BACKGROUND: As a biochemical marker of cytoplasm, the increased activity of creatine kinase (CK) in human spermatozoa is correlated with both the residual cytoplasma and the ratio of sperm with abnormal func tion. It is a marker of mature sperm and associated with the potential of in semination. OBJECTIVE: To investigate the differences of CK activity in sperm and the relative contents of sperm CK-MM and CK-BB isoenzymes between normal fertile males and infertile males, and evaluate its significance in clinical diagnosis of male infertility. DESIGN: Case-control observation. SETTING: Laboratory of Biochemistry, Family Planning Research Insti tute of Sichuan Province. PARTICIPANTS: Ninety-four male infertile patients between January 1999 and October 2000 were selected from the Department of Family Planning Sciences of Sichuan Province, who had no aspermatism with their wives proved to be fertile. The average age of subjects were 31 years. Eighty subjects with the sperm count ＞ 2×1010 L-1 were taken as normal sperm group and 14 subjects with the sperm counts ＜ 2×1010 L-1 were considered as oligospermia group. Semen obtained from 27 healthy males who were normal in routine examinations and with children was taken as the healthy control group.METHODS: Semen sample collected by masturbation after abstinence of 3 to 5 days was incubated at 37 ℃ for liquefication and routinely analyzed.Total activity of CK in sperm was determined by using a kinetic spectrophotometry and the relative contents of CK isozyme was determined by agarose gel electrophoresis followed by density scanning of CK isozyme.MAIN OUTCOME MEASURES: Sperm counts, percentages of viability and motility of sperm, total CK activity and the relative contents of CK-MM and CK-BB isozyme in spermatozoa.RESULTS: A total of 94 enrolled patients and 27 normal controls were involved in the analysis of results. ①Sperm counts, percentage of viability and motility in oligospermia group ( Ⅱ + Ⅲ, WHO method) were obviously lower than those in the healthy control group, and those in the normal sperm group, except the sperm counts, were remarkably lower than the healthy control group [the sperm counts in healthy control group, normal sperm group and oligospermia group was (6.05 ±0.81 )×1010 L-1, (7.76±1.37)×1010 L-1 and (1.46±0.19)×1010 L-1 respectively (P ＜ 0.01). The survival rate in healthy control group, normal sperm group and oligospermia group was 85.1%,56.8% and 58.2%, P ＜ 0.01, and the sperm motility was 62.9% ,34.6% and 29.5% respectively, P ＜ 0.01].②Total activity of sperm CK in oligospermia group was significantly higher than the healthy control group [respectively (9.000±6.117) and (1.933 ±0.943) kat/108 sperm,P＜ 0.05],although that in the normal sperm group (2.800±0.862) kat/108 sperm was a little higher than the healthy control group, while there were no significant difference between the two groups (P ＞ 0.05).③The relative contents of sperm CK-MM isozyme in the normal sperm group and oligospermia group were obviously decreased (that in the healthy control group, normal sperm group and oligospermia group was 30.5%, 19.0% and 18.0% respectively,P ＜ 0.05), which implied a remarkable difference in sperm differentiation between healthy control group and the latter two groups.CONCLUSION: The determination of sperm CK is meaningful for the diagnosis and pathogenesis of oligospermia. More work should be done on the distribution of sperm CK-MM isozyme in different infertile population as well as its importance in the diagnosis of infertility.

Objectives　To investigate the relationship between homocysteine (Hcy), folic acid, vitamin B12(VitB12),lipids and the carotid artery atherosclerosis(CAAs) and the significance of their levels in predicting cardiac and cerebral vascular events of older patients. Methods　Both sides of the common carotid artery and internal carotid artery were measured by B-mode ultrasound in 74 examinees in A, B, C, and D groups and a control group. With fasting serum, cholesterol, triglyceriade, apolipoprotein AI(Apo AI), apolipoprotein B (Apo B), lipoprotein (a) ［Lp(a)］, creatinine, glucose, folic acid, Vit B12 and Hcy were detected. Results　When carotid artery atherosclerosis became severer, serum Hcy was higher. The levels of the control and A, B, C, D groups were (10.2±3.6) μmol/L, (11.5±4.4) μmol/L, (17.9±4.5) μmol/L, (24.7±10.3) μmol/L, and (41.4±22.3) μmol/L respectively. SNK test showed a significant difference between D and other groups (P>0.05). Folic acid and Vit B12 were negatively correlated with the severity of carotid artery atherosclerosis, but only A and D groups showed statistical significance. Spearman correlation coefficient showed that serum Hcy had significant negative correlation with folic acid and Apo AI(P=0.035 and P=0.000).Multiple linear regression analysis between CAAs and various risk factors showed Apo AI had negative correlation, but Apo B, Lp(a) and Hcy had positive correlation with CAAs, especially Hcy(P=0.009).Conclusion　Hyperhomocysteinaemia is an important risk factor of artery atherosclerosis and the degree of CAAs is highly correlated with Hcy concentration. Hcy, Apo AⅠ,Apo B, Lp(a) are sensitive and reliable indexes for evaluating and predicting CAAs.

Objective To establish a PCR-RFLP method for detection of CD26 mutation of hemoglobin E(HbE)gene.Methods The genome DNA from the family members who were suspected of having HbE combined with beta-thalassemia was amplified using PCR.The PCR products were digested by restriction endonuclease Mnl I,and then separated on PAGE.The electrophoretical patterns were finally analyzed to confirm if CD26 mutation was present or not.Results Four cases with CD26 mutation in two families were successfully screened out.Conclusion Although the incident frequency of CD26 mutation in Sichuan population is not high,it must not be neglected since it may present alone or combined with other type of thalassemia.PCR-RFLP method described in this study is available in screening mutated HbE gene and treating the disease owing to its simplicity,rapidity and specificity.

Objective To report the clinical and myopathological features in a patient with common variable immunodeficiency (CVID) with myositis.Methods A 33 years old man suffered from recurrent respiratory infection with fever over 10 years.The symptoms improved after anti-infection therapy.At the same time he presented with fatigue.Two years ago he developed general muscle weakness,hypertrophy and myotonia,especially in the hands,neck and thighs.Genetic test for myotonic dystrophy protein kinase (DMPK) and zinc finger protein 9 (ZNF9) was performed.Laboratory tests,electromyography,muscle ultrasound and muscle biopsy were performed.In addition to standard histological and enzyme histochemical stainings,immunohistochemical method was used with primary antibodies of mouse anti human monoclonal antibodies including CD8 for T-lymphocytes,CD20 for B-lymphocytes,CD68 for macrophages and MHC-Ⅰ for muscle membrane.Results Electromyography revealed myogenic changes and abound with myotonic potentials.There was muscle hypertrophy in muscle ultrasound.Lung biopsy showed chronic inflammatory changes.Serum hypoimmunoglobulin and anemia were found.Muscle biopsy showed muscle fiber necrosis and regeneration with lymphocyte and macrophage infiltration.There were no gene mutations in DMPK and ZNF9 gene.Conclusion Muscle hypertrophy and myotonia appeared in CVID with myositis.

OBJECTIVETo explore the effects of arecoline on hepatic insulin resistance in type 2 diabetes rats and to elucidate its possible mechanism.

METHODSForty five Wistar rats were fed with high fructose diet for 12 weeks to induce type 2 diabetic rat model. rats were randomly divided into 5 groups (n = 8): control group, model group and model group were treated with different dose (0, 0.5, 1, 5 mg/kg) of arecoline. After 4 weeks, the fasting blood glucose, blood lipid and insulin level measured , mRNA expression of liver constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by reverse transcription polymerase chain reaction (RT-PCR), the protein expression of p-AKT and glucose transporter4 (GLUT4) were detected by Western blot.

RESULTS1.5 mg/kg arecoline could significantly decrease the level of fasting blood glucose, blood lipid, blood insulin level and liver G6Pase, PEPCK, IL-6, TNF-alpha mRNA level in type 2 diabetes rats. 1.5 mg/kg arecoline also could significantly increase CAR, PXR mRNA level and p-AKT and GLUT4 protein expression.

CONCLUSIONArecoline improved hepatic insulin resistance in type 2 diabetes rats by increasing the mRNA levels of CAR and PXR leading to the creased glucose metabolism and inflammation related genes expression.

Animals ; Arecoline ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Glucose-6-Phosphatase ; metabolism ; Insulin Resistance ; Interleukin-6 ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Phosphoenolpyruvate Carboxykinase (GTP) ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Receptors, Steroid ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Fluidized-bed manufactured enteric-coated diclofenac sodium pellets were compressed into tablets. The blend of two aqueous acrylic resins dispersion in different ratios, Eudragit NE30D and Eudragit L30D-55, were used to prepare enteric-coated diclofenac sodium pellets of different particle sizes and coating level. The cushioning pellets with different properties and these enteric-coated pellets were compressed into tablets in different proportions. The drug release of the tablets containing these pellets would be lower than 10% in 2 h in simulated gastric fluid, but reach (83 +/- 2.42)% in 1 h in simulated enteric fluid. The mixture of Eudragit NE30D and Eudragit L30D-55 could be used to prepare enteric pellets which are suitable for compression. The cushioning pellets which were composed of stearic acid/microcrystalline cellulose (4:1, w/w) could avoid rupture of the coating of pellets during the compression.
Acrylic Resins ; chemistry ; Anti-Inflammatory Agents, Non-Steroidal ; chemical synthesis ; Cellulose ; chemistry ; Diclofenac ; chemical synthesis ; Drug Carriers ; Drug Compounding ; methods ; Drug Delivery Systems ; Methacrylates ; chemistry ; Particle Size ; Polymers ; chemistry ; Solubility ; Tablets, Enteric-Coated ; chemistry

A new mathematical equation characterizing the compression of pharmaceutical materials is presented. This equation presumed that the rate of change of the compressible volume of powder with respect to the pressure is proportional to the compressible volume. The new model provided a good fit to several model substances employing non-linear regression techniques. The validity of the model had been verified with experimental results of various pharmaceutical powders according to the Akaikes informatics criterion (AIC) and the sum of squared deviations (SS). The parameter of the new model might reflect quantitatively the fundamental compression behaviors of the powders. It had demonstrated that the proposed model could well predict the compaction characteristics of solid particles like the Kawakita model.
Compressive Strength ; Nonlinear Dynamics ; Powders ; chemistry ; Pressure

Microcrystalline cellulose (MCC), calcium phosphate (DCP)/MCC (4:1, w/w) and lactose (Lac)/MCC (4:1) pellets with different intragranular porosity were prepared in an extrusion-spheronizator and three volume ratios of ethanol/water were used as binder agents to prepare pellets. The compression behaviors of these pellets with different intragranular pore volume were evaluated with the parameters of Kawakita model. The results showed that high pore volume of pellets made up of MCC had the best compressibility and low pore volume of pellets had a poor compactibility. However, the compressibility of different porosity of pellets made up of DCP/MCC (4:1) or Lac/MCC (4:1) was good, but they were not significantly different. The reason might be the main compression mechanism of high porosity of MCC pellets was plastic deformation and that of DCP/MCC pellets or Lac/MCC pellets was not plastic deformation but fragmentation. These results can be observed directly by the SEM photographs. According to these results, the conclusion could be drawn that high porosity MCC pellets and different porosity DCP/MCC pellets and Lac/MCC pellets can be used as cushion granules to maintain the original shape and release characteristics of drug pellets when pellets were tabletted.
Calcium Phosphates ; chemistry ; Cellulose ; chemistry ; Drug Compounding ; methods ; Excipients ; Lactose ; chemistry ; Microspheres ; Porosity ; Pressure ; Tablets

Objective To observe the effect of inducible costimulator(ICOS) costimulation pathway blockade in rat limb allografts acute rejection by RNA interference. Methods Twenty-seven cases of modified hind llmb allotransplantation were performed from Wistar to SD rats. The rats were divided into 3 gronps(each n=9): the rejection group not given a special disposal; the control group, consisting of SD rats that received injection of pSilencer 4.1 and Sofast complex by vein post transplantation; and the interference group that received injection of pSilencer 4.1-ICOSshRNA and Sofast complex. On the eighth day posttransplantation, 3 rats were killed to study the pathological changes in each group. The expressions of ICOS gene in vivo were detected by flow cytometry and RT-PCR. The mixed lymphocyte reaction (MLR) was performed and eytokines in blood were measured by ELISA. The rest rats were used to record limb survival time. Results The mean survival time in rats of the rejection and the control groups were(11.34±1.21) and (11.14±1.32) days respectively. In the interference group, the mean survival time of limb allografts was (16.85±1.73) days(P<0.05). The rats in the rejection and the control groups experienced moderate to serious acute rejections with skin epidermal necrosis, a large quantity of lymphocyte infdtration, muscle cell necrosis and interstitial edema, while the pathological changes in rats of the interference group were mild. The splenocyte ICOS mRNA expression level in the interference group(18.75%) was significantly lower than that of the rejection group(100%) and the control group(98.51%). ICOS cell surface expression level as judged by the fluorescence intensity was 45.59±12.87 in the interference group, 103.72±21.76 in the rejection group, and 93.47±29.55 in the control group(F=6.89, P<0.05). In stimulation assays, a one-way mixed lymphocyte reaction stimulation index(SI), with spleen cells from Wistar and Lewis rats, respectively, the rejection group (5.26±0.42,5.18±0.29) and the control group (5.37±0.27,4.93±0.44) had significantly greater reactions than the interference group(2.37±0.35, 4.87±0.36), respectivily(F=7.29, P<0.05; F=6.19, P0.05). In the IFN-γ and IL-4 expression assays, reactions of the interference group (230.17±38.47,160.32±59.13) were lower than those of the rejection group(490.73±51.48,230.67±45.21) and the control group(480.15±43.96, 240.53± 47.36), (F=7.23,6.75, all P<0.01). Conclusions In vivo transfection of pSilencer 4.1-ICOS shRNA interference plasmid can effectively block T-cell co-stimulation pathway, suppress acute rejection, and prolong limb allografts survival.

Leaf senescence is often caused by water deficit and the chimeric gene P(SAG12)-IPT is an auto-regulated gene delaying leaf senescence. Using in vitro leaf discs culture system, the changes of contents of chlorophylls, carotenoids, soluble protein and thiobarbituric acid reactive substance (TBARS) and antioxidant enzymes activities were investigated during leaf senescence of P(SAGl2)-IPT modified gerbera induced by osmotic stress compared with the control plant (wild type). Leaf discs were incubated in 20%, 40% (w/v) polyethylene glycol (PEG) 6000 nutrient solution for 20 h under continuous light [130 micromol/(m(2) x s)]. The results showed that the contents of chlorophylls, carotenoids and soluble protein were decreased by osmotic stress with the decrease being more pronounced at 40% PEG, but that, at the same PEG concentration the decrease in the transgenic plants was significantly lower than that in the control plant. The activities of superoxide dismutase (SOD), catalases (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and dehydroascorbate reductase (DHAR) were stimulated by PEG treatment. However, the increases were higher in P(SAG12)-IPT transgenic plants than in the control plants, particularly at 40% PEG treatment. Lipid peroxidation (TBARS content) was increased by PEG treatment with the increase being much lower in transgenic plant than in the control plant. It could be concluded that the increases in the activities of antioxidant enzymes including SOD, CAT, APX, GPX and DHAR were responsible for the delay of leaf senescence induced by osmotic stress.
Alkyl and Aryl Transferases ; genetics ; Antioxidants ; metabolism ; Arabidopsis Proteins ; genetics ; Ascorbate Peroxidases ; Asteraceae ; genetics ; metabolism ; Carotenoids ; metabolism ; Catalase ; metabolism ; Chlorophyll ; metabolism ; Cysteine Endopeptidases ; genetics ; Genes, Bacterial ; Genes, Plant ; Lipid Peroxidation ; Osmotic Pressure ; Oxidoreductases ; metabolism ; Peroxidase ; metabolism ; Peroxidases ; metabolism ; Plant Leaves ; metabolism ; Plant Proteins ; metabolism ; Plants, Genetically Modified ; Promoter Regions, Genetic ; Solubility ; Superoxide Dismutase ; metabolism