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

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 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.

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

Objective To evaluate the performance of diluted thrombin time (dTT) assay for detecting Dabigatran levels and observe whether this assay may meet the requirements of clinical laboratory.Methods According to EP15-A2,EP6-A,EP7-A and C-24 documents of the Clinical and Laboratory Standards Institute (CLSI),the precision,trueness,analytical measurement range,carryover rate and anti-biological interference of dTT assay were evaluated and the stability of specimen for dTT assay was observed.Results Both the within-day and between-day coefficient of variation (CV) of dTT assay for detecting Dabigatran levels were consistent with manufacturer's stated CV.Compared with target values of Dabigatran,the relative bias of 3 levels of proficiency test materials from College of American Pathologists (CAP) were less than 10％.The results meet linear verification when Dabigatran concentration was between 30.92 and 249.13 ng/mL.The carryover rate was-0.84％.There was no interference for Dabigatran levels by dTT assay for detecting Dabigatran when Hb≤3 g/L,triglyceride≤873 mg/dL,heparin≤2.2 IU/mL and FDP≤29 mg/L.The results of stability showed that plasma specimens for dTT could not be stored at room temperature more than 4 hours,at 4 ℃ more than 4 days,at-20 ℃ exceed 1 month,while at-80℃ the plasma specimens could be stored at least 6 months for dTT assay.Conclusion The precision,trueness,analytical measurement range,carryover rate,anti-biological interference of dTT assay may meet the requirement of clinical laboratory.The stability of the specimen can fulfill the clinical requirements.

OBJECTIVETo compare the regulatory effects of electro-acupuncture (EA) at acupoints Zusanli (ST36) and Hegu (LI4) on the visceral hyper-sensitivity in the rat model of irritable bowel syndrome (IBS), and to explore the acting targets and specialty of acupoints.

METHODSExcept 8 rats of the normal control group, the rest 32 rats were prepared to set up the IBS models. IBS animal model was prepared by enema with acetic acid. Model rats were divided into three groups. Except for rats in the model group for control, those in the other two groups were treated 20 min by EA on ST36 (EA-ST36) and LI4 (EA-LI4) respectively for 2 weeks to observe the effect on behavior response of viscera sensitivity. The changes of neuropeptide (NPY), the somatostatin (SS) levels in blood and tissues of brain and intestine were monitored as well.

RESULTSThe volume thresholds for abdomen uplifting and back hunching were obviously increased after EA-ST36 (P<0.05), but showed insignificant change after EA-LI4. NPY contents lowered and SS contents increased in model rats; both EA-ST36 and EA-LI4 could raise the level of thalamic NPY (P<0.01 and P<0.05, respectively), but showed insignificant effects on NPY in colonic tissue. As for SS content, its colonic level could be reduced by EA-S36 and EA-LI4 (P<0.01 and P<0.05, respectively), however, its blood level was affected only by EA-ST36 (P<0.05).

CONCLUSIONSEA-ST36 or EA-LI4 could regulate the NPY in thalamus and SS in colonic tissue, the former could affect blood level of SS as well. It is deemed that NPY and SS may be the key substances for regulating the action of acupuncture in the brain-intestinal axis; their different levels could be regarded as an indicator for the functional difference between the acupoints.

Acupuncture Points ; Animals ; Brain ; metabolism ; Electroacupuncture ; methods ; Irritable Bowel Syndrome ; metabolism ; physiopathology ; Neuropeptide Y ; metabolism ; Rats ; Rats, Wistar ; Somatostatin ; metabolism ; Viscera ; physiopathology

OBJECTIVETo observe the effects of transcutaneous electrical acupoint stimulation (TEAS) combined with general anesthesia or controlled hypotension on hippocampal neuronal damage and the inflammatory response in peripheral circulation and central nervous system (CNS) after surgery, and to investigate its brain protection mechanism.

METHODSEighteen healthy male beagles aged 6 - 8 months were randomly divided into a general anesthesia group (group G), a controlled hypotension group (group C) and a compound anesthesia acupuncture group (group A), 6 cases in each group. Dogs in group G was anesthetized by isoflurane inhalation, and group C was combined with intravenous infusion of sodium nitroprusside based on isoflurane inhalation to induce hypotension, and followed surgery after achieving the target blood pressure, and group A was combined with TEAS at "Quchi" (LI 11), "Hegu" (LI 4) "Zu sanli" (ST 36) and "Sanyinjiao" (SP 6) based on controlled hypotension, and then brain tissue was taken out on the 72 h after mean arterial pressure (MAP) was returned to baseline levels. The concentration of IL-1beta,TNF-alpha in serum at different time points were detected by ELISA. The expression of IL-1beta, TNF-alpha, Bcl-2, Bax and cleaved caspase-3 were measured by immunohistochemistry, and the apoptosis of hippocampus were detected by TUNEL.

RESULTS(1) At different time points, the concentration of TNFalpha showed the trend of increase first and then decrease, while IL-1beta concentration represented a trend of decrease first and then increase in both group C and group A, but there were no significant differences in cytokine expression between the two groups (all P > 0.05). (2) The ratio of positive cells of IL-1beta, TNF-alpha and caspase-3 in CA1 and CA3 of hippocampus in both group C and A were higher than those in group G (all P < 0.01), and cytokines expression in group A were lower than those in group C (all P < 0.01), and caspase-3 in CA1 in group A was lower than that in group C (P < 0.01). The ratio of Bcl-2/Bax in both group C and A were lower than that in group G (all P < 0.01), and that in group A was higher than that in group C (P < 0.01 in CA1, P < 0.05 in CA3). (3) The apoptosis index (AI) of hippocampal neurons in both group C and A was significantly higher than that in group G (P < 0.01), while AI in CA1 in group A was lower than that in group C (P < 0.01).

CONCLUSIONThe TEAS can regulate the expression of inflammatory factor in hippocampus in animals undergoing general anesthesia or con trolled hypotension surgery, further improving Bcl-2/Bax ratio, inhibiting the expression of caspase-3 and reducing neuron apoptosis in hippocampus so as to play a neuroprotection.

Acupuncture Analgesia ; Acupuncture Points ; Anesthesia, General ; Animals ; Apoptosis ; Dogs ; Hippocampus ; cytology ; immunology ; surgery ; Humans ; Hypotension, Controlled ; Inflammation ; genetics ; immunology ; physiopathology ; therapy ; Interleukin-1beta ; genetics ; immunology ; Male ; Neurons ; cytology ; immunology ; Transcutaneous Electric Nerve Stimulation ; Tumor Necrosis Factor-alpha ; genetics ; immunology