OBJECTIVE: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. METHODS: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. RESULTS: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. CONCLUSION: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.
Apoptosis* ; Chromatin ; Chromomycin A3 ; DNA Nucleotidylexotransferase ; Fertility ; Humans ; In Situ Nick-End Labeling ; Male ; Semen ; Semen Analysis ; Spermatozoa* ; Sperm Head ; Protamines ; World Health Organization

Fatal anaphylactic reactions to protamine sulfate during cardiac surgery are very rare. We report a case of catastrophic bronchial spasm due to an anaphylactic reaction to protamine. The patient was managed successfully using a bronchodilator, steroid treatment, and extracorporeal membrane oxygenation.
Anaphylaxis* ; Bronchial Spasm* ; Extracorporeal Membrane Oxygenation ; Humans ; Protamines ; Thoracic Surgery

The detection of sperm DNA damage, as an important supplement to semen routine examination strategies, has been applied in some clinical andrology laboratories. What factors may lead to sperm DNA damage remains one of the concerns among many andrologists. Present studies show a variety of factors of sperm DNA damage, including age, environmental pollutants such as organophosphorus and organochloride pesticides, plasticizer, heavy metals such as lead, carcinogens such as polycyclic aromatic hydrocarbons (c-PAHs) and zearalenone (ZEA), male reproductive system diseases or systemic diseases such as varicocele, infection, tumor, spermatogenesis and maturation dysfunction, spinal cord injury and endocrine disorders, seasons and temperature, lifestyle, abstinence time, semen refrigeration, semen handling in vitro, and certain medications. Among them, spermatogenesis and sperm maturation dysfunction may be the most secretive factors, which are involved in the molecular mechanisms of sperm chromatin packaging and restructuring, such as the transformation of histone to protamine, single nucleotide polymorphism of genes, and the role of telomere, which may be one of the hotspots in the future studies of sperm DNA damage. Relevant researches in the future are expected to focus on the prevention of sperm DNA damage and clarification of its specific pathogenic mechanisms so as to provide some evidence for its treatment.
Age Factors ; Chromatin ; chemistry ; DNA Damage ; Environmental Pollutants ; toxicity ; Humans ; Male ; Protamines ; Semen ; drug effects ; Specimen Handling ; Spermatogenesis ; Spermatozoa ; drug effects ; Telomere ; physiology ; Varicocele ; complications

Protamine (PRM) is one of the most abundant arginine-rich nucleoproteins in sperm and plays an important role in spermatogenesis. In the late stage of spermatogenesis, the replacement of PRM by histone prompts the closer combination between the nuclear matrix of sperm and nucleoprotein in order for high enrichment and condensation of nuclear chromatin in addition to preventing the sperm genome from mutation induced by internal and external factors. With the development of DNA sequencing techniques, researches on the association between PRM polymorphisms and male fertility are surfacing as a hot field. Many studies show that rs2301365 polymorphism is a risk factor for male infertility and increases the risk of male infertility by 27 - 66%, that rs737008 polymorphism of PRM1 and rs1646022 polymorphism of PRM2 are protective factors against Asian infertility, and that the ratio of PRM1 to PRM2 is intensively associated with male infertility. This review presents an update on the association between PRM gene polymorphisms and male infertility.
Asian Continental Ancestry Group ; Humans ; Infertility, Male ; genetics ; Male ; Mutation ; Polymorphism, Single Nucleotide ; Protamines ; genetics ; Risk Factors ; Spermatogenesis ; Spermatozoa

OBJECTIVETo construct a single-chain human anti-EGFR antibody (scFv) and truncated protamine (tP) fusion protein, ScFv/tP, carrying small interfering (si)RNA directed against the heat shock protein Hsp47, a collagen-binding glycoprotein, in order to evaluate the role Hsp47 in apoptosis of hepatic stellate cells.

METHODSA single chain of the human variable fragment was obtained by phage display and fused with the tP gene and with or without (negative control) the Hsp47 siRNA sequences. Following expression and purification of the scFv/tP fusion protein and the scFv/tPHsp47 siRNA fusion protein, internalization capabilities were tested in isolated human hepatic stellate cells and the QSG-7701 human hepatocyte cells with visualization by immunofluorescent staining. The DNA binding ability of the fusion proteins were verified by gel shift assay.Following ScFv/tP-Hsp47 siRNA fusion protein transfection into the human hepatic stellate cells, the levels of Hsp47 mRNA and protein expression were tested by RT-PCR and Western blotting; in addition, effects of siRNA-mediated silencing of Hsp47 on cell proliferation and apoptosis were analyzed by the cell counting kit (CCK)-8, flow cytometry and Western blot detection of the apoptosis marker poly (ADP-ribose) polymerase (PARP).

RESULTSIndirect immunofluorescence revealed that the ScFv/tP fusion proteins were internalized into human hepatic stellate cells but not into the QSG-7701 cells.The ScFv/tP-Hsp47 siRNA fusion protein caused reduced expression of Hsp47 mRNA and protein expression in the human hepatic stellate cells, as well as increased the cells' apoptosis remarkably.

CONCLUSIONThe ScFv/tP fusion protein can be used as a transfection reagent to deliver Hsp47 siRNA into hepatic stellate cells and to mediate apoptosis via blockade of Hsp47 expression.

Apoptosis ; Cell Proliferation ; HSP47 Heat-Shock Proteins ; genetics ; Hepatic Stellate Cells ; cytology ; Humans ; Protamines ; metabolism ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Epidermal Growth Factor ; immunology ; Single-Chain Antibodies ; Transfection

OBJECTIVETo investigate the association of single nucleotide polymorphism (SNP) of the Protamine 1 (PRM1) gene in infertile men with teratozoospermia.

METHODSWe collected semen samples from 157 infertile men with teratozoospermia (case group) and 37 age-matched male volunteers (control group), and subjected them to morphological analysis. We extracted genome DNA, genotyped the polymorphism of the PRM1-190C- > A SNP (rs2301365) using the Sequenom MassARRAY system, compared the genotype frequencies between the case and control groups, and analyzed the sperm morphological parameters of different genotypes in the infertile males with teratozoospermia.

RESULTSThe frequencies of the genotypes CC, CA and AA were 38.9% (61), 44.6% (70) and 16.6% (26) in the case group, as compared with 45.9% (17), 51.4% (19) and 2.7% (1) in the control, with that of AA significantly higher in the patients than in the volunteers (P<0.05). The frequencies of the alleles C and A were 57.6% and 42.4% in the former, with no significant differences from 71.6% and 28.4% in the latter (P>0.05). Nor were any statistically significant differences observed in sperm morphology parameters between the genotype CC and CA, AA and CA + AA in the male patients (P>0.05).

CONCLUSIONThe SNP of PRM1-190C- > A might be associated with teratozoospermia-induced male infertility in the Han Chinese. Although this SNP may attribute to abnormal sperm morphology, the targeted part of sperm remains unclear.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Protamines ; genetics ; Spermatozoa ; abnormalities

PURPOSE: To determine the optimal combination of commercially available superparamagnetic iron oxide (SPIO) nanoparticles with transfection agents (TA). MATERIALS AND METHODS: Protamine sulfate (Pro) and poly-L-lysin (PLL) were incubated with ferumoxide and ferucarbotran in human mesenchymal stem cells at various concentrations, and cellular viability were evaluated. Cellular iron uptake was qualitatively and quantitatively evaluated. Cell visibility was assessed via MR imaging and the T2-relaxation time was calculated. RESULTS: The cellular viabilities with ferucarbotran were more significantly decreased than those with ferumoxide (p < 0.05). Iron uptake with ferumoxide was significantly higher than that for those with with ferucarbotran. The T2-relaxation time was observed to be shorter with ferumoxide in comparison to those with ferucarbotran (p < 0.05). Ferumoxide at a concentration of 25 microg/ml in combination with either Pro or PLL at a concentration of 3.0 microg/ml did not adversely impact cell viability, maximized iron uptake, and exhibited a lower T2-relaxation time in comparison to other combinations. CONCLUSION: Stem cells with ferumoxide exhibited a higher cellular viability and iron uptake in comparison to ferucarbotran- treated stem cells. A 25 microg/ml of ferumoxide with a 3.0 microg/ml of TA is sufficient to label mesenchymal stem cells.
Cell Survival ; Contrast Media ; Dextrans ; Ferric Compounds ; Humans ; Iron ; Magnetite Nanoparticles ; Mesenchymal Stromal Cells ; Nanoparticles ; Protamines ; Stem Cells ; Transfection

OBJECTIVETo establish a modified plasma protamine paracoagulation test.

METHODSPlasma protamine paracoagulation, modified plasma protamine paracoagulation and D-dimer (D-D) tests were performed for the plasma samples collected from 98 cases of disseminated intravascular coagulation (DIC) and 156 normal subjects. The sensitivity and specificity of the 3 tests were analyzed. The plasma samples from 8 cases of suspected myocardial infarction were detected using modified plasma protamine paracoagulation for diagnostic purpose.

RESULTSThe sensitivity of plasma protamine paracoagulation, modified plasma protamine paracoagulation and D-D tests was 16.33%, 88.76% and 77.56%, and the specificity was 100%, 88.46% and 97.44%, respectively. Positive results occurred earlier in modified plasma protamine paracoagulation test than in plasma protamine paracoagulation and D-D tests in 5 cases of myocardial infarction.

CONCLUSIONThe modified plasma protamine paracoagulation test has a higher sensitivity than plasma protamine paracoagulation test and a higher specificity than D-D test, and can be helpful in early diagnosis of thrombosis and fibrinolysis.

Adult ; Blood Coagulation Tests ; methods ; Female ; Humans ; Male ; Middle Aged ; Protamines ; blood ; Sensitivity and Specificity ; Thrombosis ; blood ; diagnosis

PURPOSE: Premixed insulin is effective to improve glycemic control; however, clinicians may be less likely to know which premixed insulin is appropriate for which patients. This study aimed to evaluate the effects of twice-daily injections of premixed insulin lispro on glycemic control in type 2 diabetic patients. MATERIALS AND METHODS: Forty type 2 diabetic patients, who had been treated with twice-daily injections of human protamine mixture 30/70 insulin for at least 12 months, were divided into two groups; one group whose blood glucose 2 hours after breakfast was greater than 200 mg/dL, was switched to lispro mix50, and the other group whose blood glucose 2 hours after breakfast < 200 was switched to lispro mix25. RESULTS: Glycated haemoglobin (HbA1c) significantly improved in the Mix50 group from 8.3% to 7.5% (at 12 weeks; p < 0.05), and to 7.5% (at 24 weeks; p < 0.05). On the other hand, HbA1c levels in the Mix25 group were slightly decreased from 8.1% to 7.7% at 12 weeks (p < 0.05), and to 7.9% at 24 weeks (not significant). Both postprandial plasma glucose and fasting plasma glucose levels were significantly improved in the Mix50 group, but not in the Mix25 group. Overall, 95% of subjects preferred premixed lispro insulin from human insulin in the viewpoint of the timing of insulin injection by questionnaire analysis. CONCLUSION: Switching from human protamine mixture 30/70 insulin to lispro mix50 twice-daily injection therapy in patients with high postprandial plasma glucose could improve their glycemic control and quality of life.
Aged ; Blood Glucose/*analysis ; Body Mass Index ; Body Weight ; Diabetes Mellitus, Type 2/*drug therapy ; Female ; Hemoglobin A, Glycosylated/metabolism ; Humans ; Insulin/administration & dosage/*analogs & derivatives ; Male ; Middle Aged ; Postprandial Period ; Protamines/administration & dosage ; Treatment Outcome

PURPOSE: Adenosine triphosphate (ATP) from the urothelium acts as a sensory neurotransmitter and is augmented in many diseases, such as overactive bladder. We investigated the effects of intravesical instillation of oxybutynin on ATP-induced bladder overactivity to determine whether this effect is mediated by effects on urothelial muscarinic receptors. MATERIALS AND METHODS: Cystometry (at rate of 0.04 ml/min) was performed in female Sprague-Dawley rats (body weight 250 g) under urethane anesthesia (1.2 g/kg). After a 2-hour baseline period, protamine sulfate (10 mg/ml) was instilled for 1 hour, and then ATP (60 mM, pH 6.0) or a mixture of oxybutynin (10(-6) M) and ATP (60 mM, pH 6.0) was instilled intravesically. We performed experiments with 4-diphenylacetoxy-N-methylpiperidine methobromide (4-DAMP) and methoctramine by the same methods. Cystometric parameters, such as the intercontraction interval (ICI), pressure threshold (PT), and maximal voiding pressure (MVP), were compared. RESULTS: With intravesical instillation of ATP after protamine sulfate treatment, the ICI was decreased compared with baseline (ICI: baseline, 487.1+/-64.8 s; protamine, 450.6+/-56.1 s; ATP, 229.7+/-35.3 s; p<0.05). Addition of oxybutynin, 4-DAMP, or methoctramine in the ATP solution did not significantly change the ICI compared with ATP solution alone (ICI: oxybutynin, 189.1+/-32.3 s; 4-DAMP, 161.1+/-22.8 s; methoctramine, 341.0+/-113.3 s; p>0.05). Intravesical instillation of ATP decreased MVP and PT significantly compared with baseline, but MVP and PT were not changed significantly with oxybutynin, 4-DAMP, or methoctramine compared with ATP. CONCLUSIONS: Bladder overactivity induced by intravesical instillation of ATP was not suppressed by intravesical instillation of antimuscarinics. Suppression of ATP-induced bladder overactivity by intravenous oxybutynin is not mediated by urothelial muscarinic receptors.
Adenosine ; Adenosine Triphosphate ; Administration, Intravesical ; Anesthesia ; Animals ; Diamines ; Female ; Humans ; Hydrogen-Ion Concentration ; Mandelic Acids ; Muscarinic Antagonists ; Neurotransmitter Agents ; Piperidines ; Polyphosphates ; Protamines ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; Urethane ; Urinary Bladder ; Urinary Bladder, Overactive ; Urothelium