Objective To investigate the proper inner control genes suitable for mRNA expression level comparison of aging rat tissues.Methods Real-time reverse transcription PCR was used to examine in aging rat tissues the expression level of G3pd(glyceraldehyde-3-phosphate dehydrogenase),ACTB(?-actin),H3f3b(H3 histone,family 3B),Arbp(acidic ribosomal phosphoprotein P0)and 18S(18S ribosomal RNA).Results The most stably expressed housekeeping gene in aging rat kidney was ACTB,in heart and lung G3pd showed the minimum variation;Arbp expression was the most stable one in different tissues.Conclusion For aging rat intra-tissue mRNA normalization at least two housekeeping genes should be used: one is the ribosomal RNA gene 18S and another one is Arbp.

Objective To explore the value of dexamethasone(DEX) for neuronal cell injury and death by observing the effect of DEX on excitatory amino acid(EAA) and monoamine neurotransmitter in cerebral tissue of neonatal rat with hypoxia-ischemia.Methods Hypoxic-ischemic neonatal rat models were established,the levels of EAA and monoamine neurotransmitter in cerebral tissue were analyzed by using capillary electrophoresis and fluorospectrophotometry method.The rats were divided into 4 groups: small dose DEX group pre-treated with DEX(0.5 mg/kg) prior to hypoxia-ischemia,large dose DEX group pre-treated with DEX(10 mg/kg) prior to hypoxia-ischemia,HIE group and shamful operation group.Results The levels of EAA and monoamine neurotransmitter contents in HIE group were significantly higher than those in shamful operation group(P0.05).EAA contents of large dose DEX group greatly decreased compared with HIE group (P

Objective To evaluate the significance of ST/TP relative value in electrocardiogram in diagnosis of hypocalcemia.Methods ST/TP relative value in electrocardiogram in 61 children with hypocalcemia and 31 normal children were detected by NIHOH KOHDEN CardiofaxQ ECG-9130P electrocardiogram machine designed by Japanese futian company.Calcium ion was checked out by clinic electrolyte equipment.Results There was significant difference of ST/TP relative value between patients with hypocalcemia and normal children(P

Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of human spermatogenetic mechanism.

Objective To evaluate influence of Azithromycin on breath hydrogen test(BHT) in acute upper respiratory infection(AURI) in Children.Methods Fasting BHT and lacunose BHT were respectively performed by using HD-1 breath hydrogen detector for 38 inpatients before and after receiving Azithromycin.Fasting BHT of 115 healthy controls were determined.Results After Azithromycin was given,Fasting BHT was lower than that before Azithromycin used(P

OBJECTIVE: To determine the effect of different tidal volume(Vt) ventilation on apoptosis and the expression of Bax and Bcl-2 in rat lungs. METHODS: Twenty-four healthy SD rats were randomly divided into 4 groups: a control group, a low Vt ventilation group (LV), a middle Vt ventilation group (MV), and a high Vt ventilation group (HV). Rats were subjected to different tidal volumes (10,20,and 40 mL/kg) for 2 h except the control group, which kept their own breath. We determined the lung histopathology score, W/D ratio and WBC in bronchoalceolar lavage fluid (BALF) to evaluate the lung injury and examine the apoptotic cell death, Bax and Bcl-2 protein expression by using TUNEL technique and immunohistochemistry 24 h after the operation. RESULTS: Compared with the control group, MV and HV increased lung histopathology score, W/D ratio, WBC in BALF, apoptosis index (AI )and Bax protein expression, but decreased Bcl-2 protein expression (P<0.05). These changes showed no significant difference between the control group and the low Vt ventilation group (P>0.05). CONCLUSION Low Vt ventilation contributed little to apoptosis. Higher Vt ventilation can improve Bax while inhibit Bcl-2 expression to aggravate apoptosis in rat lungs.
Animals ; Apoptosis ; physiology ; Female ; Lung ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiration, Artificial ; adverse effects ; methods ; Tidal Volume ; physiology ; bcl-2-Associated X Protein ; metabolism

Objective To evaluate the histological response and the clinical value in laparescopic colorectal surgery combined with preoperative regional intra-arterial chemotherapy(PRAC). Methods In cases of rectal cancer and fiver metastases selective regional intra-arterial chemotherapy and iodinated oil embolism was carried out in 23 cases of colorectal carcinoma. After 1 to 11 days laparoseopic radical resection was done, specimens were sent for histopathological examination. We analyzed the correlation between tumor differentiation and TNM stage, compared the effect of PRAC with PRAC + embolism by the criteria of histological response of chemotherapy. χ2-test was used to compare interclass correlation.Results The histological effect in the 23 cases of PRAC level 0 was in 2 cases, level Ⅰ in 7 cases, level Ⅱ in 10 cases, and level Ⅲ in 2 cases. The overall effective rate was 91% (21/23). In the 15 cases with lymph node metastases, the effective rate was 87% (13/15). There was no significant statistic correlation between tumor differentiation or TNM stage and histological response. PRAC associated embolism had a better histological response compared to PRAC alone. Conclusions Preoperative regional intra-arterial chemotherapy had marked therapeutic effect on histological response to the colorectal carcinoma patients of various tumor differentiations and TNM stage, especially combining with the embolism to rectal cancer could improve the efficacy.

OBJECTIVE: To explore the effect of lentivirus-mediated BMP-2 overexpression plasmid transfection into bone marrow mesenchymal stem cells and silk fibroin scaffold on osteoblast transformation. METHODS: The lentivirus BMP-2 overexpression vector was constructed, bone marrow mesenchymal stem cells were cultured, and the combined culture system of nuclear scaffolds was constructed. Alizarin red staining and alkaline phosphatase staining were used to detect the osteogenic transformation of bone marrow mesenchymal stem cells in vitro. Ten New Zealand white rabbits, weighing 3.2 to 4.5 kg(averaging 3.9 kg), aged (2.89±0.45) years old, were selected to construct the rabbit tibial defect model by drilling a conical tibial defect (5 mm in length, 2 mm in width and 3 mm in depth) with an oral drill. The repair of the tibial defect in the animal model was observed by HE staining. The experimental group was implanted with silk fibroin scaffold + BMP-2 overexpression vector bone marrow mesenchymal stem cell complex, while the negative control group was implanted with silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cell complex. RESULTS: Compared with the control group(silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cells), the number of adherent cells on the surface of the scaffold in the experimental group(silk fibroin scaffold+transfected BMP-2 overexpression vector BMP-2 complex) increased significantly. Compared with the control group, the ECM secretion in the experimental group increased significantly. EDX analysis showed that the content of calcium ion was 0.22% in the control group and 0.86% in the experimental group, which showed that the ability of inducing calcium ion formation in the experimental group was stronger than that in the control group. Alizarin red staining of calcium nodules showed that there was no obvious change in the naked eye of the control group, and a small amount of calcium nodules could be seen under the microscope. In the experimental group, obvious red area staining was observed by naked eye, and a large number of calcium nodules were observed by microscopy. The results of alkaline phosphatase staining showed that there was no obvious change in the naked eye of the control group, and no obvious change in the microscopic observation. In the experimental group, purple area staining was observed by naked eyes, and ALP staining was strongly positive by microscopy. The combined culture system of silk fibroin scaffold and bone marrow mesenchymal stem cells can repair cartilage defects. The repair effect of BMP-2 bone marrow mesenchymal stem cells after transfection is obviously better than that of non-transfection group. HE staining showed that inflammatory cells decreased and scaffolds disappeared slightly in the control group. In the experimental group, inflammatory cells were significantly reduced, scaffolds disappeared and angiogenesis was observed. CONCLUSIONS Lentivirus-mediated BMP-2 overexpression plasmid can promote BMSC to differentiate into osteocytes and secrete more extracellular matrix containing Ca²⁺ to promote bone defect repair.
Animals ; Bone Marrow Cells ; Bone Morphogenetic Protein 2 ; Cells, Cultured ; Fibroins ; Lentivirus ; Mesenchymal Stem Cells ; Osteoblasts ; Osteogenesis ; Plasmids ; Rabbits ; Transfection

OBJECTIVETo culture and amplify the young rabbit's bone marrow stromal cells (BMSCs) in vitro, and to observe the effect of hypothermia on the cells' growing behavior and biological function.

METHODSBMSCs were acquired from the rabbit' tibia bone marrow and induced to mature osteoblasts in vitro. The cultured cells growing well in vitro were preserved in liquid nitrogen. The anabiotic cells having cryopreserved for 1 week were chosen as the experimental group, and the routine 7th generation as the control group. Their biological function in comparion by the examination of morphological changes, cells' proliferation ability, colone forming ratio, synthesis ability of ALP and protein, mineralized nodes forming ability were observed.

RESULTSAs contrast to the control groups, the anabiotic cells also grew and proliferated well in vitro except a little more slowly than before. They had the similar general shape in all the time segments, but a little differences in cells' ultrastructure. The experimental groups also had the typical characters of mature osteoblasts, and high abilities of the synthesis of ALP and proteins. The statistic data showed that these two groups had no significant difference (P > 0.05).

CONCLUSIONThe cryopreserved osteoblasts had the same biological functions and the similar growing behaviors as before. These results suggest that it is practical to use the cryopreserved osteoblasts for further study on bone tissue engineering.

Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; In Vitro Techniques ; Osteoblasts ; Rabbits ; Tissue Engineering

OBJECTIVETo investigate the essential role and mechanism of TRPC6 gene in the development of gastric cancer.

METHODSThe expression of TRPC6 protein was assessed in gastric cancer tissues and normal tissues adjacent to the cancer from 30 patients with gastric cancer. The inhibiting effect of TRPC6 activity on cell growth, cell cycle of a human gastric cancer cell line AGS cells, tumor progression and development of xenografted human gastric cancer in a mouse model was tested using dominant-negative mutant TRPC6 (DNC6). The survival of mice bearing xenografted tumors in the GFP and DNC6 was compared using Kaplan-Meier analysis. All statistical tests were two-sided.

RESULTSThe TRPC6 protein in the tumor tissues and para-tumor tissues was (21.60 ± 8.32)% versus (7.14 ± 2.24)%. After transfection of DNC6 virus for 24 hours, 48 hours, 72 hours and 96 hours, the growth inhibition rates of gastric cancer cells were (36.90 ± 1.13)%, (44.06 ± 2.17)%, (52.12 ± 2.76)% and (50.89 ± 1.97)%, respectively. The clone formation rates of control group and DNC6 group were (14.70 ± 3.00)% versus (43.80 ± 7.00)%. After transfection with DNC6 virus for 0, 24, 36 and 48 hours, the G(2)/M phase arrest was (20.34 ± 1.98)%, (24.31 ± 2.37)%, (27.70 ± 2.36)%, (35.10 ± 3.0)% in the DNC6 group and (18.40 ± 2.01)%, (18.0% ± 1.72)%, (17.50 ± 1.74)%, (16.80 ± 1.71)% in the control group, respectively. Inhibition of TRPC6 activity also reduced the subcutaneous tumor volume in the mouse models with xenografted human tumors (P < 0.05).

CONCLUSIONIn the preclinical models tested, TRPC6 channels are essential for gastric cancer development via regulation of G(2)/M phase transition.

Adenoviridae ; genetics ; Animals ; CDC2 Protein Kinase ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin B ; metabolism ; Cyclin-Dependent Kinases ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Recombinant Proteins ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; TRPC Cation Channels ; metabolism ; TRPC6 Cation Channel ; Transfection ; Tumor Burden