Research Progress and Forensic Application of Postmortem Genetic Testing in Hereditary Cardiac Diseases / 法医学杂志

Hereditary cardiac disease accounts for a large proportion of sudden cardiac death (SCD) in young adults. Hereditary cardiac disease can be divided into hereditary structural heart disease and channelopathies. Hereditary structural heart disease mainly includes hereditary cardiomyopathy, which results in arhythmia, heart failure and SCD. The autopsy and histopathological examinations of SCD caused by channelopathies lack characteristic morphological manifestations. Therefore, how to determine the cause of death in the process of examination has become one of the urgent problems to be solved in forensic identification. Based on the review of recent domestic and foreign research results on channelopathies and hereditary cardiomyopathy, this paper systematically reviews the pathogenesis and molecular genetics of channelopathies and hereditary cardiomyopathy, and discusses the application of postmortem genetic testing in forensic identification, to provide reference for forensic pathology research and identification of SCD.

Skin Ultrastructure and the Changes of HIF-2α, H-FABP Expression in the Myocardium of Electric Shock Death Rats / 法医学杂志

Objective To observe the skin ultrastructure change of electric shock death rats and to test the expression changes of hypoxia-inducible factor-2α （HIF-2α） and heart type-fatty acid-binding protein （H-FABP） of myocardial cells, in order to provide basis for forensic identification of electric shock death. Methods The electric shock model of rats was established. The 72 rats were randomly divided into control group, electric shock death group and postmortem electric shock group. Each group was divided into three subgroups, immediate （0 min）, 30 min and 60 min after death. The skin changes of rats were observed by HE staining, the changes of skin ultrastructure were observed by scanning electron microscopy, and the expression of HIF-2α and H-FABP in rats myocardium was tested by immunohistochemical staining. Results The skin in the electric shock death group and postmortem electric shock group had no significant difference through the naked eye or by HE staining. Under the scanning electron microscope, a large number of cellular debris, cells with unclear boundaries, withered cracks, circular or elliptical holes scattered on the cell surface and irregular edges were observed. A large number of spherical foreign body particles were observed. Compared with the control group, the expression of HIF-2α in all electric shock death subgroups increased, reaching the peak immediately after death. In the postmortem electric shock group, HIF-2α expression only increased immediately after death, but was lower than that of electric shock death group （P<0.05）. Compared with the control group, the expression of H-FABP in all subgroups of electric shock death group and postmortem electric shock group significantly decreased. The expression of H-FABP in all subgroups of electric shock death group was lower than that of the postmortem electric shock group （P<0.05）. Conclusion Electric shock can increase HIF-2α expression and decrease H-FABP expression in the myocardium, which may be of forensic significance for the determination of electric shock death and identification of antemortem and postmortem electric shock.

Therapeutic observation on tuina at head and abdomen for headache due to phlegm turbidity / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of tuina at head and abdomen for headache due to phlegm turbidity. Methods:A total of 56 patients with headache due to phlegm turbidity were randomized into a tuina group and a Chinese medicine group by the random number table, with 28 cases in each group. The tuina group was treated mainly with tuina at the head and abdomen, while the Chinese medicine group was treated with oral administration of Ban Xia Bai Zhu Tian Ma Tang(Pinellia, Atractylodes Macrocephala and GastrodiaDecoction). The course of treatment was 30 d. The scores of headache index, traditional Chinese medicine syndrome scale, and the therapeutic efficacy were observed. Results:There were 2 dropouts in each group during treatment. The total effective rate was 92.3％ in the tuina group, significantly higher than 76.9％ in the Chinese medicine group (P<0.05). The scores of headache index and traditional Chinese medicine syndrome scale in both groups decreased after treatment (bothP<0.05), and scores in the tuina group were lower than those in the Chinese medicine group (bothP<0.05). Conclusion:Tuina mainly at head and abdomen is effective in treating headache due to phlegm turbidity, and has a better effect than Ban Xia Bai Zhu Tian Ma Tang (Pinellia, Atractylodes Macrocephala and GastrodiaDecoction).

Characteristics of collateral circulation in adult moyamoya disease based on modified Suzuki staging / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the characteristics of collateral circulation in adult moyamoya disease (MMD).</p><p><b>METHODS</b>The clinical data were collected from all adult patients with MMD undergoing digital subtractive angiography (DSA) in our department from 2006 to 2016. Based on the imaging findings, the patients were divided into ischemia group and bleeding group. A double-blind analysis was conducted of the CT or magnetic resonance imaging findings and the severity of the disease was graded using the modified Suzuki score (mSS). We classified the anastomotic networks in MMD into the superficial meningeal type and deep parenchymal type. The superficial meningeal type was further classified into the leptomeningeal and the durocortical networks, and the deep parenchymal networks into subependymal networks and the inner striatal and inner thalamic networks.</p><p><b>RESULTS</b>No significant difference was found in the distribution of mSS scores between the hemorrhage group and the ischemic group (Χ=5.812, v=5, P=0.325), but the posterior communicating artery and internal carotid artery diameter ratio (Pcom/ICA ratio) was significantly greater in the hemorrhage group (t=2.119, v=108, P=0.036). The Pcom/ICA ratio differed significantly among the groups with different mSS scores (f=8.924, P=0.00), higher in groups with mSS scores of 3, 4 and 5. The incidence of anterior choroidal artery dilation differed significantly between hemorrhage and ischemic groups (Χ=11.79, P=0.001). The incidences of durocortical networks (Χ=0.327, P=0.567) and subependymal networks (Χ=0.011, P=0.917) were comparable between hemorrhage group and ischemic groups, but the incidence of leptomeningeal networks (P=0.018) and inner striatal and inner thalamic networks (Χ=7.551, P=0.006) differed significantly between the two groups.</p><p><b>CONCLUSION</b>The collateral circulation vascular system is an important component of cerebral blood flow in MMD patients and varies from patient to patient. Patients with MMD exhibit increased Pcom/ICA ratio with abnormal expansion of the anterior choroidal artery, and the leptomeningeal networks and the inner striatal and inner thalamic networks are independent risk factors for cerebral hemorrhage.</p>

Effects of Huoxue Huayu Decoction combined with musculus pterygoideus lateralis closure on patients with temporomandibular joint dysfunction syndrome due to wind-cold stasis / 中成药

AIM To investigate the clinical effects of Huoxue Huayu Decoction (Angelicae sinensis Radix,Saposhnikoviae Radix,Schizonepetae Herba,etc.) combined with musculus pterygoideus lateralis closure in managing patients with temporomandibular joint dysfunction syndrome (TMJ) due to wind-cold stasis.METHODS From January 2015 to December 2015,68 cases of patients with TMJ due to wind-cold stasis identified and recruited by the department of stomatology of Hebei Provincial Hospital of Traditional Chinese Medicine were randomly assigned to treatment group and control group,each with 34 cases and given the musculus pterygoideus lateralis closure.But the treatment group was administered with an extra two-week adjuvant therapy of Huoxue Huayu Decoction,and generally one week such medication was taken as a therapeutic course.RESULTS After the intervention,the treatment group presented an overall better performance than the control group in terms of the open limit,NRS score,and the differences were statistically significant (P ＜ 0.05);and the values of CMI,PI,DI (P ＜0.05),in contrast to the not significantly varied values between the groups before the intervention (P ＞ 0.05).The treatment group also displayed its advantage if evaluated by TCM criteria,extent of the mandibular movementabnormalities,facial pain,temporomandibular joint sound or noise,irritability,sleeping difficulty,dark purple lips,tongue scores and wind-cold stasis scores (P ＜ 0.05).And generally a remarkably higher total effective rate was observed in the treatment group,and its difference to the control group was statistically significant (P ＜0.05).CONCLUSION Huoxue Huayu Decoction combined with conventional western medicinal approach in the management of TMJ due to wind-cold stasis can achieve a better therapeutic efficacy.

Estimation on Formation Time of Thrombus / 法医学杂志

OBJECTIVES@#To observe the changes of the formation time of venous thrombus in rats, and to provide new ideas and methods for the estimation on thrombus formation time of the forensic cases died from thrombosis.@*METHODS@#Totally 80 rats were randomly divided into 10 groups （0 h, 3 h, 6 h, 12 h, 1 d, 3 d, 1 week, 2 weeks, 3 weeks and 4 weeks after operation）. A vein thrombosis model was established by the "narrow" method. The processes of thrombosis, organization, recanalization and the features of change on hemosiderin and calcium salt were observed by HE stain, Perls stain and Von Kossa stain. The expression changes of CD61, α-SMA and CD34 were observed by immunohistochemical staining technique.@*RESULTS@#Platelets adhered to the exposed blood vessel intima 3 h after operation, and platelet trabeculae were formed by the repeated accumulation of platelets 1 d after operation. The thrombus organization formed through the fibroblasts from vessel wall that grew into the interior of the thrombus 3 d after operation. Endothelial cells covered the surface of thrombus and then the new blood vessels were reformed, and the vessels were reconstructed. The expression of CD61 upregulated at the stages of the thrombus formation （3 h） and thrombus reformation （4 weeks）, and reached the peak 1 d after thrombus formation. The release of hemosiderin and the initial expression of α-SMA were detected 3 d later. Calcium deposit and expression of CD34 were observed 1 week later.@*CONCLUSIONS@#The hemosiderin, calcium salt, CD61, α-SMA and CD34 show time-dependent changing characteristics, which is expected to provide a reference for the estimation on thrombus formation time of the forensic cases died from thrombosis.

Correlating oxidative stress-related factors with bone metabolic markers in elderly male patients exhibiting degenerative osteoporosis in the high-altitude hypoxic region of China: study protocol for a non-randomized controlled trial / 中国组织工程研究

BACKGROUND: Hypoxia is an important factor that affects bone formation and regulates bone growth. Therefore, many elderly patients living in high-altitude hypoxic areas exhibit osteoporosis. Oxidative stress-related hypoxia-inducible factors can induce abnormal expression of various factors including vascular endothelial growth factor (VEGF),insulin-like growth factor, and endothelin. However, it remains unclear whether these factors influence changes in bone metabolic markers.OBJECTIVE: To investigate the correlation between oxidative stress-related factors and bone metabolic markers in elderly male patients with degenerative osteoporosis who reside in the high-altitude hypoxic area of China.METHODS: This is a prospective, single-center, non-randomized, controlled trial. One hundred and twenty elderly male patients with degenerative osteoporosis residing in the high-altitude area of China who receive treatment at the Affiliated Hospital of Qinghai University of China are included as osteoporosis group; 120 healthy elderly males who concurrently receive physical examination are included as control group. One day after admission, serum levels of hypoxia-inducible factor 1-alpha (HIF-1α), HIF-2α, VEGF, osteocalcin, and tartrate-resistant acid phosphatase 5 b (TRACP 5 b) were measured using an enzyme-linked immunosorbent assay. Bone mineral density in L1-4 segments, right femoral neck, and the greater trochanter of the femur was detected using dual-energy X-ray absorptiometry. The primary outcome measure of this study is serum HIF-1α levels at 1 day after admission. Secondary outcome measures include serum levels of HIF-1α, HIF-2α, VEGF, osteocalcin, and TRACP 5 b at 1 day after admission, as well as the correlation between serum levels of oxidative stress indicators (HIF-1α, HIF-2α, and VEGF) and bone metabolic markers (osteocalcin and TRACP 5 b) at 1 day after admission. This study was approved by the Ethics Committee of the Affiliated Hospital of Qinghai University of China (approval No. QHY1402G). The study is performed in accordance with the Declaration of Helsinki.Participants are informed of the study protocol and procedures, and signed an informed consent. Participant recruitment,blood sampling, and data collection are performed during January 2015 to February 2018. Outcome measure analysis and trial completion will be in March 2018. Results will be disseminated through presentations at scientific meetings and/or by publication in peer-reviewed journals. This trial was registered with the Chinese Clinical Trial Registry (Registration number: ChiCTR-ROC-17012848).DISCUSSION: Findings from this study aim to validate the correlation between oxidative stress-related factors and bone metabolic markers in elderly male patients with degenerative osteoporosis who reside in the high-altitude area of China.We intend to confirm risk factors of degenerative osteoporosis in elderly males living in high-altitude hypoxic areas, thus providing guidance for preventing osteoporosis occurrence and development in high-altitude hypoxic areas of China.

Efficacy and safety of Willis covered stent for treatment of blood blister-like aneurysm / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the safety and short-term efficacy of Willis covered stent for treatment of blood blister-like aneurysms (BBA).</p><p><b>METHODS</b>Eight patients with BBA were treated with Willis covered stent system during the period from December 2014 to February 2016. The guiding catheter was placed as high as possible to facilitate the delivery of the covered stent system.</p><p><b>RESULTS</b>s Nine covered stents were implanted in the aneurysms of 8 patients (8 aneurysms), and 8 stents were released successfully in the parent arteries. In 6 patients, angiography immediately after stent release showed complete disappearance of the aneurysm and the parent arteries remained patent. One patient experienced a minor endoleak after stent implantation, and another stent was implanted to eliminate the endoleak. Iatrogeniccarotid-cavernous fistula occurred in 1 patient due to tortuosity of the parent artery, for which superficial temporal artery-to-middle cerebral artery bypass combined with parent artery occlusion was performed instead; the patient recovered smoothly and the bypass remained patent at 6 months after the operation. No other periprocedural complications occurred in these patients. Follow-up study showed no new-onset neurological deficits in these 8 patients, who had mRS score of 0 in 6 patients and of 1 in 2 patients. Digital subtractive angiography at 6 months after the operation demonstrated no aneurysm in these patients, and only one patient showed mild stenosis in the parent artery.</p><p><b>CONCLUSION</b>Willis covered stents are effective for treatment of BBA with good safety and short-term outcomes.</p>

Onyx embolization for treatment of dural arteriovenous fistula: comparison of long- distance versus routine injection method / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the efficacy, clinical characteristics, safety, injection time and radiation exposure of Onyx embolization using a long-distance injection method and routine injection method for management of dural arteriovenous fistula (DAVF).</p><p><b>METHODS</b>The clinical data were retrospectively analyzed in 59 patients with DAVF treated with Onyx embolization using long-distance injection method (28 patients) and routine injection method (31 patients). The efficacy, safety, injection time and radiation exposure during Onyx embolization were compared between the two injections methods.</p><p><b>RESULTS</b>The average radiation dose exposure to the surgeon per procedure was significantly lower in the long-distance injection group than in the routine group. The injection time (P=0.53), injection volume (P=0.78), number of supply arteries (P=0.80), Cognard types (P=0.67), and effect of embolization (P=0.88) were all similar between the two groups.</p><p><b>CONCLUSION</b>Endovaseular treatment of intracranial DAVF with Onyx embolization using the long-distance injection method is feasible, safe and effective and can reduce the radiation exposure to the surgeon.</p>

Cardiac Ultrastructure and Changes of HSP70 and HIF-1α Expression in Electric Shock Death Rats / 法医学杂志

OBJECTIVE@#To observe cardiac ultrastructure and the expression of heat shock protein 70 (HSP70) and hypoxia inducible factor-lα (HIF-lα) in electric shock death rats and to explore the application of these indexes as the basis of medical identification in electric shock death.@*METHODS@#Seventy-two SD rats were randomly divided into electric shock death group, postmortem electric shock group and the control group. The changes of myocardial ultrastructure were observed by transmission electron microscope, and the expressions of myocardial HSP70 and HIF-1α were observed by immunohistochemical technology.@*RESULTS@#Myocardial myofibril fracture, mitochondrial cristae and membrane dissolution, and disordered arrangement of Z lines and M lines were observed in electric shock rats. HSP70 and HIF-lα were strong positive expressions in the electric shock death group, significantly compared with the control and postmortem electric shock groups (P < 0.05).@*CONCLUSION@#The expressions of HSP70 and HIF-lα were obviously increased in electric shock death group, which may be used as the diagnostic indicator of electric shock death.

Apoptosis in Lungs and Liver after Crush Injury of Hindlimbs in Rat / 法医学杂志

OBJECTIVE@#To investigate the process of apoptosis in lungs and liver induced by crushing hindlimbs of rat, and study the mechanism of crush injury.@*METHODS@#The rat experimental model of hindlimbs crush injury was established. The cell apoptosis in lungs and liver was detected by TUNEL assay, and the expression of Bax, Bcl-2 and caspase-3 apoptin was examined by immunohistochemistry.@*RESULTS@#Compared with the control group, the partial muscle injury of rat's hindlimbs was more serious with more apoptosis observed in lungs and liver (P < 0.05). The expression of Bax was up-regulated and Bcl-2 was down-regulated, whereas caspase-3 expression was activated (P < 0.05).@*CONCLUSION@#The cell apoptosis has increased significantly in lungs and liver after crush injury of hindlimbs in rat. The correlation factor released during tissue injury may mediate apoptosis process.

Diagnosis and treatment of multiple intracranial aneurysms / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the diagnosis and treatment strategy of multiple intracranial aneurysms (MIA).</p><p><b>METHODS</b>We retrospectively analyzed 96 patients with MIA (234 aneurysms). The rupture site was determined on the basis of computed tomographic and angiographic findings, and the supposed ruptured aneurysm was treated with coiling OR clipping. All the patients' records were reviewed including all computed tomographic scans and angiograms.</p><p><b>RESULTS</b>Twelve patients received conservative treatment, 56 patients were treated by endovascular embolization, and 28 patients received clipping; 44 patients received one-stage treatment, and 4 patients needed a second therapy. In 36 patients, only the ruptured aneurysm was eliminated. The clinical outcomes of these 84 patients evaluated by Glasgow Outcome Scale grades were: absence of deficits in 62 patients, minor deficits in 12 patients, major deficit in 8 patients; death occurred in 2 cases. Thirty patients were available for a 6-month follow-up with DSA, which revealed stable occlusion of the aneurysms in 29 patients and the need of a retreatment due to recanalization in only one patient.</p><p><b>CONCLUSION</b>Correct localization of the rupture aneurysm based on a comprehensive diagnosis is key to MIA treatment. All the aneurysms should be treated in one session whenever possible to protect the patient from rebleeding.</p>

Down-regulation of PTEN expression in kidney and its role in development of diabetic nephropathy in rats / 生理学报

Transforming growth factor-β1 (TGF-β1)-activated phosphoinositide-3-kinase (PI3K)-protein kinase B (PKB/Akt) pathway is intimately related to the development of diabetic nephropathy (DN), which is negatively regulated by phosphatase and tensin homolog deleted on chromosome ten (PTEN). The present study was to investigate the expression of PTEN in the renal tissue of diabetic mellitus (DM) rats and explore its possible effect on development of DN. Sixteen Sprague-Dawley rats were divided into normal control group (n = 8) and diabetic group (n = 8) at random. Streptozotocin injection was used to establish diabetic model. After 12 weeks, the rats were sacrificed to detect relative biochemical parameters and renal index, and to observe the changes of pathomorphology by HE staining as well. In addition, immunohistochemistry staining and Western blotting were employed to detect the protein expression of PTEN, TGF-β1, PI3Kp110α, Akt1, p-Akt1 (Ser(473)), fibronectin (FN) and Collagen IV, respectively. Furthermore, the expression of PTEN mRNA was also examined by RT-PCR. The results indicated that the levels of blood glucose, serum creatinine and urine protein (24 h) were increased remarkably in the diabetic group (P < 0.05) compared with those in the control group. Compared with those in the control group, the protein expressions of TGF-β1, PI3Kp110α, Akt1 in renal tubular epithelium and the expressions of FN and CollagenIV in renal interstitium were increased in the diabetic group (P < 0.05). The expression of PTEN in the diabetic group was significantly reduced than that in the control group (P < 0.05), and the expression of p-Akt1 (Ser(473)) increased remarkably in the diabetic group which had the similar trend to Akt1 (P < 0.05). PTEN mainly located in renal tubular epithelial cells. The expression of PTEN had negative correlation to that of p-Akt1 (Ser(473)). Compared with that in the control group, the expression of PTEN mRNA was decreased remarkably in the diabetic group (P < 0.05). The data suggest that the down-regulation of PTEN in renal tissue of DM rats may promote the PI3K-PKB/Akt pathway over-activated by TGF-β1, which facilitates the initiation and development of DN.

Surgical management of a complex intracranial dural ateriovenous fistula with deep hypothermia circulatory arrest: a case report and literature review / 南方医科大学学报

Hypothermia and cardiopulmonary bypass has been used for difficult lesions of the brain such as giant aneurysms. We reported a case of complex intracranial dural arteriovenous fistula (DAVF) undergoing surgery with deep hypothermic circulatory arrest (DHCA). The advantages and disadvantages of this operation were discussed. This is the first report describing the use of DHCA in the surgical management of complex intracranial DAVF. We also reviewed the literature documenting the treatment of DAVF and the history of deep hypothermia and circulatory arrest in cerebrovascular surgery.

Study on the correlation between syndrome differ classification of knee osteoarthritis and X-ray image / 中国骨伤

<p><b>OBJECTIVE</b>To study the correlation between syndrome differ classification of knee osteoarthritis and X-ray image, so as to provide evidence for clinical diagnosis and treatment.</p><p><b>METHODS</b>From Jun. 2007 to Dec. 2007, 78 patients (108 knees) with knee osteoarthritis were reviewed, including 65 females (89 knees) and 13 males (19 knees), ranging in age from 41 to 77 years. According to the standards for the differentiation of syndrome in the treatment of knee osteoarthritis defined in Principle of Clinical Research for New Traditional Herbs, the patients were divided into three types: Type I, insufficiency of the liver and kidney, with stagnation of tendons and muscles, 43 knees; Type II, insufficiency of the spleen and kidney, with dampness infusion into bone and joints, 26 knees; Type I, deficiency of the liver and kidney, with inter-obstruction of phlegm and stasis 39 knees. Normotopia and lateral plain film of knee joint of weigh loading and in erect position, and patellofemoral Skyline plain flim was taken. Joint space narrow, osteophyte generation, subchondral osteosclerosis and subchondral cystic degeneration were evaluated. All data were analyzed by K independent samples nonparametric test in order to find out the correlation between syndrome differ classification of knee osteoarthritis and X-ray image.</p><p><b>RESULTS</b>It was shown that after K independent samples nonparametric test about syndrome differ classification of knee osteoarthritis and X-ray image: there were significant differences among three types about lateral patella osteophyte, condyles of tibia osteophyte and Type II was the most serious, Type I was secondary, Type II was the lightest. Other index had no obvious difference among the three groups.</p><p><b>CONCLUSION</b>There is certain correlation between syndrome differ classification of knee osteoarthritis and X-ray image. There are significant differences among three types about lateral patella osteophyte, condyles of tibia osteophyte, the Type II is the most serious,Type I is secondary, Type II is the lightest. Consequently, it can be deduced that worse osteophyte is one of accordances of Type II-insufficiency of the spleen and kidney, with dampness infusion into bone and joints. And, the Type II is more serious stage in radiologic manifestation.</p>

Histomorphology of angiogenesis patterns in different areas of human astrocytomas / 南方医科大学学报

<p><b>OBJECTIVE</b>To study angiogenesis patterns in the edematous area and the center of human astrocytomas by histological observation, and to reveal histological basis of vasculogenic mimicry.</p><p><b>METHOD</b>Tissue samples were drawn from the tumor center and the edematous area in 51 patients with human astrocytomas during operation MR and were examined by CD34 endothelial marker periodic acid-Schiff (PAS) dual staining.</p><p><b>RESULTS</b>Vessels or capillaries stained by both PAS and CD34 were found in edematous areas of human astrocytomas. Besides vessels or capillaries stained by both PAS and CD34, vasculogenic mimicries (PAS-positive and CD34-negative tubes containing red blood cells and lined by neoplastic cells), PAS-positive and CD34-negative tubes containing red blood cells and without cells around, PAS-positive and partial CD34-positive vessels or capillaries, and PAS-positive and CD34-negtive vessels or capillaries were detected in the center of tumor of 4 human glioblastomas.</p><p><b>CONCLUSIONS</b>Vasculogenic mimicries in the center of some high-grade astrocytomas may be caused by blood capillary dysplasia, while angiogenesis patterns are vessels or capillaries in the edematus area and the center of most human astrocytomas.</p>

Preparation and identification of the monoclonal antibody specific to hemagglutinin of avian paramyxovirus type 2 / 病毒学报

A hybridoma cell line 1G4A7 secreting monoclonal antibody (McAb) specific to hemagglutinin of avian paramyxovirus type 2 (APMV-2) was developed by fusing the spleen cells of APMV-2 immunized BAlb/c mice with SP2/0 myeloma cells. The immunoglobulin subclass of 1G4A7 was IgG1 with light chain kappa and the affinity constant against APMV-2 was 1.02 X 10(10). Identified by HI and indirect ELISA, the McAb titers in ascities were 10 log 2 and 1 : 10(6) respectively. The McAb did not cross react with the common avian viruses, showing good specificity. There existed obvious differences in antigenitic relationship among APMV-2 viruses analyzed by HI and indirect ELISA using McAb 1G4A7.

Transforming growth factor-β1 and Snail1 mediate tubular epithelial-mesenchymal transition in diabetic rats / 生理学报

The present study was aimed to explore the expressions of transforming growth factor-β1 (TGF-β1) and Snail1 in renal tissues of diabetic rats, and their role in tubular epithelial-mesenchymal transition (TEMT). Induced diabetic rats were randomly divided into 2-, 4-, 8-, 12-, 16-, 20-, 24-week and 16wA, 20wA, 24wA groups. The rats in 16wA, 20wA and 24wA groups were treated with insulin to control blood glucose to the normal level from the 13th week. The age-matched rats were set as controls. Blood glucose, 24-hour urine protein, serum creatinine (Scr), kidney index of rats were measured. PAS staining was used to observe the renal pathological changes. Immunohistochemical staining and (or) Western blot were employed to determine the expressions of TGF-β1, Snail1, E-cadherin, α-smooth muscle actin (α-SMA) and fibronectin (FN) proteins. The expressions of Snail1 and E-cadherin mRNAs in renal cortex were examined by RT-PCR. Blood glucose, 24-hour urine protein, Scr and kidney index increased remarkably in diabetic rats as compared with those in the control groups (P<0.05, P<0.01) and insulin-treated rats (P<0.01). TGF-β1 and Snail1 protein expressions could not be detected by immunohistochemical staining in the normal renal tissues, however, the strongly positive staining was observed in diabetic rat renal tubules. A time-dependent loss of TGF-β1 and Snail1 expressions was detected in the kidney of insulin-treated rats. In diabetic rats tubular α-SMA positive staining was seen at the 16th week. E-cadherin expression was lost in diabetic rats. The expressions of TGF-β1, Snail1 proteins and Snail1 mRNA were significantly up-regulated in diabetic rats, while down-regulated in insulin-treated rats (P<0.01). The expressions of E-cadherin protein and mRNA in the cortex were contrary to the expressions of TGF-β1 and Snail1. Therefore, TGF-β1 and Snail1 are possibly involved in the pathogenesis of TEMT in diabetic nephropathy rats.

p38 MAPK mediates high glucose-induced renal tubular epithelial-mesenchymal transition / 生理学报

The aim of the present study was to investigate the role of p38 MAPK in the renal tubular epithelial-mesenchymal transition (TEMT) induced by high glucose. In in vivo study, the rats were randomly divided into control (C), diabetes mellitus (DM) and insulin-treated DM groups. Immunohistochemical staining and Western blot were employed to determine the expression of p38 MAPK and p-p38 MAPK protein in renal cortex of rats. In in vitro study, primary renal tubular epithelial cells (PTECs) were cultured with normal glucose (5.5 mmol/L), high glucose (20 mmol/L D-glucose), high osmolality (20 mmol/L D-mannitol) and SB202190 (a p38 MAPK inhibitor) plus high glucose respectively for 72 h. The expressions of p38 MAPK, p-p38 MAPK, Snail1, transforming growth factor-beta1 (TGF-beta1), alpha-smooth muscle actin (alpha-SMA) and E-cadherin protein and mRNA were detected by immunocytochemical staining, Western blot and RT-PCR. The p38 MAPK and p-p38 MAPK were specifically upregulated by high glucose in both in vivo and in vitro studies. The p38 MAPK activation was abolished by insulin controlling hyperglycemia to normal level in DM rats and inhibited dramatically by SB202190 in high glucose-cultured PTECs. The protein and mRNA of alpha-SMA were markedly increased in PTECs cultured with high glucose and were 12-fold and 8-fold respectively over that in the normal glucose, which were significantly suppressed by SB202190. SB202190 down-regulated the high glucose-induced Snail1 protein expression in PETCs, and restored partly the depression of E-cadherin protein and mRNA. These results suggest that p38 MAPK mediates high glucose-induced TEMT via transcription factor Snail1.

Transforming growth factor-beta1 and Smad4 signaling pathway down-regulates renal extracellular matrix degradation in diabetic rats / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of transforming growth factor-beta1 (TGF-beta1)/Smad4 pathway in development of renal fibrosis in streptozotocin (STZ)-induced diabetic nephropathy (DN) rats and explore its possible mechanism.</p><p><b>METHODS</b>Male Wistar rats weighing 180-220 g were divided into 5 groups: group A (normal control), group B [diabetes mellitus (DM) 2 weeks], group C (DM 4 weeks), group D (DM 8 weeks), and group E (DM 16 weeks). Except for the normal control group, other groups were induced DM by single injection of STZ (55 mg/kg) respectively. Blood glucose level, serum creatinine, and 24-hour urine protein were examined. Expressions of TGF-beta1 and Smad4 protein and mRNA in kidney were detected using immunohistochemical technique, Western blot, and real-time PCR. mRNA expressions of stromelysin-1 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1), and collagen In in kidney were also detected by real-time PCR.</p><p><b>RESULTS</b>The levels of blood glucose, serum creatinine, and 24-hour urine protein in rats of group B, C, D, and E were higher than those of the control group. With the progression of renal fibrosis, the expressions of TGF-beta1 and Smad4 protein and mRNA in kidney of diabetic rats elevated. In addition, the renal MMP-3 mRNA expression diminished in diabetic rats, while TIMP-1 and collagen III mRNA increased.</p><p><b>CONCLUSIONS</b>In STZ-induced diabetic rats, the TGF-beta1/Smad4 appears to play an important role in renal fibrosis of DN. The increased expression of TGF-beta1 and Smad4 might result in the transcriptional regulation of downstream target genes of TGF-beta1/Smad4 pathway, which contributes to the progression of renal fibrosis in diabetic rats.</p>