Child ; Diagnosis, Differential ; Humans ; Male ; Rhabdomyosarcoma, Embryonal ; diagnosis ; therapy

OBJECTIVETo investigate the role of lead in the expression of the renal fibrosis related nuclear factor kappaB (NF-kappaB), transforming growth factor (TGF-beta) and fibronectin (FN) in rat kidney and the possible molecule mechanism of lead induced renal fibrosis.

METHODSThirty-two Sprague-Dawley rats were randomized into 4 groups. Group A was fed with distilled water as control group. Group B, C and D were fed with the water including 0.5% lead acetate continuously for 1, 2 or 3 months respectively. At the end of treatment, the expressions of renal NF-kappaB, TGF-beta and FN were detected by immunohistochemistry and RT-PCR.

RESULTSThe immunohistochemistry analysis showed that expressions of NF-kappaB in group B, C and D (0.2315 +/- 0.0624, 0.3213 +/- 0.0740, 0.4729 +/- 0.0839 respectively) were continuously increased as compared with that in group A (0.1464 +/- 0.0624). The RT-PCR analysis showed that expressions of NF-kappaB in group B, C and D (0.4370 +/- 0.0841, 0.5465 +/- 0.0503, 0.6443 +/- 0.0538 respectively) were also increased as compared with that in group A (0.3608 +/- 0.0550). However, there was no change for TGF-beta in 4 groups except that it was increased markedly in group D (0.5225 +/- 0.0416) as compared with that in group A (0.4645 +/- 0.0461) by RT-PCR. The expressions of FN in group C and D (0.4243 +/- 0.0595 and 0.4917 +/- 0.0891 by immunohistochemistry; 0.8650 +/- 0.0880 and 0.8714 +/- 0.0980 by RT-PCR) were increased as compared with those in group A (0.3530 +/- 0.0490 by immunohistochemistry and 0.7432 +/- 0.0639 by RT-PCR).

CONCLUSIONThe lead can increase the expression of renal NF-kappaB, TGF-beta and FN in rats, which may be related to the lead induced renal fibrosis in rats.

Animals ; Fibronectins ; biosynthesis ; genetics ; Fibrosis ; chemically induced ; Immunohistochemistry ; Kidney ; drug effects ; metabolism ; pathology ; Lead ; toxicity ; Male ; NF-kappa B ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; biosynthesis ; genetics

OBJECTIVETo investigate the effect of potassium iodide on the expression of nuclear factor-kappaB and fibronectin.

METHODSThe experiment was performed with 72 SD rats weighing about 180-220 g. The animals were randomly assigned into nine groups. Group A, B, C (n=8) served as control and were fed with distilled water for 1 month, 2 month, 3 month respectively. Group D, E, F (n=8) served as lead exposed and were fed with water with 0.5% lead acetate for 1 month, 2 month, 3 month respectively. Group G, H, I (n=8) served as potassium iodide and lead exposed and were treated with 0.5% lead acetate simultaneously taking potassium iodide 3 mg/100 g weight by intragastric administration for 1 month, 2 month, 3 month respectively. Animals of different groups were sacrificed at the end of the treatment. Ultrastructure of kidney was observed by electron microscopy; Expression of NF-kappaB and FN protein and mRNA in kidney were measured respectively by immunohistochemistry and RT-PCR.

RESULTSElectron microscopic examination revealed potassium iodide could restrain the denaturalization in epithelial cells and mitochondrial cristae. The expressions of NF-kappaB protein (0.2315 +/- 0.0624, 0.3213 +/- 0.0740, 0.4729 +/- 0.0839) and mRNA (0.4370 +/- 0.0841, 0.5465 +/- 0.0503, 0.6443 +/- 0.0538) in all the lead exposed groups continuously increased compared with correspondent control groups; Group I was decreased obviously compared with group F. The expressions of FN protein (0.4243 +/- 0.0595, 0.4917 +/- 0.0891) and mRNA (0.8650 +/- 0.0880, 0.8714 +/- 0.0980) in group E and F increased compared with group B and C, but the expressions of FN protein in group I significantly decreased compared with group F; The expressions of FN mRNA in Group H and I significantly decreased compared with group E and F.

CONCLUSIONThe potassium iodide can ameliorate renal ultrastructure and degrade expression of nuclear factor-kappaB and fibronectin induced by lead.

Animals ; Disease Models, Animal ; Fibronectins ; genetics ; metabolism ; Kidney ; drug effects ; metabolism ; ultrastructure ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Lead Poisoning ; complications ; metabolism ; pathology ; Male ; NF-kappa B ; genetics ; metabolism ; Potassium Iodide ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system. METHODS: The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/microL, separately. All samples were then amplified in 10, 5, 3 microL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci. RESULTS: Under the situation of 28 cycles and 3 microL volume, samples which achieved > 40 pg/microL could be correctly typed. Under the situation of 10, 5, 3 microL volume, samples which achieved > 20 pg/microL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band. CONCLUSION DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.
DNA/genetics* ; DNA Fingerprinting/methods* ; Feasibility Studies ; Forensic Genetics/methods* ; Humans ; Limit of Detection ; Male ; Polymerase Chain Reaction/methods* ; Semen/chemistry* ; Sensitivity and Specificity ; Tandem Repeat Sequences

BACKGROUNDOveractive bladder (OAB) can be caused by many factors such as inflammation, bladder outlet obstruction, neurogenic factors. We performed an intraperitoneal (ip) injection of cyclophosphamide to induce cystitis in rats, which causes their detrusors to overact, to provide a valuable disease model for discussing OAB pathogenesis and to study effective curing methods.

METHODSFemale Sprague-Dawley rats were induced to form cystitis by cyclophosphamide (200 mg/kg, ip). The day after the injection, two catheters were inserted into each rat's bladder to study its urodynamics. The BL-410 model bio-function experimental system was used to monitor bladder pressure while the rats were conscious. Unstable detrusor contractions appear in the urine storage period as a standard to determine OAB, and the positive rate was calculated. Urodynamic parameters such as bladder basal pressure (BP), maximum voiding pressure (MVP), intercontraction interval (ICI), spontaneous activity (SA), maximum cystometric capacity (MCC), and bladder compliance (BC) were recorded in each group, and a light microscope was used to observe the pathological changes in the rat bladder tissue.

RESULTSThe detrusor instability rate of the model group was 83.33%. The MVP, MCC and BC of rats in the model group were lower than the control group (P < 0.01), and the BP, ICI and SA of the model group rats were higher than the control group (P < 0.01). The difference between the control group and the model group is statistically significant. The model group rats' bladder walls swelled and bled, the submucosa thickened and leukocyte infiltration became serious.

CONCLUSIONSAcute cystitis and OAB symptoms can be induced by ip injections of cyclophosphamide in rats. This can provide a valuable animal model to study OAB in human beings.

Animals ; Consciousness ; Cyclophosphamide ; toxicity ; Female ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder, Overactive ; chemically induced ; physiopathology ; Urodynamics ; physiology

Objective:To observe the effect of blood stasis syndrome on renal damage and endoplasmic reticulum stress (ERS) in diabetic kidney disease (DKD) rats, and to explore the relationship between renal syndrome of blood stasis damage and ERS in DKD rats. Method:The 50 Male SD rats of SPF level were selected to establish DKD rat model by high fat and high sugar diet combined with intra-abdominal injection of streptozotocin(STZ). They were randomly divided into normal group, diabetes mellitus group and diabetes mellitus and blood stasis syndrome group（0.05 mg·kg^-1）, among which diabetes mellitus and blood stasis syndrome group was prepared by dextran method. The general conditions, hemorheology indexes, 24 h urine protein, serum creatinine and renal pathology of the rats in each group were observed. Immunohistochemical analysis, Western blot and Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were used to detect mRNA and protein expressions of endoplasmic reticulum stress-related proteins glucose regulated protein 78(GRP78), activating transcription factor-6(ATF6) and renal fibrosis index fibronectin(FN), and alpha smooth muscle actin(α-SMA). Result:Compared with normal group, the rats in diabetes mellitus group showed polyphagia, polyuria and weight loss, increased hemorheology index of whole blood viscosity and plasma viscosity(P<0.05，P<0.01), increased 24 h urine protein, serum creatinine and urea nitrogen(P<0.01), and increased renal pathology, and increased mRNA and protein expression of GRP78, ATF6, FN and α-SMA(P<0.05，P<0.01). After dextran preparation of blood stasis model. Diabetes mellitus and blood stasis syndrome group increased mortality, signs of change is more obvious in the diabetic group, whole blood viscosity, plasma viscosity, 24 h urine protein ration, serum creatinine and urea nitrogen were significantly higher than those in diabetic rats(P<0.01), pathological changes aggravated in the diabetes group. At the same time， mRNA and protein expressions of GRP78, ATF6, FN, and α-SMA in renal tissue were significantly higher than those in diabetic mellitus group(P<0.05，P<0.01). Conclusion:Under the combined disease and syndrome model, the blood stasis syndrome may further aggravate the pathological damage of the kidney of DKD rats, and is related to the enhancement of ERS in the kidney of DKD rats.