This study was aimed to observe the influence on migration ability of human umbilical vein endothelial cell (HUVEC) in vitro by couplet medicines of reinforcing qi and activating blood, and initially screen medicines with relative obvious promoting or inhibiting effect on migration ability of HUVEC. The model of HUVEC cultured in vitro was established. The Paeonia, Ligusticum chuanxiong, Cortex moutan, Salvia, curcuma zedoaria, Sparganium, Astragalus, and ginseng were combined in pairs with the proportion of 1:2, 1:1, 2:1. There were 13 couplet medicines of reinforcing qi and activating blood. Serum pharmacological method was used to prepare medicated serum of the couplet medicines of reinforcing qi and activating blood. Scratch method was used to detect the effect of medicated serum of these couplet medicines of reinforcing qi and activating blood and activate blood herbs on the migration a-bility of HUVEC (with the density of 5í105/mL) after 24 hours. The results showed that compared with the blood serum of the blank group, the Cortex moutan group, Ligusticum chuanxiong group, Paeonia group, Salvia and Astra-galus (1:2) group, Salvia and Astragalus (1:1) group, Ligusticum chuanxiong and Astragalus (1:2) group had obvious promoting effect on the migration ability of VEC (P < 0.05 or P < 0.01). The Sparganium group, curcuma zedoaria group, Cortex moutan group, curcuma zedoaria and Astragalus (2:1) group, Sparganium and ginseng (2:1) group, Spar-ganium and Astragalus (1:1) group had obvious inhibiting effect on the migration ability of VEC (P< 0.05 or P<0.01). It was concluded that different compatibility of medicines of reinforcing qi and activating blood and activate blood herbs had different promoting or inhibiting effect on migration ability of HUVEC. It may be related to the mechanism of action of these drugs on promoting or inhibiting angiogenesis at the cellular level.

This study was aimed to observe the influence of qi-reinforcing and blood-activating(QRBA) herbs onan-giogenesis of the myocardial microvascular, SDF-1 and CXCR4 protein expression as well as mRNA expression in the infarcted myocardium edge area of acute myocardial infarction (AMI) ratmodel. The AMI rat model was estab-lished. The immunohistochemical staining method was used in the detection of vWF protein expression in the myocar-dial tissues. The MVC account was recorded. The SDF-1 factor and its specific receptor factor CXCR4 were detected by the western blot and realtime-PCR technique in the infarcted myocardium edge area of rats from each group. The results showed that in the new generated microvessels which were staining marked by the vWF factor can be seen in infarcted myocardium edge area of rats from the sham-operated group, model group, each medication group. The new generated microvessels in the myocardium of rats in the sham-operated group were not obvious. Small amount of new generated microvessels can be seen in rats from the model group. More new generated microvessels can be seen in rats from each medication group. The comparison between the model group and the sham-operated group showed sta-tistical difference (P<0.05). The comparison between each medication group and the model group showed statistical difference(P<0.05 or P<0.01). Compared with the sham-operated group, SDF-1, CXCR4 and mRNA expression were obviously increased in the myocardium of rats in the model group (P<0.05 or P<0.01). Compared with the model group, SDF-1, CXCR4 protein and mRNA expression were obviously increased in the myocardium of rats from each medication group (P<0.05 or P<0.01). It was concluded that herbs such as Salvia, couplet herbs of Salvia and Astra-galushad stimulation effectonangiogenesis. Mechanism of these drugs in angiogenesismay be through the promotion of SDF-1 and CXCR4 protein as well as mRNA express.

Objective To analyze the protein expression and subcellular distribution of mechanogrowth factor (MGF) in ostcoblasts under stretch stimulation. Methods Cyclic stretching was applied to osteohlasts by a mechanical stretching device. The whole-cell proteins were extracted from controlled and stretched osteoblasts for detecting the protcin expression level of MGF by Western blot and observing the intracellular distribution of MGF by fluorescent immunocytological method. Results Western blot showed significant increase of expression of MGF in osteoblasts under stimulation of cyclic stretching. The level of protein was increased by four folds after 12-hour stretching of osteohlasts, and then declined sharply. Immunofluorescence analysis showed that MGF was mainly distributed in the nuclei of osteoblasts. ConcinsionsUnder the cyclic stimulation, the expression of MGF reaches a short period of peak in osteoblasts, which may be related to the injury of osteoblasts caused by stretching. MGF is mainly distributed in the nuclei of osteoblasts, indicating that MGF may contain nuclear localization signal and modulate the expression of relative genes.

ObjectiveTo investigate the clinical features and prognosis of children with rotavirus gastroenteritis and convulsion.MethodsClinical data of children with rotavirus gastroenteritis hospitalized from January 2010 to December 2013 were retrospectively analyzed. Subjects were divided into the seizure group and no seizure group according to the presence of seizure in the course and compared between the two groups.ResultsThere were no signiifcant differences in sex, age, and the average duration of hospitalization between two groups (allP>0.05). The family history, history of seizures, the levels of serum sodium, calcium, lactate, standard bicarbonate concentration (SB), actual bicarbonate concentration (AB), carbon dioxide content (TCO2) and pH were statistically signiifcant between two groups (allP>0.05). During the follow-up period (outpatient telephone follow-up), the recurrence of seizure in two groups was signiifcant different (P<0.05) and only one (0.54%) child in seizure group developed epilepsy.ConclusionThis study showed that rotavirus gastroenteritis with convulsion is a benign clinical course.

Mechano-growth factor (MGF) is one of IGF-1 isoforms. MGF is mechanosensitive and has important functions in muscle hypertrophy, regeneration and nerve injury recovery. In this study, MGF cDNA (330 bp) was cloned from stretched osteoblasts by RT-PCR. In order to avoid prolin residue inhibiting enterokinase cleavage, 9bp of MGF cDNA 5' end sequence was truncated by primer, then the obtained truncated MGF (des(1-3)MGF) cDNA (321 bp) was subcloned in pET32a(+) vector to construct a prokaryotic recombination expression plasmid. Trx/des(1-3)MGF fusion protein, existing in forms of solution, was expressed in transformed Escherichia coli strain BL21(DE3) by IPTG induction at 30 degres C. The supernatant of cell lysates was subjected to ion exchange chromatography and Ni2+ metal affinity chromatography, and the fusion protein was obtained with the purity over 95%. After the fusion protein was cleaved by enterokinase, Trx and des(1-3)MGF was isolated by reverse-phase HPLC. Through these procedures, des(1-3) MGF was obtained with the purity of 98%. The protein molecular mass was conformity to the theoretical value by SDS-PAGE and mass spectrometry analysis. The purified des(1-3)MGF was incubated with MC3T3-E1 for cell proliferation and migration assays. The results show that des(1-3)MGF exhibited more facilitative effects on proliferation and migration of MC3T3-E1 than that of des(1-3)IGF-1.
Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Insulin-Like Growth Factor I ; Osteoblasts ; metabolism ; Protein Isoforms ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; STAT5 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

This is an experimental study in the realm of physiology inquiring about the effect of pulsatile fluid flow shear stress on the proliferation, differentiation and functions of osteoblasts;the objective is to validate the important effect of fluid flow shear stress on the mechanics adaptability of bone tissue. The osteoblasts derived from Wistar rat's calvaria were exposed to fluid shear stress 5, 10, 20 and 30 mN/cm2 for 3, 6, 9, 12, 24, 36h respectively in the flow chamber. The ability of proliferation, alkaline phosphatase (ALP) activity and extracellular calcium secretion of osteoblasts were assessed. The results showed that fluid flow shear stress at 5, and 10 mN/cm2 increased the proliferation, but at 20 and 30 N/cm2, the shear stress inhibited the proliferation. The shear stress at 5, 10, 20 mN/cm2 increased the ALP activity and extracellular calcium secretion of osteoblasts, and advanced the time of the peak value of ALP activity during the experiment period, but the shear stress at 30 mN/cm2 decreased ALP activity. So osteoblasts responded rapidly to shear stress; the proliferation, differentiation and mineralization of cells were regulated in the presence of some shear stress; and such regulation exhibited a pattern of dependence on the mN/cm2 level of shear stress.
Alkaline Phosphatase ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Mechanotransduction, Cellular ; physiology ; Osteoblasts ; cytology ; enzymology ; Pulsatile Flow ; Rats ; Rats, Wistar ; Shear Strength ; Skull ; cytology ; Stress, Mechanical

With the use of a cyclic strain unit, the proliferation and gene expression of IGF-1 in the rat osteoblasts that underwent mechanical strain were studied. The cells were subjected to 15% elongation at frequency 20 cycles/min for different loading time. Under the action of different loading time, the relative proliferation index of the rat osteoblasts was the biggest of all when loading time was 12h; during the course, the expression of IGF-1 mRNA increased significantly, and then gradually tended toward 1 with the increase of the loading time. These results demonstrate that osteoblasts respond to the mechanical forces which may regulate the activities of osteoblasts indirectly by promoting the autocrine effect of IGF-1. Loaded osteoblasts can adjust and adapt themselves to new mechanical stimulation, and hence maintain a new state of equilibrium.
Animals ; Animals, Newborn ; Cell Proliferation ; Cells, Cultured ; Gene Expression ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Skull ; cytology ; Stress, Mechanical

OBJECTIVE To explore whether pyridoxine can protect acute kidney injury caused by cisplatin and to analyze its specific mechanism. METHODS After establishing an in vitro model of cisplatin-induced damage in HK-2 cells, different concentrations of pyridoxine were administered and cell survival was detected using SRB, the expression of apoptosis-related and antioxidant proteins were detected by Western blotting, and the activity of reactive oxygen species(ROS) and superoxide dismutase(SOD) were detected by kits. A cisplatin-induced mouse kidney injury model was further established, and the serum urea nitrogen level was detected after the administration of 40 mg·kg–1 pyridoxine treatment, the results of hematoxylin-eosin staining in renal tissue were analyzed, and the expression of NRF2 was detected by Western blotting. RESULTS Pyridoxine could protect kidney injury caused by cisplatin in HK-2 cells and mouse in vivo. In HK-2 cells, pyridoxine down-regulated ROS level, up-regulated SOD enzyme activity, and up-regulated the expression of NRF2 and its downstream antioxidant-related gene HO-1. Pyridoxine significantly reduced the level of serum urea nitrogen, repaired kidney tissue damage, and up-regulated the expression of NRF2 in kidney injury mice. CONCLUSION Pyridoxine protects against cisplatin-induced kidney injury through enhancing the level of anti-oxidative stress.

OBJECTIVETo evaluate treatment modalities and prognosis differentiated thyroid carcinoma (DTC) with tracheal invasion.

METHODSClinical data were reviewed and analyzed in 50 patients treated for DTC with tracheal invasion between January 1990 and June 1998. The different surgical modalities were applied according to the extent of tracheal invasion: shave excision (20 cases), tracheal sleeve resection or tracheal partial resection (23 cases), total laryngectomy or laryngeal closure surgery (7 cases). Thirty-eight cases received postoperative (131)I therapy. Survival rate was evaluated using the Kaplan-Meier analysis.

RESULTSThe 5-, 10- and 15-year survival rates of all the cases were 90.0%, 74.0% and 56.0%, respectively. The 5-, 10- and 15-year survival rates were 94.7%, 81.6% and 65.8% respectively in 38 cases with postoperative (131)I therapy and were 75.0%, 50.0% and 25.0% respectively in 12 cases without postoperative (131)I therapy, with statistically significant differences in 5-, 10- or 15-year survival rates between the patients of two groups.

CONCLUSIONSThe tumors can be resected radically by corresponding surgery based on the extent of tracheal invasion. Postoperative (131)I therapy can enhance the survival rate of the patients with differentiated thyroid carcinoma involving in trachea.

Adenocarcinoma ; Humans ; Kaplan-Meier Estimate ; Laryngectomy ; Neoplasm Invasiveness ; Prognosis ; Retrospective Studies ; Survival Rate ; Thyroid Neoplasms ; diagnosis ; therapy ; Trachea ; Tracheal Neoplasms ; diagnosis ; secondary ; therapy