OBJECTIVETo observe the clinical effects of needle-pricking therapy, a newly medical and minimally invasive technique, for functional retrograde ejaculation and to explore its mechanism. Methods Thirty-six patients with functional retrograde ejaculation were randomly divided into an observation group(19 cases) and a control group(17 cases) In the observation group,needle-pricking therapy was used at Guanyuan(CV 4) and bilateral sacral plexus nerve,lumbar 1 nerve and greater occipital nerve stimulating points,once a week. In the control group, midodrine tablets were prescribed orally,three times a day. All the treatment was given for 9 weeks. The clinical effects of the two groups were observed, and the levels of luteinizing hormone(LH), testosterone(Tes) and estra4 diol(E2) were compared between the two groups.

RESULTSThe total effective rate of the observation group was, 89. 47%(17/19), which was better than 47.06% (8/17) of the control group(P<0. 05). The LH and Tes were obviously increased and E2 was decreased compared with those before treatment in the observation group(all P< 0. 01). Tes was raised(P<0. 05) and E2 was apparently declined in the control group(P<0. 01). After treatment, the differences of serum LH and Tes were statistically significant between the two groups(both P<0. 01).

CONCLUSIONNeedle-pricking therapy has advantages for functional retrograde ejaculation probably in that stimulating lumbosacral nerves can strengthen the function of pelvic floor muscles and urethral expansion muscle and regulate sexual gland axis.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Ejaculation ; Humans ; Male ; Middle Aged ; Needles ; Sexual Dysfunction, Physiological ; physiopathology ; therapy ; Treatment Outcome

Objective To investigate the dose- and time-effects of astragaloside Ⅳ(XGA) on collagen of myocardial fibroblasts in rats.Methods The myocardial fibroblasts of rats were separated by collagenase and trypsinase digestive method,and the cell culture system was established. After XGA in different concentrations and at different time points was administered in fibroblast culture systems,the mRNA expression levels of collagen,matrix metalloproteinases(MMP)-1,-2,-9,tissue inhibitor of metalloproteinase(TIMP)-1 and -2 were measured with reverse transcription-polymerase chain reaction(RT-PCR) test.Results After XGA administration with different doses and at different time points was adminstered,the gel electrophoresis product of RT-PCR in fibroblast culture system expressed the mRNA of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2;but the mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 decreased and the mRNA expression levels of MMP-1,-2,and -9 increased compared to those before XGA administration;the mRNA expression of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2 decreased or increased gradually with the increase of doses and the prolonged time of XGA use.The mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 were negatively related to the doses and action times of XGA(r=-0.927 to -0.637 P= 0 to 0.024);and the mRNA expression levels of MMP-1,-2 and -9 were positively related to the doses and action times of XGA(r=0.672 to 0.962 P=0 to 0.034).Conclusion XGA can markedly reduce the collagen formation of myocardial fibroblasts in rats,and its mechanisms are related to the inhibiting of collagen synthesis and the increase of collagen degradation.

Based on the Bacillus sp., isolated from Lop Nur Desert, the technology of separation and purification and the antioxidant effect were studied. After centrifugation and vacuum filtration, the deproteinization of supernatant was operated with Sevag reagent. The crude exopolysaccharide (EPS) was obtained by precipitation with ethanol. The optimum conditions for the isolation were as follow: pH 7.0, temperature 4?C, time 1.5 h, and material to ethanol ratio 1: 4. Dissolved in water, the crude EPS was fractional separated on activated carbon column (1.5 cm ? 24 cm), eluted with distilled water, 60% ethanol, 95% ethanol, and the main fraction was collected. Then the EPS was purified on Sephadex G-100 gel column, eluted with NaCl (0.2 mol/L). Fractions (4 mL, each) were also combined according to total sugar by phenol-sulfuric acid method and protein content was determined by Coomassie brilliant blue. The results showed that EPS was relatively homogeneous glycoprotein. The data of antioxidation in vitro showed that the EPS had a high antioxidant activity, which could quench hydroxyl radical, superoxide radical and had antilipid peroxidation activity. All of these indicated that EPS was a good natural antioxidant.

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

OBJECTIVETo investigate the effects of angiotensin converting enzyme inhibitor (ACEI) captopril on Calpain-mediated cardiomyocytes apoptosis and cardiac function in diabetic rats.

METHODSThirty adult male SD rats were randomly divided into 3 groups (n = 10), normal control group (NC group), diabetes mellitus group (DM group)and captopril treated group (Cap group). Streptozocin (STZ) were used to make the model of diabetes mellitus, captopril was administrated by gavage at the dose of 50 mg/kg every day, while in NC group and DM group the same volume of normal saline was administrated. Twelve weeks later, left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVDEP), maximal rise rate of left ventricular pressure (+ dp/dtmax) and maximal fall rate of left ventricular pressure (- dp/dtmax) were detected; Western blot was used to detect the expression of Calpain-1 Calpain-2, Bcl-2, Bax and total Caspase3 protein; apoptosis index (AI) were assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL).

RESULTSCompared with NC group, LVDEP was significantly higher; LVSP, + dp/dtmax and - dp/dtmax were significantly decreased (P < 0.05); Bcl-2 protein expression was decreased; the expression of Calpain-1, Calpain-2, Bax and total Caspase3 protein were increased; the value of AI was significantly increased. Compared with DM group, LVDEP was significantly lower; LVSP, + dp/dtmax and - dp/dtmax were significantly increased (P < 0.05); Bcl-2 protein expression was increased, the expression of Calpain-1, Calpain-2, Bax and total Caspase3 protein were decreased; the value of AI was significantly decreased (P < 0.05).

CONCLUSIONCaptopril can protect diabetic myocardial structure through inhibiting activation of Calpain-1 and Calpain-2, up-regulating the expression of Bcl-2, down-regulating the expression of Bax to inhibit Caspase3 dependent apoptosis, thereby improving the ventricular function and myocardial structure.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Apoptosis ; drug effects ; Calpain ; metabolism ; Cardiomyopathies ; pathology ; Caspase 3 ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; physiopathology ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Erythrocytes ; metabolism ; Fatty Liver ; blood ; Female ; Hepatitis ; blood ; Humans ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged ; Receptors, Complement 3b ; blood ; Thymosin ; therapeutic use ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.

METHODSForty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups: normal group, model group, control group (microwave), experimental group 1 (electricity), experimental group 2 (thermal), experimental group 3 (kneepad). Ten rabbits in the normal group were breed with conventional method; 9 rabbits in the model group were breed with conventional method after model made; 9 rabbits in the control group were treated with microwave for 30 minutes, one time daily; 9 rabbits in the experimental group 1 were treated with electricity (density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes, one time daily; 9 rabbits in the experiment group 3 were treated with electrothermal (OA knee pad) for 30 minutes, one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method.

RESULTSAt the 16 hthek,th e OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,wh ich indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,th e control group,th e experimental group 1 ,r oup 2,gr oup 3 were overexpressed,an d Belc2 mRNA expression levels of articular cartilage cells were low expression,an d compared with the normal group there were significant differences (P < 0.01). Belc2, p53 mRNA expression in articular cartilage cells,th ere were significant differences (P < 0.01) between the control group, experimental group 1, group 2, group 3 and model group. The results between the control group, experimental group 1 ,group 2 and group 3 had significant differences (P < 0.01).

CONCLUSIONOA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53, thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.

Animals ; Apoptosis ; physiology ; Disease Models, Animal ; Female ; Knee Joint ; metabolism ; pathology ; Male ; Osteoarthritis, Knee ; genetics ; pathology ; therapy ; Protective Devices ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics

Aneurysm ; complications ; therapy ; Cardiac Catheterization ; instrumentation ; Child ; Heart Septal Defects, Ventricular ; complications ; therapy ; Humans ; Septal Occluder Device

Objective To analyze the relation between ~(99)Tc~m-DTPA renal dynamic imaging and pathological changes in patients with lupus nephritis (LN).Methods Ten normal control and 29 patients with LN underwent ~(99)Tc~m-DTPA renal dynamic imaging.The LN patients were divided into two groups:silent LN (SLN) group,18 patients;and obvious LN (OLN) group,11 patients.For each case,glomerular filtration rate (GFR),peak time (t_p),half excretion time (t_(1/2)) and the excretion rate at 20 min (R_(20)) were calculated.Assessment of renal function on the scintigraphic images was evaluated by nuclear medicine physicians.The t-test,Fisher'exact probability and R×C association were used for data analysis.Results There were significant differences between normal people and two goups of LN in tp(t=5.3,9.3,both P＜0.05),t_(1/2)(t=6.9,12.0,both P＜0.05)and R_(20)(t=10.1,12.1,both P＜0.05).As to GFR,there was significant decrease in OLN patients(t=4.1,P＜0.05),but not in SLN patients(t=1.7,P＞0.05).Diagnoses of renal function by renal dynamic imaging were compared with the renal pathological changes (r=0.2273,P＜0.05).Conclusions ~(99)Tc~m-DTPA renal dynamic imaging is useful for evaluation of the early stage renal function for LN patients and to diagnose LN patients with no symptom of renal impairment.It may help to assess the degree of renal parenchymal damage while obviating the need for renal biopsy in these patients.

·AIM: To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell.· METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.· RESULTS: pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts.·CONCLUSIONS: Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.