 0.6 months.Of the 156 patients,98 (62.8％) received marked improvement,and the time to marked improvement was 5.2 ± 0.6 months.Of the 98 patients who received marked improvement,46 completed another 6-month treatment.After 12-month treatment,there was no significant difference in the repigmentation rate between the conventional treatment group (73.3％ ± 18.2％) and interval treatment group (72.0％ ± 16.2 ％,t =0.42,P ＞ 0.05).Conclusions The initial 6-month topical tacrolimus therapy showed marked efficacy for vitiligo.After the marked improvement of skin lesions,good efficacy can be maintained by long-term treatment with topical tacrolimus once every 3 days or twice a day.

Effects of tacrolimus in treating vitiligo on the secretion of local chemokine / 中华医学美学美容杂志

Objective To analyze the abnormalities of local chemokines in patients with vitiligo and to explore the effect of tacrolimus on the secretion of chemokines in keratinocytes.Methods Blister fluids of 50 patients with vitiligo were collected,including lesion areas and normal areas.Luminex was used to analyze the concentration of local chemokines in patients with vitiligo to determine whether the chemokines were closely related to vitiligo.The effect of tacrolimus on chemokine secretion of was analyzed by Western blot in HaCaT cells.Results By Luminex analysis of blister fluid,it was found that CXCL9 and CXCL10 were significantly higher in the leukoplakia of vitiligo,and there was a significant difference,compared with the blister fluid in the normal site (P＜0.01).IFN-γ significantly stimulated the keratinocyte cell line HaCat to express CXCL9 and CXCL10.After pretreatment of HaCaT cells with 20 mg tacrolimus,the expression of CXCL9 and CXCL10 was significantly decreased,compared with the blank control (P＜0.01).Conclusions The leukoplakia chemokines CXCL9 and CXCL10 are highly expressed in vitiligo patients.The tacrolimus can significantly reduce the expression of CXCL9 and CXCL10 in keratinocytes under stress,and it therefore plays a therapeutic role in vitiligo.

Expression of ubiquitin-conjugating enzyme E2S in malignant melanoma and its effect on the biological behavior of melanoma cells / 中华皮肤科杂志

Objective To determine the expression of ubiquitin-conjugating enzyme E2S (UBE2S) in malignant melanoma (MM),and to evaluate its effect on the biological behavior of melanoma cells.Methods Immunohistochemical study was performed to determine the UBE2S expression in 128 primary MM tissue chips,64 metastatic MM tissue chips,16 non-tumor tissue chips (8 paralesional normal skin tissues and 8 normal epidermal tissues).Real-time quantitative RCR was conducted to determine the UBE2S mRNA expression in the melanoma cell lines A375,MUM-2B and MUM-2C.The melanoma cell lines A375 and MUM-2B were divided into 2 groups separately:interference group transfected with a lentiviral vector carrying UBE2S RNA interference sequence,and control group transfected with a lentiviral vector carrying control sequence.After 72 hours,real-time quantitative RCR was performed to determine the UBE2S mRNA expression in the melanoma cell lines A375 and MUM-2B.Caspase-3/7 activity in the groups was assessed by using kits,and cell apoptosis and cell cycle distribution were detected by flow cytometry.The effect of UBE2S knockdown on the migratory and invasive abilities of and N-cadherin expression in A375 cells were evaluated by Transwell assay and Western blot analysis respectively.Statistical analysis was carried out with SPSS 22.0 software by using independent sample t-test for the comparison of normally distributed data between two groups,chi-square test for enumeration data,MannWhitney U test for the comparison of non-normally distributed data,and Spearman's coefficient for assessment of the correlation of UBE2S expression with T staging of melanoma.Results UBE2S was highly expressed in 98 (51.0％) MM tissues,but lowly expressed in 16 non-tumor tissues,and the UBE2Sexpression rate significantly differed between the above two kinds of tissues (x2 =11.905,P ＜ 0.01).UBE2S expression was negatively correlated with T staging of melanoma (ρ =-0.210,P =0.043).The relative mRNA expression of UBE2S significantly differed among the A375,MUM-2B,and MUM-2C cells (F =817.228,P ＜ 0.01).After UBE2S knockdown,the caspase-3/7 activity was significantly up-regulated in the A375 interference group (t =17.572,P ＜ 0.01) and MUM-2B interference group (t =24.552,P ＜0.01) compared with the A375 and MUM-2B control groups respectively.Compared with the control group,the A375 interference group showed significantly increased proportion of A375 cells at G1 phase (t =7.365,P ＜ 0.01),decreased proportion at S phase (t =-9.190,P ＜ 0.01),and no change in the proportion of A375 cells at G2/M phase (t =-0.227,P ＞ 0.05).The MUM-2B interference group showed significantly increased proportions of MUM-2B cells at G1 (t =12.676,P ＜ 0.01) and G2/M phases (t =13.045,P ＜0.01),but significantly decreased proportion at S phase (t =-15.718,P ＜ 0.01) compared with the control group.Transwell assay revealed decreased migratory and invasive abilities of A375 cells in the interference group compared with the control group (t =-35.727,-125.000,P ＜ 0.05,＜ 0.01,respectively).Western blot analysis showed down-regulated expression of N-cadherin protein in A375 cells in the interference group compared with the control group.Conclusions UBE2S is over-expressed in melanoma tissues,whose expression is associated with the T staging of melanoma.Knockdown of UBE2S affects the apoptosis,cell cycle,migration and invasion of melanoma cells,and may promote the metastasis of MM cells by regulating N-cadherin expression.

Clinical analysis of oxytocin combined with misoprostol in preventing postpartum hemorrhage of cesarean section / 中国生化药物杂志

Objective To investigate the clinical effect of oxytocin combined with misoprostol in preventing postpartum hemorrhage of cesarean section.Methods 86 patients with cesarean section who were treated and diagnosed in our hospital from March 2016 to October 2016 were randomly divided into experimental group and control group according to the time of hospitalization.The patients were randomly divided into experimental group and control group.43 cases.Preoperative two groups of patients with different treatment methods for science introduction, so that pregnant women mental stress to relax, ease the heart anxiety and nervous mood.On the basis of conventional treatment, the experimental group received the uterine injection of oxytocin, while the patients under the tongue to take drugs;the control group of patients with oxytocin injection for preventive treatment.Results In the two groups, 32 cases were effective in the experimental group, 1 case was ineffective, the effective rate was 97.67%, compared with 10 cases in the control group, 16 cases were ineffective, the total effective rate was 62.79 % was statistically significant(P<0.05).Conclusion Oxytocin combined with misoprostol has a good therapeutic effect on the postpartum hemorrhage in the prevention and treatment of uterine bleeding.Compared with the original conventional treatment, there is a high efficiency, and the patient's comfort is Higher to improve, to a certain extent, is the more excellent clinical treatment.

Correlation between aortic elasticity and coronary artery calcification evaluated by CT / 中国医学影像技术

Objective To discuss the correlation between aortic elasticity and coronary artery calcification by CT.Methods Totally 111 patients who were diagnosed of coronary artery disease underwent coronary artery CTA.The images were qualified for aortic elasticity measurement.All patients were divided into calcification negative group (n=43) and calcification positive group (n =68).The calcification positive group was further divided into light,medium,and serious groups according to their calcification scores.The ascending aortic images were reconstructed every 5 ％ R-R intervals.The cross-sectional areas and diameters of aortic in each R-R interval were measured automatically,then diameter variation rate (％ A0),aortic distensibility (A0D),aortic compliance (A0C) and aortic stiffness (A0SI) were calculated to evaluate aortic elasticity.Correlation between aortic elasticity and coronary artery calcification were analyzed.Results ％ A0,A0 D,A0C were lower and A0SI was higher in calcification positive group than those in calcification negative group (all P＜0.05).There was no significant differences in the four reference indexes of aortic elasticity among light,medium,and serious groups in calcification positive group (all P＞0.05).Correlation analysis demonstrated negative correlations between ％ A0,A0 D,A0 C and calcification scores,and a positive correlation between A0SI and calcification scores.Conclusion Aortic elasticity is correlated with coronary artery calcification,and the combination of them will be beneficial for the early diagnosis of coronary heart disease.

Curative efficacy of cyclophosphamide combined with VAD regimen in treatment of multiple myeloma and its effects on coagulation function / 中国生化药物杂志

Objective To study curative efficacy of cyclophosphamide combined with VAD regimen in treatment of multiple myeloma and its effects on coagulation function.Methods 78 patients of multiple myeloma who received therapy from July 2013 to July 2016 in our hospital were selected as research objects, and divided into the control group and the observation group , the control group was treated with VAD regimen, while the observation group was treated with cyclophosphamide combined with VAD regimen.Then coagulation factor and anti-coagulation factor activity, adverse reaction, therapeutic effect after treatment between two groups were compared.Results After treatment, decreased blood coagulation factor, Protein C, protein S and antithrombin were increased between two groups , the difference was not statistically significant.Incidence of adverse reactions in observation group was less than the control group[15.38%(6/39)vs 46.15%(18/39)], the difference was statistically significant(P<0.05);The total effective rate of observation group was statistically higher than that in the control group [ 89.74%( 35/39 ) vs 58.97%( 23/39 ) ] , the difference was statistically significant(P<0.05).Conclusion Cyclophosphamide combined with VAD regimen is well for multiple myeloma, which can effectively improve the patient's condition, improve the life quality of patients, and will not cause a greater impact on blood coagulation function.

Discussion on the county -level public hospitals reform from the point of medical care management / 中国基层医药

County -level public hospitals reform is an important measure of deepening reform of the medical and health system.Medicare payments system reform is therefore an important part of public hospitals reform.By abolishing drugs addition,registration fee,treating fee and setting up medical service fee instead,our hospital carried on a diversified management style in the aspect of medical insurance total prepaid system.The implementation of the reform measures turns out a good result:reasonably keeping a control on the medicare spending and really embodying the transformed of interests patterns.On the premise of ensuring patients interests,medical personnel′s interests are improved to a new level and medical insurance fund are also brought under a reasonable control.The medicare payments system reform puts equity in access and people benefit as the starting point and footing of county -level public hospitals reform,thus lays a foundation of further development of public hospitals reform.

Clinical analysis of 41 cases of the shoulder dystocia / 中国基层医药

Objective To explore the high risk factors,clinical prediction and treatment methods of the shoulder dystocia to reduce the incidence of complications in mother and child.Methods The clinical data of 41patients with shoulder dystocia were retrospectively analyzed,and they were divided into the huge baby group and normal weight baby group.The causes,incidence of complications in mother and child and the treatment were ana-lyzed.Results In the 41 cases of shoulder dystocia,the incidence of complications in mother and child in the macro-somia group was higher than the normal weight baby group(the incidence of postpartum hemorrhage,the incidence of neonatal asphyxia in the huge baby group were higher than the normal weight baby group,χ2 =4.439,5.225,all P 0.05,and the difference between the two groups reach the significant level.Conclusion The shoulder dystocia is a serious maternity emergency,the huge baby is the high factor of the shoulder dystocia,the important treatment to reduce the incidence of complications in mother and child is the correct prevention,prediction,early identification and proper treatment.

Expression characteristic of IFNGR in stellate ganglion of female goats / 中国免疫学杂志

Objective:To detect the existence of IFNGR in the stellate ganglion in female goats .Methods:The stellate ganglia were taken from female goats .The expression of IFNGR was detected by PCR method and immunohistochemical SP staining .Results:The results showed that:IFNGR immunoreactive substances were distributed in neurons , supporting cells and passing fiber , and mainly in the cytomembrane and nuclei of neurons .The relative expression of IFNGR was very significantly higher than that of non-neuronal cells (P<0.01).The sequence of IFNGR1 gene amplified by PCR method contained 376 bp, and IFNGR1 gene of goats exhibited the highest homology with sheep(98%),followed by cattle(97%).Conclusion:The results suggested that the IFNGR in the Stellate Gan-glion of female goats mainly expressed and located in sympathetic postganglionic neurons which were provided with the conditions for the role of IFN-γ, which implied that the Stellate Ganglion may act as the critical point to concert the immune regulation of IFN -γand neu-roregulation of autonomic nerve on cardiovascular system .

Expression of progesterone receptor in the cervicothoracic ganglion of female goat / 解剖学报

Objective In order to explore whether neurons in the cervicothoracic ganglion of female goat were equipped with the condition for the role of progesterone .Methods Immunohistochemical SP method was used to detect the expression of progesteron receptor ( PR) in cervicothoracic ganglion of female goats .Results The result indicated that PR-immunoreactive-products were mainly present in the cell bodies of neurons , and satellite cells , Schwann cells , neurapophysis and nerve fiber also had weaker staining .The cell membrane and cytoplasm of neurons represented brown as strong positive ,but the nucleus of neurons showed the presence of heterogeneity , 85.10% neurons ’ nucleus was strong positive,of which 31.91%nucleolus being clear vacuoles were negative;14.90%nucleus displaying vacuolization had no shaining, of which 7.45%nucleolus was strong positive .There were weak positive and flaxen PR-immunoreactive-products in satellite cells, Schwann cells, neurapophysis and nerve fiber .Image analysis shows that PR of neurons was extremely significant compared with other non-nerve cells ( P <0.01 ) . Conclusion The result proved that the sympathetic postganglionic neurons in cervicothoracic ganglion of female goat were the one of main target cells of progesterone ,suggesting that progesterone may be involving in the neuroregulation of cardiac functional activities through affectinng the activities of neurons in the cervicothoracic ganglion of female goat ,and PR in the cervicothoracic ganglion may act as a network node to coordinate endocrine regulation of progesterone and neuroregulation of autonomic nerve on the cardiac functional activities .

Vascular endothelial growth factor C expression is closely correlated with lymph node recurrence and poor prognosis in patients with early stage cervical cancer.

OBJECTIVES: To detect vascular endothelial growth factor (VEGF)-C mRNA expression in surgically resected tissues of 'pathologic N0' (pN0) cervical cancer; to investigate the relevance of VEGF-C mRNA expression to clinicopathological factors, lymph node recurrence and prognosis in early stage cervical cancer. METHODS: Patients with pN0 cervical cancer who successfully underwent radical hysterectomy with bilateral adnexectomy and bilateral pelvic lymphadenectomy were enrolled sequentially into this retrospective study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to detect VEGF-C mRNA. RESULTS: Seventy-eight patients entered the study. VEGF-C mRNA was detected in 35 (44.87%) patients and was significantly correlated with tumour differentiation. VEGF-C mRNA expression was significantly associated with lymph node recurrence and poor overall survival 5 years after surgery. Multivariate analysis confirmed that VEGF-C mRNA expression was an independent predictor for lymph node recurrence and unfavourable overall survival. CONCLUSIONS: These findings indicate that detection of VEGF-C mRNA has clinical potential as a predictor for identifying patients with pN0 cervical cancer at high risk of lymph node recurrence and poor prognosis.

rAAV-mediated VLDLR gene transfection reduces tau hyperphosphorylation in hippocampus of type 2 diabetic rats / 中国病理生理杂志

AIM: Abnormal hyperphosphorylation of tau plays a critical role in the pathogenesis of Alzheimer's disease(AD), and tau protein was hyperphosphorylated in type 2 diabetes. The present study was designed to explore the phosphorylation level of tau in hippocampus of type 2 diabetes rats which interrupted by very low density lipoprotein receptor(VLDLR)gene transfection. METHODS: Wistar male rats were randomized into 3 groups. The control group(CTL)was fed with normal food. The T2DM group and T2DM mediated VLDLR gene group were on high sugar, high fat and high protein diet for 3 months. The plasma insulin level was measured by RIA method, and the plasma glucose was determined by glucose-oxidase method. Total tau level, the phosphorylation level of tau at individual phosphorylation sites and the level of VLDLR were analyzed by Western blotting. The activity of glycogen synthase kinase 3β, a key component of insulin signal transduction pathway and a known tau kinase, in the hippocampus of rats was determined by using [γ-~(32)P]-ATP and the specific peptide substrate. RESULTS: No significant difference of total tau level in hippocampus between T2DM group and T2DM mediated VLDLR gene group was observed. Tau protein in T2DM group was found to be more hyperphosphorylated at several AD-related phosphorylation sites(Ser214, Thr217, Ser396, Ser422 and Ser199/202)than that in CTL, while the immunoreaction at tau-1 site is weaker than that in CTL. VLDLR gene therapy reduced hyperphosphorylation sites of Thr217, Ser396, Ser422 and Ser199/202 of tau to almost the control level, but did not change the phosphorylation of Ser214 or Ser422 on tau. The expression of Ser214 was also observed by immunohistochemical assay. The phosphorylated tau modestly increased in hippocampus in T2DM group compared to CTL, but VLDLR gene treatment did not change the phosphorylation level. The phosphorylation of GSK-3β was decreased dramatically in the hippocampus in T2DM rats, and this phosphorylation was significantly increased after VLDLR gene treatment. CONCLUSION: These findings suggest that Raav mediated VLDLR gene treatment partially reverses tau hyperphosphorylation at several sites in T2DM rat hippocampus, which may mediate by inhibition of GSK-3β activity.

Effects of low-molecular weight heparin on MMP-2, TIMP-2 expression and invasiveness of cytotrophoblastic cells / 中国病理生理杂志

AIM: To investigate the hypothesis that low-molecular weight heparin (LMWH) regulates in vitro cytotrophoblast invasiveness and production of metalloproteinases-2 (MMP-2), tissue inhibitor of metalloproteinas-2 (TIMP-2). METHODS: Chorionic villi tissue of normal 6-8 weeks pregnancy was obtained. Trophoblastic cells were collected by trypsin-collagenase digestion and Percoll gradient centrifugation. The cytotrophoblastic cells were cultured for 24 h and divided into 4 groups according to the concentrations (1.0×10~2 IU/L, 1.0×10~3 IU/L or 1.0×10~4 IU/L) of LMWH adding into the medium. The contents of MMP-2 and TIMP-2 in cell culture supernatants were measured by the method of ELISA. Cytotrophoblast invasiveness was determined by Transwell chamber assay. RESULTS: With the increasing concentrations of LMWH, the invasion activity of cytotrophoblastic cells and MMP-2 secretion were increased. At concentration of 1.0×10~3IU/L, LMWH greatly enhanced cytotrophoblast invasiveness and the expression of MMP-2 (P<0.05). The levels of TIMP-2 were decreased after intervention with LMWH. At concentration of 1.0×10~3IU/L or 1.0×10~4 IU/L, LMWH induced a significant decrease in TIMP-2 expression. No significant difference between group 1×10~3IU/L and group 1.0×10~4 IU/L was observed (P>0.05). CONCLUSION: LMWH might regulate cytotrophoblast invasiveness in vitro by influencing the expression of MMP-2 and TIMP-2 in cytotrophoblastic cells.

Expression and distribution of VLDLR and its subtypes in various tissues of type 2 diabetic rat / 中国糖尿病杂志

Objective To investigate the distribution and expression of total amount and two subtypes of very low density lipoprotein receptor(VLDLR)in various tissues(heart,cerebrum,kidney,muscle,adipose)of normal SD rats and type 2 diabetic SD rats.Methods The mRNA of VLDLR from various tissues were amplified by semi-quantitative RT-PCR.Results The expression levels of VLDLR were reduced in all collected tissues compared with normal ones,especially in cerebrum,kidney,muscle,adipose(P