Objective To observe dynamically osteopontin(OPN) expression in the renal tubules of streptozotocin(STZ) induced diabetic rats, and to explore the relationship between OPN and angiotensinⅡ(AngⅡ), nuclear transcription factor-?B(NF-?B), and renal injuries. Methods Male SD rats were injected with STZ to induce diabetes mellitus, which were randomly divided into 5 groups and meanwhile there were other 5 age-matched normal control groups. Immunohistochemistry was employed to observe the expression of OPN, AngⅡ, NF-?B and fibronectin(FN) in renal tubules. The OPN and I-?B protein in renal cortex was detected by Western blot methods. Blood glucose, serum creatinine and 24 h urine protein were examined. The renal morphology was checked through light microscopy.Results The AngⅡand NF-?B expression from DM day 3, OPN expression from day7, FN from week 2 in renal cortex or tubules was increased as compared with control groups, while I-?B protein in renal cortex was gradually decreased since day 3. In DM week 4, there were positive correlations between OPN and AngⅡ, NF-?B, FN and 24 h urine protein. Conclusion An increase of OPN expression in the renal tubules of diabetic rats may be regulated by AngⅡ and NF-?B, and therefore participates the injury mechanisms of renal tubulo-intestitium.

OBJECTIVETo explore the anti-aging effect of artemisia burning products (ie. smoke of moxibustion) and its proper intervention parameters.

METHODSAccording to factorial experiment design, 70 SAMP8 mice were randomly divided into one model group (group M) and 6 intervention groups: low concentration with 15 min group (group A1), low concentration with 30 min group (group A2), middle concentration with 15 min group (group B1), middle concentration with 30 min group (group B2), high concentration with 15 min group (group C1), high concentration with 30 min group (group C1). There were 10 cases in each group. Ten age-matched SAMR1 mice were used as normal group (group Z). All the mice in the 6 intervention groups were fumed with artemisia burning products of different concentration and time. The content of serum malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and glutathione reductase (GSH-Px) were tested.

RESULTSMDA content in group M was significantly higher than that in group Z (both P < 0.05), while SOD and GSH-Px activity were significantly lower in group M than that in group Z (both P < 0.05). Results of MDA, SOD and GSH-Px in 6 intervention groups were either of no statistically significant differences, or better than that in group M. Among 6 intervention groups, results of MDA and GSH-Px were better in group B1, while the result of SOD was better in group B2. Time factor didn't make any difference, while concentration of artemisia burning products is meaningful. As to SOD and GSH-Px, there's a strong interaction between the two factors.

CONCLUSIONWith certain concentration and time period, the intervention of artemisia burning products can exert anti-aging effect by increasing antioxydative capability and reducing metabolites of free radicals. Middle concentration and 30 minutes are recommended when intervened with artemisia burning products.

Aging ; blood ; metabolism ; Animals ; Antioxidants ; metabolism ; Artemisia ; chemistry ; Disease Models, Animal ; Glutathione Peroxidase ; blood ; Glutathione Reductase ; blood ; Humans ; Male ; Malondialdehyde ; blood ; Mice ; Moxibustion ; Superoxide Dismutase ; blood

OBJECTIVETo explore the mechanism of acupuncture for treatment of perimenopausal syndrome.

METHODSThe rats of perimenopausal syndrome were randomly divided into 3 groups, including an acupuncture group treated with acupuncture, a medication group with Gengnian'an, and a perimenopausal control group, with young rats used for a control group. Granulocyte apoptosis and expressions of Bcl-2 and Fas proteins in the ovary of the rat were detected.

RESULTSGranulocyte apoptosis increased significantly (P < 0.01), expression of Bl-2 proteins decreased significantly (P < 0.01) and expression of Fas proteins increased significantly (P < 0.01) in the ovary of perimenopausal rats as compared with the young rats; after acupuncture treatment, granulocyte apoptosis decreased significantly (P < 0.05), expression of Bel-2 proteins increased significantly (P < 0.05) and expression of Fas proteins decreased significantly (P < 0.01); after treatment of Gengnian'an, granulocyte apoptosis did not significantly change (P > 0.05), expression of Bcl-2 prteins increased significantly (P < 0.05) and expression of Fas proteins decreased significantly (P < 0.01).

CONCLUSIONAcupuncture can inhibit granulocyte apoptosis, up-regulate expression of Bcl-2 proteins and down-regulate expression of Fas proteins in the ovary of the perimenopausal rat.

Acupuncture Therapy ; Animals ; Apoptosis ; Female ; Granulocytes ; pathology ; Ovary ; metabolism ; pathology ; Perimenopause ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; analysis

Calcium sensitizers exert positive inotropic effects without increasing intracellular Ca(2+). Thus, they avoid the undesired effects of Ca(2+) overload such as arrhythmias and cell injury, but most of them may impair myocyte relaxation. However, MCI-154, also a calcium sensitizer, has no impairment to cardiomyocyte relaxation. To clarify the underlying mechanisms, we examined the effects of MCI-154 on Ca(2+) transient and cell contraction using ion imaging system, and its influence on L-type Ca(2+) current and Na(+)/ Ca(2+) exchange current with patch clamp technique in rat ventricular myocytes as well. The results showed that: (1) MCI-154 (1-100 micromol/L) had no effect on L-type Ca(2+) current; (2) MCI-154 concentration-dependently increased cell shortening from 5.00+/-1.6 microm of control to 6.2+/-1.6 microm at 1 micromol/L, 8.7+/-1.6 microm at 10 micromol/L and 14.0+/-1.4 microm at 100 micromol/L, respectively, with a slight increase in Ca(2+) transient amplitude and an abbreviation of Ca(2+) transient restore kinetics assessed by time to 50% restore (TR(50)) and time to 90% restore (TR(90)); (3) MCI-154 dose-dependently increased the electrogenic Na(+)/ Ca(2+) exchange current both in the inward and the outward directions in rat ventricular myocytes. These results indicate that MCI-154 exerted a positive inotropic action without impairing myocyte relaxation. The stimulation of inward Na(+)/ Ca(2+) exchange current may accelerate the Ca(2+) efflux, leading to abbreviations of TR(50) and TR(90) in rat myocytes. The findings suggest that the improvement by MCI-154 of myocyte relaxation is attributed to the forward mode of Na(+)/ Ca(2+) exchange.
Animals ; Calcium ; physiology ; Calcium Channels, L-Type ; drug effects ; Calcium Signaling ; drug effects ; Cardiotonic Agents ; pharmacology ; Cell Separation ; Cells, Cultured ; Dose-Response Relationship, Drug ; Heart Ventricles ; cytology ; Myocardial Contraction ; drug effects ; Myocytes, Cardiac ; cytology ; metabolism ; Patch-Clamp Techniques ; Pyridazines ; pharmacology ; Rats ; Rats, Wistar ; Sodium-Calcium Exchanger ; drug effects ; physiology

Objective To investigate the glucose tolerance and ability of insulin secretion in SD rats in second and third trimesters of pregnancy. Methods SD rats with pregnancy of 15 d were selected as experimental group(n=6),and another 6 rats of the same batch without pregnancy were served as controls(n=6).Intraperitoneal glucose tolerance trials(IPGTT) were conducted in these two groups.Rat islets were isolated after in situ collagenase digestion through pancreatic duct perfusion,islet morphology was observed by inverted phase contrast microscope,and insulin secretion was determined by radioimmunoassay. Results It was revealed by IPGTT that the levels of glucose at 30,60,90 and 120 min were significantly lower in experimental group than those in control group(P

Objective: To observe the therapeutic effect of mild moxibustion on irritable bowel syndrome (IBS) visceral hyperalgesiamodel rats and its regulatory effect on P2X3 receptors in the spinal cord, anterior cingutate cortex (ACC) and thalamic ventral posterolateral nucleus (VPL). Methods: Thirty 8-day-old newborn rats were randomly divided into a normal group (n=6) and a modeling group (n=24) according to the completely random number table method. Rats in the normal group were bred routinely, and those in the modeling group were subjected to preparing IBS chronic visceral hyperalgesia model using colorectal distention (CRD) in stimulation method. Rats successfully modelled were re-divided into a model group, a mild moxibustion group, a P2X3 receptor antagonist group, and a normal saline group according to the completely random number table method with 6 rats in each group. Rats in each group received corresponding interventions from the 37-day old, once a day for 7 consecutive days. Immunohistochemistry and Western blot assays were used to detect P2X3 protein expressions in the spinal cord, ACC and VPL of rats. Results: Under different intensities of CRD stimulation, the abdominal withdrawal reflex (AWR) scores of the model group were significantly increased versus the normal group (all P<0.05); the AWR scores of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group (all P<0.01). The P2X3 protein expressions in rat spinal cord, ACC and VPL tissues of the model group were significantly increased versus the normal group (all P<0.01); the P2X3 protein expressions in rat spinal cord, ACC and VPL tissues of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group (all P<0.01). Conclusion: Mild moxibustion can inhibit the P2X3 receptor expressions in the spinal cord, ACC, and VPL tissues of IBS visceral hyperalgesia model rats, which may be the mechanism of mild moxibustion in relieving the central sensitization of rats with IBS visceral hyperalgesia.

OBJECTIVETo establish a rapid and simple method to detect Y chromosome microdeletions directly using whole blood as starting material for multiplex-PCR.

METHODSUsing a self-prepared pHpH-Bufferq, multiplex-PCR amplification was directly carried out from the anticoagulant whole blood sample without DNA extraction step. Twelve sequence tagged sites (STS), namely SY84, SY86, SY127, SY134, SY124, SY132, SY152, SY157, SY239, SY242, SY254 and SY255, in AZFa, AZFb, and AZFc gene regions were detected in 5 different tubes. In order to ensure the validity of the experiments, sex-determining region Y (SRY) and X-linked or Y-linked zinc finger gene (ZFX/Y) were used as internal controls. Furthermore, conventional PCR using genomic DNA extracted from each blood sample was performed in parallel for evaluating the accuracy of the experiments.

RESULTSA total of 156 male samples were detected, and a normal male sample and a female sample were used as a positive and a negative control respectively. The results showed that 144 samples had no deletion; one sample was AZF-deleted; one sample was AZFb-deleted; seven samples were AZFc-deleted; one sample was both AZFb- and AZFc- deleted; and two samples were all AZFa-, AZFb- and AZFc- deleted. The observed results from two kinds of starting material (whole blood and purified DNA) are completely consistent.

CONCLUSIONIn our method, PCR amplification was directly carried out from whole blood without any DNA extraction step. So it has the advantages of low cost, simple process, time-saving operation and less cross-contamination. The whole process can be completed within 2 hours. Thus the efficiency of clinical detection is improved greatly.

Azoospermia ; genetics ; Cells, Cultured ; Chromosome Deletion ; Chromosomes, Human, Y ; Female ; Humans ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Sex Chromosome Aberrations

The purpose of this study was to compare the detection of trisomy 8 in myelodysplastic syndrome (MDS) patients with interphase fluorescence in situ hybridization (FISH) and cytogenetic karyotype analysis. Using Spectrum Green labeled chromosome 8 centromere probe, interphase FISH was established. The trisomy 8 clones were simultaneously detected in 48 MDS cases with FISH and conventional cytogenetic analysis (CCA). Results showed that the CCA revealed no significant difference of constitutional proportion between MDS-RA and MDS-RAEB with karyotypes of whole +8, partial +8 and one +8. With FISH, detectable rates were 66.1% for whole +8. Partial +8 and sole +8 were significantly higher than one +8 and complex +8, respectively. The percentages of trisomy 8 were similar in MDS-RA and MDS-RAEB. Trisomy 8 was detected in 1 of 15 specimens with normal or abnormal karyotype without trisomy 8 by FISH. There was linear correlation between the percentages of partial +8 detected by FISH and CCA. Two patients received CCA and FISH examination at diagnosis and during treatment, the percentage of trisomy 8 was increased with progress of disease. In conclusion, our results showed that FISH is a sensitive and accurate technique to detect trisomy 8 in MDS patients. It can provide contribute to diagnosis, assessment of curative effect and predicting progress of disease in MDS. Clone size of trisomy 8 does not related to classification of MDS, but sole +8 is seems to see in MDS-RA frequently.
Chromosomes, Human, Pair 8 ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Myelodysplastic Syndromes ; genetics ; pathology ; Reproducibility of Results ; Sensitivity and Specificity ; Trisomy

OBJECTIVETo evaluate the protective effect of high dose ambroxol, a mucoactive drug, on acute lung injury caused by paraquat in rats.

METHODSOne hundred and thirty-six healthy male Sprague-Dawley rats were randomly divided into three groups: control group (n = 24) injected with normal saline intraperitoneally, PQ group (n = 56) [(2% paraquat (25 mg/kg) injected into peritoneal cavity on the first day)] and AT group (n = 56) ambroxol 35 mg/kg was injected into peritoneum daily after paraquat intoxication once daily for 7 consecutive days. The arterial gas was determined and the extent of lung injury was assessed by measuring the ratio of wet to dry weight (W/D) and protein content in BALF, the WBC count, the percentage of PMN, the content of malondialdehyde (MDA) and the levels of superoxide dismutase (SOD) in the blood and BALF respectively. Left lung tissue was observed through both light microscope and electron microscope (TEM).

RESULTSThe white cell count and the content of protein in the blood and the BALF of PQ group were significantly higher than those of the control group (P < 0.05 or P < 0.01). On the 7th day, the content of MDA 9 [(8.12 +/- 1.12) nmol/ml] in the serum of PQ group was significantly higher than the control group and the GSH-Px activity [(1256.8 +/- 133.2) U/ml] was significantly lower than the control group (P < 0.01). The white cell count and the content of protein in the blood and the BALF of AT group were significantly lower than the PQ group (P < 0.05 or P < 0.01). On the 7th day, the content of MDA in the serum of the AT group [(4.86 +/- 0.75) nmol/ml] was significantly lower than the PQ group and the GSH-Px activity [(1509.5 +/- 183.0) U/ml] and the SOD activity [(3903.2 +/- 374.7) U/ml] were significantly higher than the PQ group (P < 0.01). Under optical and electronic microscopes, the injury of lung tissue was reduced after large dose of ambroxol was administered.

CONCLUSIONTreatment with ambroxol (35 mg/kg) could influence the status of oxidative stress in lung and alleviate lung injury induced by paraquat. Ambroxol has obviously therapeutic effect on paraquat poisoning.

Acute Lung Injury ; chemically induced ; drug therapy ; metabolism ; pathology ; Ambroxol ; pharmacology ; therapeutic use ; Animals ; Disease Models, Animal ; Lung ; drug effects ; metabolism ; pathology ; Male ; Oxidative Stress ; Paraquat ; poisoning ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of ambroxol on paraquat poisoning induced acute lung tissue injury and the change of pulmonary surfactant associated protein A in the experimental rats.

METHODSOne hundred and twenty healthy adult male Sprague-Dawley rats were randomizedly assigned into normal saline (NS) group (n = 24), paraquat poisoning induced lung tissue injury model (PQ) group (n = 48) and ambroxol treatment (AT) group (n = 48). The indexes were observed among the three groups comprising the mortality rate, the change of arterial blood PaCO(2) and PaO(2), the ratio of wet to dry lung tissue (W/D), the change of the lung tissue under light and electric microscope respectively, and the expression of pulmonary surfactant associated protein A.

RESULTSThe mortality rate of rats in the PQ group was 50.0% on the seventh day while the mortality rate in the AT group was 25.0%. The level of arterial blood PaCO(2) in the PQ group (6.94 +/- 0.8) kPa was significantly higher than that in the AT group (6.12 +/- 0.5) kPa and the NS group (4.6 +/- 0.4) kPa. The level of arterial blood PaO(2) in the PQ group (6.98 +/- 1.1) kPa was significantly lower than that in the AT group (8.25 +/- 0.7) kPa and the NS group (12.7 +/- 0.8) kPa. There were significant differences among the groups (P < 0.05). The degree of lung tissue injury was severe in PQ group and relieved in AT group. The expression of pulmonary surfactant associated protein A was significantly decreased in PQ group 13.22% +/- 2.21% on the seventh day, compared with that in the AT group (21.82% +/- 3.67%) (P < 0.05). The expression of pulmonary surfactant associated protein A in AT group was significantly higher in the AT group (18.97% +/- 0.91%) than that in the PQ group on the seventh day (P < 0.05).

CONCLUSIONAmbroxol plays a role in facilitating synthesis and secretion of pulmonary surfactant protein A and relieves the lung tissue injury induced by paraquat poisoning.

Ambroxol ; pharmacology ; Animals ; Immunohistochemistry ; Lung ; metabolism ; pathology ; Male ; Paraquat ; poisoning ; Pulmonary Surfactant-Associated Protein A ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; pathology