Objective To study the effect of age and sex on circadian rhythms of renin-angiotensin system(RAS) .Methods 96 SPRD rats were divided into three groups.The levels of renin(Ren)activity,angiotensin Ⅱ(Ang Ⅱ)and angiotensin-converting-enzyme(ACE)in plasma or serum were respectively determined.Results The plasma or serum levels of renin,ACE,and angiotensin-Ⅱ in adult male group(D group)were significantly increased in comparison with elderly male group(M group)and elderly female group(F group).There were not significantly different between F group and M group in the levels of Ren activity,AngⅡ and ACE.The three substances presented typical circadian rhythms in three groups(P

Animals ; Hypoxia ; classification ; metabolism ; Male ; Oxygen Consumption ; Rats ; Rats, Sprague-Dawley

Objective To identify changes in the occurrence of endemic fluorosis in order to provide scientific basis for making countermeasures. Methods Five villages from 14 counties of mild, moderate and severe fluorosis affected areas were selected by stratified cluster sampling every year in the whole province during 2006 - 2010. Water and urinary fluorine were determined by ion selective electrode method(GB/T 8538-1995); dental fluorosis of children 8-12 years old was diagnosed with Dean method; skeletal fluorosis was diagnosed according to "clinical indexing standards of endemic skeletal fluorosis "(GB 16396-1996), between 2006 and 2008, and "clinical diagnosis standard of endemic skeletal fluorosis"(WS 192-2008) between 2009 and 2010. Results A total of 25 diseased villages were surveyed, 14 with water sources changed, covered a resident population of 8005 people, beneficiary population 7154, and accounting for 89.37% of the resident population; not changed villages 11. In accordance with the "State drinking water health standards", in the 14 changed villages the fluoride in drinking water was qualified (≤ 1.20 mg/L), there were 3 schools whose water fluorine content exceeded the standard; among the 11 villages that did not change water sources 7 drinking water samples fluorine content exceeded the standard. Of the 8 to 12 years old children in villages with changed water sources, 363 of them were checked and 142 dental fluorosis were found, the detection rate of dental fluorosis was 39.12% (142/363); in villages with water sources not changed, 303 children were checked, the detection rate of dental fluorosis was 43.89%(133/303). Of sixteen and elder adults in water source changed villages, 6424 people were checked and 403 skeletal fluorosis were found, skeletal fluorosis detection rate was 6.27% (403/6424); 3572 people were checked in not changed villages, the detection rate of skeletal fluorosis was 13.89%(496/3572). In water sources changed areas, geometric mean of urinary fluoride was in the normal reference value(WS/T 256-2005, 1.40 mg/L)or less. Conclusions Endemic fluorosis is decreased in water improved areas, but in unimproved areas the disease is still severe, and control of endemic fluorosis is still an arduous task.

OBJECTIVETo study the clinical efficacy of the laparoscopic surgery in combination of Quyu Jiedu Recipe (QYJDR) on endometriosis (EMs)-related infertility patients.

METHODS130 EMs-related infertility patients of blood stasis and toxin accumulation syndrome diagnosed and treated by laparoscopic surgery were randomly assigned to two groups, i.e., the test group (75 cases) and the control group (55 cases). All patients received routine progesterone treatment after laparoscopic surgery. QYJDR was given to patients in the test group after laparoscopic surgery, lasting for twelve months. The pregnancy rate within one year after laparoscopic surgery were compared between the two groups. The Chinese medicine syndrome integrals, serum endometrium antibody (EMAb) and cancer antigen 125 (CA125) levels were compared between before drug therapy and three months after drug therapy.

RESULTSThe pregnancy rate within one year after laparoscopic surgery was higher in the test group than in the control group [61.33% (46/75) vs 45.45% (25/55)], showing significant difference (P<0.05). The Chinese medicine syndrome integral was (16.07 +/- 6.77) score and (7.25 +/- 3.27) score before and 3 months after drug therapy in the test group, while it was (15.92 +/- 7.51) score and (12.73 +/- 6.12) score in the control group. The Chinese medicine syndrome integral after drug therapy was lower than that before drug therapy in the same group. Besides, the integral difference between before and after drug therapy was higher in the test group than in the control group [(9.12 +/- 6.16) score vs (3.48 +/- 2.06) score)], showing statistical difference (P<0.05). After three-month drug therapy, the serum EMAb negative conversion rate was obviously higher in the test group than in the control group (44.44% vs 15.62%), showing statistical difference (P<0.05). The serum CA125 level was (31.88 +/- 15.78) U/mL before drug therapy and (18.82 +/- 10.08) U/mL three months after drug therapy in the test group, while it was (30.63 +/- 19.28) U/mL and (18.05 +/- 11.20) U/mL respectively in the control group. It was lowered after drug therapy in the two groups with statistical difference (P<0.05). There was no statistical difference between the two groups after drug therapy (P>0.05).

CONCLUSIONSThe laparoscopic surgery in combination of QYJDR could effectively improve clinical symptoms of EMs patients of blood stasis and toxin accumulation syndrome, promote negative conversion of EMAb, lower serum CA125 levels, and elevate the clinical pregnancy rate. QYJDR is an effective formula in treatment of EMs-related infertility.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; complications ; therapy ; Female ; Humans ; Infertility, Female ; etiology ; therapy ; Integrative Medicine ; Laparoscopy ; Phytotherapy ; Treatment Outcome

Objective To investigate the expression and significance of Caspase-3 and Survivin at different stages of human hemangioma in children.Methods Fifty-five specimens of hemangioma tissue excised in operation and 8 normal skins removed in operation were harvested from People's Hospital of Gansu Province between Jan.2000 and Dec.2005.The pathologic diagnosis was divided into 3 groups according to Mulliken's standard under microscope:proliferated phase group(n=31),degenerated phase group(n=24),and control group(n=8).All these specimens were examined by immunohisto for Caspase-3 and Survivin expression.Results Caspase-3 positive rate in the proliferative phase and involuting phase were 35.5% and 79.2%,respectively.The positive rate in involuting phase was higher than that that in proliferative phase,the difference between the 2 groups was significant(P=0.045 9).A significant difference was not found between the proliferative phase and normal skin tissue(P=0.057 3).Survivin positive rate in the proliferating and involuting phase were 77.4% and 45.8%,respectively.The positive rate in proliferative phase was higher than that in involuting phase,the difference between the 2 groups was significant(P=0.008 5),the difference in involuting phase and normal skin tissue was not significant(P=0.059 3).Conclusions High level of Caspase-3 expression in vascular endothelial cells in involuting phase in contrast to that in proliferative phase,which indicates that Caspase-3 may play a positive role in the apoptosis of vascular endothelial cells.Survivin may inhibit the apoptosis of vascular endothelial cells in proliferative phase due to it's high expression.Caspase-3 and Surivivin involved in the hemangioma in the regulation of apoptosis.J Appl Clin Pediatr,2009,24(1):51-52

OBJECTIVETo mutate human annexin V gene and transform it to Pichia Pastoris for mutant human annexin V expression, so as to be purified as active annexin V with endogenous metal chelating site.

METHODSThe 5' and 3' end of native annexin V gene were mutated by specific primers. The mutant annexin V gene was inserted into pPIC9K and sequenced. The correct plasmid was linearized and transformed into Pichia Pastoris strain GS115 by electroporation. The transformants were selected from MD plates and cultured in BMGY medium and induced with methanol. The culture was centrifuged and the supernatant was analyzed by SDS-PAGE and silver staining. The binding activity of mutant human annexin V from culture supernatant was determined with phosphatidylserine exposed erythrocytes and fluorescein isothiocyanate-annexin V.

RESULTSThe 5' end of native human annexin V gene was fused with GCAGGCGGCTGCGGCCAT coding sequence and 3' end 946-948 site TGT was mutated to AGC. Pichia Pastoris transformants secreted proteins of relative molecular mass 36 000 48 h after methanol induction. The concentration of this protein that inhibited 50% of the binding of fluorescein-annexin V was 4nmol/L.

CONCLUSIONHighly-active recombinant mutant human annexin V with endogenous metal-chelating sites can be expressed in Pichia Pastoris system.

Annexin A5 ; genetics ; Base Sequence ; Humans ; Molecular Sequence Data ; Mutation ; Pichia ; genetics ; RNA, Messenger ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo evaluate the long term effects of adjuvant radiotherapy for postoperative breast cancer.

METHODSFrom 1985 to 1986, 162 patients with operable breast cancer were randomly given adjuvant radiotherapy according to clinical stage and involving condition of axillary lymph nodes (LN). The radiotherapy group (RG) was irradiated in the supraclavicular area and/or internal mammary area to 50 Gy, while the control group (CG) was not.

RESULTSThe overall 5-, 10- and 15-year survival rates of the RG were 72.0%, 56.1% and 54.3%, while they were 66.3%, 51.3% and 49.4% in the CG (P > 0.05). Clinical stage I-IIIa and positive or negative LN showed no significant difference in the two groups. But in patients with LN(+) > or = 4, the 5-, 10- and 15-year survival rates of the RG were 55.6%, 38.9% and 37.1%, which were higher than the CG of 29.0%, 16.1% and 16.1% (P < 0.05).

CONCLUSIONAdjuvant radiotherapy can improve the prognosis for breast cancer patients with LN(+) > or = 4, but not for LN(-).

Breast Neoplasms ; mortality ; radiotherapy ; surgery ; Female ; Humans ; Middle Aged ; Radiotherapy, Adjuvant ; Survival Rate

OBJECTIVETo study long-term behavioral and ultrastructural alterations in a hypoxic-ischemic brain damage (HIBD) model of neonatal rats.

METHODSSixty seven-day-old Sprague-Dawley rats were randomly subjected to unilateral carotid artery ligation followed by hypoxic exposure (HIBD group) or sham operation (n=30 each). A battery of behavioral tests, including Morris water maze test and sensorimotor tests, were performed at a postnatal age of 5 weeks. Nissl staining was used for counting neurons. Transmission electron microscopy was used for observing synapse structures and measuring the thickness of the postsynaptic density area and the length of the postsynaptic active area. The correlations of histological changes with the results of behavioral tests were evaluated.

RESULTSThe HIBD group showed a significantly longer escape latency (P<0.05) and a lower frequency of original platform crossing (P<0.05) in the Morris water maze test compared with the sham operation group. The sensorimotor function test showed that the sensorimotor function in the HIBD group was worse than in the sham operation group. Nissl staining showed that the number of neurons in the HIBD group was significantly reduced (P<0.01) compared with the sham operation group. Transmission electron microscopy showed that synapses were significantly reduced in number, and that the thickness of the postsynaptic density area and the length of the postsynaptic active area were reduced in the HIBD group. The thickness of the postsynaptic density area was negatively correlated with escape latency in the Morris water maze test (r=-0.861, P<0.01), and also negatively correlated with the total score of sensorimotor function tests (r=-0.758, P<0.05) in the HIBD group.

CONCLUSIONSHypoxia ischemia can lead to neuron loss and ultrastructure damage, resulting in long-term deficit of behavioral functions in neonatal rats.

Animals ; Animals, Newborn ; Brain ; pathology ; ultrastructure ; Female ; Hypoxia-Ischemia, Brain ; pathology ; psychology ; Male ; Maze Learning ; Microscopy, Electron, Transmission ; Rats ; Rats, Sprague-Dawley ; Reaction Time

BACKGROUNDMany studies on periostin have focused on its role in tumors and vascular reconstruction. However, the effect of periostin on stem cell function remains unclear. The aim of this study was to enhance vitality in adipose-derived stem cells (ADSCs), the effect of periostin on the function of ADSCs was observed.

METHODSHuman ADSCs (hADSCs) were isolated from human adipose tissue by collagenase I digestion and collected in multi-periods for in vitro culture. CD29, CD34, CD44, CD45 and CD105 were detected by flow cytometry. In addition, directed differentiation of hADSCs was induced using adipogenic, osteogenic and chondrogenic induction mediums. The induced morphological changes were observed using oil red O, Alizarin red and alcian blue staining. Periostin was administered to hADSCs in an acidic environment. The treatments of cells were divided into three groups: a periostin group (P); an acidic control group (A); a normal group (N). Then the resulting cell proliferation and migration were detected using a Cell Counting Kit-8 (CCK-8) and a transwell chamber assay, respectively.

RESULTSThe detection rates of CD29, CD44, CD105, CD34 and CD45 were 98.89%, 93.73%, 86.99%, 0.19% and 0.16%. The specific staining of cells was positive after induction culture. The mean absorbance of the cells in group P and A at 12 hours were 16.67% and 22.22% greater than group N, respectively (P < 0.01). The mean absorbance of cells from group P was 20.00% greater than that of group A at 48 hours (P < 0.05). The mean number of migratory cells per visual field in group A was 50.38% lower than that in group N (P < 0.05). The migratory cell number in group P was 119.98% greater than that in group A (P < 0.05).

CONCLUSIONSThe acidic environment impacted hADSC proliferation and inhibited cell migration. However, periostin was able to promote the proliferation and migration of hADSCs despite the acidic environment.

Adipose Tissue ; cytology ; Adult ; Antigens, Surface ; analysis ; Cell Adhesion Molecules ; pharmacology ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Humans ; Stem Cells ; drug effects ; physiology

AIM:To evaluate the effect of α2-macroglobulin(α2M) against X-ray induced obstacle on osteo-genic differentiation of human bone marrow mesenchymal stem cells(hBMMSCs). METHODS:hBMMSCs were cultured in vitro. The 4th generation of hBMMSCs was irradiated with 8 Gy X-ray,then induced osteogenic differentiation and trea-ted with different concentrations of α2M(0.5 and 1.0 g/L). The alkaline phosphatase(ALP) activity and the mRNA ex-pression of runt-related transcription factor-2 (RUNX2) were detected on day 7 after osteogenic induction. The protein ex-pression of osteoglycin (OGN) was evaluated by Western blot on day 14 after osteogenic induction. The formation of calci-um nodules was detected by alizarin red staining on day 21 after osteogenic induction. The activity of superoxide dismutase (SOD) and the protein expression of MnSOD of irradiated hBMMSCs with 8 Gy X-ray were determined at 24 h after α2M treatment. RESULTS:Compared with 8 Gy X-ray group,the activity of ALP,the mRNA expression of RUNX2,the pro-tein expression of OGN and MnSOD,as well as SOD activity were higher than those in the hBMMSCs treated with α2M at 0.5 and 1.0 g/L after 8 Gy X-ray irradiation,and the calcium nodules were also increased. CONCLUSION:α2M signifi-cantly improves the osteogenic differentiation ability,the SOD activity and MnSOD protein expression of hBMMSCs after ra-diation injury.