[Effects of Zhongfeng capsule on autophagy related proteins expressions of brain tissue in rats with cerebral ischemia/reperfusion injury].

Objective: To investigate the effects of Zhongfeng capsule on the autophagy-related proteins expression in rats with cerebral ischemia/reperfusion injury (CI/ RI), and to explore its neural protection mechanisms of the decoction. Methods: Rat middle cerebral artery ischemia/reperfusion injury model (ischemia for 2 h, reperfusion for 24 h) was prepared by the improved line plug method. Sixty male SD rats were randomly divided into sham operation group, model group, butylphthalide group(0.054 g/kg), Zhongfeng capsule high-dose groups (1.08 g/kg), Zhongfeng capsule middle-dose groups (0.54 g/kg), Zhongfeng capsule low-dose groups (0.27 g/kg), with 10 rats in each group. Rats were treated with Zhongfeng capsule by gavage once a day for 10 days. The rats were sacrificed and the brain tissue was obtained after the experiment in each group. Score neurological deficit was evaluated after 24 h of the last intervention in rat of each group. The pathological changes of brain tissue were observed by HE staining. The serum levels of estradiol (E2) and follicle stimulating hormone (FSH) were determined by ELISA. The expressions of key genes and proteins of PI3K/Akt/Beclin1 signaling pathway in brain tissue were detected by qRT-PCR and Western blot respectively. Results: Compared with the sham operation group, the body weight and protein expressions of p-PI3k and p-Akt in brain tissue of rats were decreased significantly in the model group, while the brain index, neurological deficit score, gene and protein expressions of Beclin1 and LC3 were increased markedly in the model group(P＜0.05 or P＜0.01). In the model group, nerve cells of brain tissue were loosely packed, interstitial edema, triangular in shape, nuclear pyknosis and dark-blue staining were observed. Compared with the model group, the body weight of rats was increased obviously, the neurological deficit score was decreased significantly and the pathological injury of brain tissue was alleviated evidently in high-dose of Zhongfeng capsule group (P＜0.05). The brain index, the gene and protein expressions of Beclin1 and LC3 were decreased apparently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01), while the expressions of p-PI3k and p-Akt in brain tissue were increased evidently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01). Conclusion: Zhongfeng capsule can inhibit autophagy and improve brain neurons lesion of CIRI rats, the mechanism may be related to regulate the expression of Beclin1 and LC3 in PI3K/Akt/Beclin1 signaling pathway.

[Effect of Huangqi decoction on delaying renal cell apoptosis induced by 12C6+ ion irradiation in rats and its mechanism].

Objective: To investigate the molecular protective mechanisms of Huangqi decoction inhibiting the apoptosis of renal cells in the 12C6+ radiation brain model rats. Methods: Fifty Wistar rats were randomly divided into five groups: normal control group, radiation alone model group, Huangqi decoction (high-dose, middle-dose and low-dose ) groups. The normal control group and the radiation alone group were treated with saline10 ml/(kg·d) by gavage, the Huangqi decoction treatment groups were treated with Huangqi decoction at the doses of 4.5, 9 and 18 g/(kg·d) by gavage respectively. After 7 d, except mice in normal control group, the brain of the rats in radiation alone model group, high-dose, middle-dose and low-dose Huangqi decoction group were radiated by 4 Gy 12C6+ ion once. The rats were killed by the femoral artery after irradiation 7 d. The pathomorphism changes of renal tissue were observed by HE, the IL-6 level in serum was detected by ELISA, the gene expressions of Bcl-2, Bax and caspase-3 in renal tissue were assessed by RT-PCR, and the protein expressions of Bcl-2, Bax, caspase-3 and NF-κB in renal tissue were analyzed by immunehistochemical staining. Results: Compared with normal control group, the body weight and kidney index were decreased significantly, the expression of Bcl-2 in renal tissue was decreased significantly, the serum content of IL-6 was increased obviously, and the expressions of Bax, caspase-3 and NF-κB in renal tissue were increased significantly in the radiation alone model group (P＜0.01). The mesangial cells proliferated obviously, interstitial vessels of renal tubules were dilated and congested obviously, the lumen of renal tubules was narrow and irregular in the radiation alone model group. As compared with the radiation alone model group, the body weight and the kidney index were increased obviously in high-dose Huangqi decoction group, the gene and protein expressions of Bcl-2 in renal tissue were increased significantly in Huangqi decoction intervention group(P＜0.05 or P＜0.01). whereas, the protein expressions of Bax and caspase-3 in renal tissue were decreased significantly in middle-dose and high-dose Huangqi decoction group, the serum content of IL-6 was decreased obviously, the gene expressions of Bax and caspase-3 in renal tissue were decreased significantly and the protein expression of NF-κB in renal tissue was decreased significantly in Huangqi decoction intervention group(P＜0.05 or P＜0.01). The proliferation of mesangial cells was improved and the contour of renal tubules was clear in high-dose huangqi decoction group. Conclusion: High-dose of huangqi decoction has protective effect on kidney in rats induced by 12C6+ radiation brain, the mechanism may be related to the regulation of Bcl-2/NF-κB signal pathway.

[Intervention effect of Youguiwan on rats with knee osteoarthritis and its mechanism].

Objective: To study the effects of Youguiwan on the osteoglycin (OGN), osteonectin (ON), fibrinogen 2 (FBN2) of articular cartilage tissue in the model of knee osteoarthritis (KOA). Methods: Sixty SD rats were randomly divided into six groups: sham control group, model group, glucosamine sulfate group, Youguiwan (high-dose, middle-dose and low-dose )group. The modified Hulth method was used to establish KOA models for 6 weeks. The sham control group and the model group were treated with normal saline. The rats in Youguiwan high-dose, middle-dose, low-dose groups were treated with Youguiwan at the doses of 4.8, 2.4, 1.2 g/kg by gavage respectively, and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g/kg. The rats were administrated for 8 weeks according to the dose. After intervening, articular cartilage of rats were obtained, the pathological changes were observed by using HE staining method, and Mankin score was evaluated. The expressions of OGN, ON and FBN2 in articular cartilage were detected by immunohistochemistry. The expression of GSK-3ß in articular cartilage was detected by Western blot. Results: Compared with the sham control group, the Mankin score was obviously increased in the model group, the protein expression of FBN2 was increased significantly, yet the protein expressions of OGN, ON and GSK-3ß were decreased significantly (P＜0.01), articular cartilage was seriously damaged, and chondrocytes were arranged in disorder. Compared with the model group, the Mankin score was declined obviously in the high-dose Youguiwan group, the protein expression of FBN2 was significantly decreased, but the protein expression of GSK-3ß was significantly increased, the protein expressions of OGN and ON were significantly increased in the middle-dose and high-dose Youguiwan group (P＜0.05 or P＜0.01), cartilage structure was tended to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. Conclusion: Youguiwan can significantly improve the articular cartilage degeneration of KOA rats, its mechanism maybe raise OGN and ON protein expression level to promote the ossification and reconstruction of articular cartilage.

[The influence of benazepril and amlodipine on the expression of secretin and somatostatin in spontaneously hypertensive rats].

OBJECTIVES: Investigate the influence of benazepril and amlodipine on the expression of secretin (PZ) and somatostatin (SS) in spontaneously hypertensive rats (SHR). METHODS: Forty-five SHRs (14 weeks old, male) were randomly assigned into 3 groups (n=15):SHR group, Benazepril group (which was given benazepril 0.90 mg·kg-1·d-1) and Amlodipine group (SHRs were given amlodipine 0.45 mg· kg-1·d-1), taking WistarKyoto(WKY) as normal control (n=15), meanwhile, rats in SHR group and WKY group were given the same volume of distilled water. After 8 weeks of intervention, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was detected by enzyme-linked immunoassay and RT-PCR. RESULTS: After 8 weeks of intervention, compared with the WKY group, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was increased significantly in SHR group (P<0. 05). Compared with SHR group, the expression of PZ in duodenum and SS in sinuses ventriculi was decreased significantly in Benazepril group and Amlodipine group (P<0.05). Compared with Benazepril group, in Amlodipine group the expression of PZ mRNA in duodenum and SS mRNA in sinuses ventriculi was decreased more significantly (P<0.05). CONCLUSIONS: The regulation disorder of PZ in duodenum and SS in sinuses ventriculi exists in SHR. The antihypertensive effect of benazepril and amlodipine may be realized by regulating the expression of PZ and SS, while the regulation of amlodipine is more obvious than benazepril.

[The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats].

OBJECTIVES: To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. METHODS: Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (n=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. RESULTS: Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (P<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(P<0.05). Compared with model group, the articular cartilage lesions was light (P<0.05), the Mankin Score was decreased significantly(P<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (P<0.05). CONCLUSIONS: Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.

Co-culture with lung cancer A549 cells promotes the proliferation and migration of mesenchymal stem cells derived from bone marrow.

The initiation and progression of various types of tumors, such as lung neoplasms, are driven by a population of cells with stem cell properties and their microenvironment. Bone marrow mesenchymal stem cells (BM-MSCs) in long-term in vitro culture may exhibit spontaneous changes in stem cell biological properties, including malignant transformations; however, the molecular mechanisms of this have not been fully elucidated. In the present study, a BM-MSC and lung cancer A549 cell co-culture system was utilized to investigate how the tumor microenvironment may spontaneously change the proliferation, migration and differentiation of BM-MSCs. It was demonstrated that the lung cancer A549 microenvironment is able to induce changes in the cell morphology, proliferation, karyotype, cytoskeleton and migration ability of BM-MSCs in vitro. Compared with the control group BM-MSCs, the expression of Ras, phosphorylated-extracellular regulated protein kinases, nuclear factor-κB, P62 and B-cell lymphoma 2 (Bcl-2) proteins in groups of co-cultured BM-MSCs increased significantly (P<0.05) and the expression of P53, Bcl-2 associated X protein and caspase-3 protein decreased significantly (P<0.05). The mechanisms responsible for the changes observed in BM-MSCs may be related to abnormal expression of related genes in the ERK signaling pathway.

[The influences of ultrafiltration and alcohol sedimentation on protective effects of Radix Astragali and Radix Hedyseri against rat's cerebral ischemia].

OBJECTIVE: To investigate the influences of ultrafiltration and alcohol sedimentation on protective effects of Radix Astragali and Radix Hedyseri against rat's cerebral ischemia. METHODS: Using dexamethasone (im.) and ligating common carotid artery, the rat stasis model combined transient cerebral ischemia was established to evaluate the effects of the ultrafiltration and alcohol sedimentation through detecting antioxidant system and other indexes in brain tissue. RESULTS: The results showed that the 6 g/kg water extract(crude drug), ultrafiltration and alcohol sedimentation of Radix Astragali and Radix Hedyseri could upgrade adenosine-triphosphate (ATP), superoxide dismutase (SOD) and catalase (CAT), and degrade malondialdehyde(MDA) and water content of brain tissue in rat stasis model combined transient cerebral ischemia, the water extract and ultrafiltration of them could degrade lactic acid (LD) of brain tissue, and the effects of alcohol sedimentation of Radix Astragali and Radix Hedyseri become weaker than water extract of them. CONCLUSION: The water extract, ultrafiltration and alcohol sedimentation of Radix Astragali and Radix Hedyseri have some protective effects on cerebral ischemia in rats, the effective differences of the extract through the same extraction method are not remarkable, and alcohol precipitation method has obvious influences effect on Radix Astragali and Radix Hedyseri.

[Effect of JTS caps. treating cerebral ischemia on metabolism and antioxidant system in cerebral ischemia rat].

OBJECTIVE: To study the effects of JTS, Traditional Chinese Medicine caps. treating cerebral ischemia on metabolism and antioxidant system in cerebral ischemia rat. METHODS: I.m. dexamethasone and ligating common carotid artery, the model of cerebral ischemia rats was established to investigate the effects of JTS caps. and its mechanisms through detecting substance metabolism, energy metabolism and antioxidant system. RESULTS: JTS caps. (1.78 - 3.56 g/kg) could upgrade glucose (Glu), total amino acids (T-AA), ATP, Na(+)-K(+) -ATPase, superoxide dismutase (SOD) and catalase (CAT) of brain tissue and degrade lactic acid (LD), malondialdehyde (MDA) and water content of brain tissue in cerebral ischemia rat (P < 0.05, 0.01). JTS caps. (3.56 g/kg) could also depress extenuation of rat's body weight. CONCLUSION: JTS caps. has some protections against the cerebral ischemia in rats, and one of the mechanisms may be improving the metabolism and antioxidant system.

Diagnostic value of ADC and rADC of diffusion weighted imaging in malignant breast lesions / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the value of diffusion weighted imaging (DWI) in the diagnosis of patients with breast diseases.</p><p><b>METHODS</b>Fifty-three consecutive patients were scanned with GE signa HDx 1.5 T magnetic resonance system equipped with 8-channel breast coil. DWI was scanned by SE-EPI sequence in b values of 500 s/mm(2) and 800 s/mm(2), respectively. The apparent diffusion coefficients (ADC) of these lesions were measured. The mean apparent diffusion coefficients (ADC) of these lesions were calculated in b values of 500 s/mm(2) and 800 s/mm(2), respectively. These lesions' ADC value (rADC) was counted respectively and the result of the rADC was equal to the lesion's ADC divided by the ADC of the ipsilateral normal breast tissue. Threshold of ADC and rADC for differential diagnosis was acquired by ROC (receiver operating characteristic curve) analysis. Different imaging technologies were evaluated emphasizing their sensitivity, specificity and accuracy.</p><p><b>RESULTS</b>Sixty-six lesions of 53 cases were confirmed by pathology, including 39 malignant lesions and 27 benign lesions. (1) b = 500 s/mm(2), the threshold of ADC value was 1.435 x 10(-3) mm(2)/s, with a sensitivity of 82.1% and a specificity of 81.5%. The threshold of rADC value was 0.62, with a sensitivity of 76.9% and a specificity of 100%. (2) b = 800 s/mm(2), the threshold of ADC value was 1.295 x 10(-3) mm(2)/s, with a sensitivity of 79.5% and a specificity of 81.5%. The threshold of rADC value was 0.71, with a sensitivity of 89.7% and specificity of 88.9%. (3) The area under the ROC curve was increased for the four diagnostic indicators (ADC(500), ADC(800), rADC(500), rADC(800)).</p><p><b>CONCLUSION</b>DWI spends short time, and it doesn't need contrast material. ADC value and rADC value have a high sensitivity and specificity as a diagnostic indicator. DWI is helpful in improving the specificity of MR and may become one of valuable conventional procedures for breast tumor diagnosis.</p>

Imaging features and clinicopathological manifestations of solitary fibrous tumors / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the imaging features, clinical manifestations and pathological characteristics of solitary fibrous tumors (SFT).</p><p><b>METHODS</b>The clinicopathological manifestations and medical imaging findings were analyzed retrospectively in 27 patients with surgically confirmed SFT.</p><p><b>RESULTS</b>The SFTs originated from different parts of the body, including 18 in the chest, 4 in the abdomen, 1 in the lumboscral area, 3 in the pelvis, and 1 in the left shoulder. Twenty-three cases were found by CT scan, among which there were 16 benign diseases, presented with well-defined round or elliptic margins, with homogeneous attenuation and clearly surrounding; 6 malignant cases with unclear demarcations, invasive surrounding, heterogeneous attenuation due to calcification and/or irregular necrosis, and 1 junctional case with well-defined margins, which was enlarged during follow-up. There were 4 SFTs scanned by MRI with clear margin and homogeneous or heterogeneous signal intensity. All of the 4 cases were isointense or hyperintense to muscle on T1-weighted images, and were hyperintense on the T2-weighted images. All tumors showed heterogeneously intense enhancement with geographic pattern. Immunohistochemical staining showed that CD34-positive was 81.5%, vimentin (100.0%), CD99 (100.0%) and bcl-2 (96.3%), as well as negative CK (100.0%) and S-100 (96.3%).</p><p><b>CONCLUSION</b>The location of SFT is varying. Though its clinical manifestations vary, the diagnosis is depended on pathology and immunohistochemistry. There are certain specific features related to SFTs on CT or MRI. These imaging techniques may serve to provide helpful information as to the location and vicinal anatomic structure of the tumor, which is of substantial importance for planning surgery.</p>