Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.

Objective To study the effects of YinlingⅠon expelling lead and the improvement of the ability of learning memory in lead poisoned mice.Methods Poisoning model of lead was prepared by drinking water with lead acetate,and the administration of YinlingⅠor EDTA-Na_2Ca to lead poisoned mice was performed.Lead content was detected in blood, brain and bone.The ability of lear- ning and memory of mice was measured monthly by Y-water maze test. Ultrastructure of CA3 cell in hippocampus was observed with transmission electron microscope.Results After administration of YinlingⅠ,the lead content in blood, brain and bone decreased remarkably, the ability of learning and memory increased,and the ultrastructure changes of CA3 cell in hippocampus markedly dimi- nished.Conclutions YinlingⅠ may expel lead of the mice with lead poisoning and improve their ability of learning and memory.

Objective To evaluate the efficacy and safety of acute ureteroscopy for the treatment of ureterie stones during middle and late pregnancy.Methods From June 1998 to March 2005,17 pregnant women(mean age,27 years;age range,21-35 years)with ureteric stones were treated by ureteroscopy when the fetus was at 20-36 weeks of gestation(mean,29 weeks).All the cases presented with urgent symptoms such as recurrent renal colic(11 cases),fever(4)or acute obstructive anuria(2).Among 17 cases,the stones(between 6 mm?7 mm and 13 mm?21 mm)were located in the upper(8 cases),middle(5)or lower ureter(4);and on the left side(5 eases),on the right(10)and on both(2)of the lower ureter. Mild hydronephrosis were observed in 6 cases and moderate hydronephrosis in 11,Of the 17 cases,14 under- went ureteroscopic pneumatic lithotripsy;in 1 case the calculi were pushed to the renal pelvis;and 2 cases were treated by Double-J catheter drainage.Results All the urgent symptoms in 17 cases were relieved after treatment.The stone-free rate of initial treatment was 82.4%(14 of 17).Three cases with residual stones were treated by Douhle-J catheters,which were replaced every 3 months until the calculi were re- moved.No abortion,premature delivery or complications such as ureter perforation occurred.Mild renal colic occurred in 1 case after insertion of Douhle-J catheter,and it was relieved 3d later;gross hematuria occurred in l case and disappeared 6 d later without treatment.All 17 patients had normal delivery and gave birth to healthy children.Conclusions Ureteroscopy is a safe and reliable method for the treatment of ureteric calculi during middle and late pregnancy.

BACKGROUNDHypoxia-inducible factor (HIF) may play an important role in the process of tumorigenesis as well as tumor progression. The aim of this study was to compare the expression between HIF-1α and HIF-2α in tumor angiogenesis and the overall impact on patient prognosis in human non-small cell lung cancer (NSCLC).

METHODSIn the current work we compared the immunohistochemical expression of HIF-1α and HIF-2α in surgical specimens of 140 patients with NSCLC in a tissue microarray study. Relationships between HIF-α expression and clinicopathological or angiogenic factors, including prognosis, were analyzed.

RESULTSHigh HIF-1α and HIF-2α expression was noted in 49/140 (35.0%) and in 64/140 (45.7%) of the cases, respectively. There was no direct correlation between HIF-1α and HIF-2α expression. Patients with advanced stage tumors had frequent high expression of HIF-2α (P = 0.007), and we also found a significant correlation between HIF-2α and T or N stage (P = 0.030 and 0.043, respectively). HIF-1α showed a marginal association with T stage (P = 0.084), which showed a higher expression in early stage tumors. A significant correlation (P = 0.045) was noticed between HIF-1α and vascular endothelial growth factor (VEGF) expression while the expression levels of thymidine phosphorylase (TP), cyclooxygenase (COX)-2 and microvessel density (MVD) were significantly higher in high HIF-2α tumors (P = 0.020, 0.004, and 0.046, respectively). In addition, univariate analysis of overall survival demonstrated that HIF-2α expression, but not HIF-1α, was related to poor outcome (P = 0.001) and it retained significant in multivariate analysis (P = 0.036).

CONCLUSIONSTaken together, we conclude that HIF-1α and HIF-2α may differentially regulate the major angiogenic factors in different stages of the tumor process in NSCLC. HIF-2α may play a dominant role in tumor angiogenesis and appears to be of obvious value as a significant prognostic factor in NSCLC.

Adult ; Aged ; Angiogenesis Inducing Agents ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Immunohistochemistry ; Male ; Microvessels ; Middle Aged ; Multivariate Analysis ; Prognosis ; Thymidine Phosphorylase ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Objective To explore achieving the consistent method of blood lipid examination by comparing the results of 5 dif-ferent blood lipid detection system commonly used in the use of refernce method to assign freach blood serum before and af-ter calibration.Methods Used the indoor quality control total variation (CV%)to evaluate the 5 blood lipid examination system of the imprecision.Referenced the United States Clinical and Laboratory Standardization Institution (CLSI)9A2 EP program,compared with 54 fresh blood serum in 5 commonly used examination system of Total Cholesterol (TC)and Tri-glyceride (TG),and then estimated the bias between the different detection systems and mean value.8 of the samples were determined by the reference method and estimate the bias of different system.The fresh frozen serum samples assigned by reference method were used to evaluate the above examination system,then compare and estimate the bias again with the same 54 fresh serum samples.Compared the variation of 54 samples in different detection system before and after calibra-tion.Results The TG imprecision of 5 examination system were between 3.76%~23.65%,the TC imprecision between 2.19%~23.43%,that mean the results were good,the r value of TG were between 0.996 7~0.999 6 and the TC were 0.956 2~0.996 7.But there were obvious differences between the results of the systems,and the biggest difference were 14.72%~34.21% in TG and 3.11%~14.57% in TC.After use the serum assignment by reference method,the variation of the systems has been significantly decreased.Conclusion Using the reference method to assign the fresh serum of different blood lipid detection system can effectively improve the consistency of the results.

OBJECTIVEPostoperative pain is a common problem in male circumcision. We investigated the effect of nanometer silver dressing (Shenzhen AGT Pharm. Co. Ltd.) in relieving pain following male circumcision.

METHODSSixty patients undergoing circumcision in the outpatient department were randomized into an experimental and a control group, the incision covered with nanometer silver dressing in the former and with vaseline dressing in the latter. None of the patients received any analgesics or other pain-killing therapies after surgery. The postoperative pain intensity was accessed using the modified numeric pain intensity assessment scale at 1, 2, 3, 5 and 7 days after the operation, and statistical analyses were performed using SPSS 12.0 software.

RESULTSThe patients averaged (31.13 +/- 13.94) years in age, and had no significant differences in age and body mass index (BMI) between the two groups. At 1, 2, 3 and 5 days, postoperative pain intensity was significantly lower in the experimental than in the control group (P > 0.05). Multiple regression analysis revealed that postoperative pain score was not correlated with patients' age, BMI and types of disease, but with the types of dressing.

CONCLUSIONNanometer silver dressing can significantly alleviate postoperative pain of circumcision, and is particularly applicable to such moist parts as the perineum, genitals, and urethra.

Adolescent ; Adult ; Bandages ; Circumcision, Male ; Humans ; Male ; Middle Aged ; Nanostructures ; therapeutic use ; Pain Measurement ; Pain, Postoperative ; therapy ; Silver ; pharmacology ; therapeutic use ; Treatment Outcome ; Young Adult

To investigate the feasibility of using Real-Time PCR to evaluate the effectiveness of Sindbis virus inactivation by Methylene Blue with visible light. Sindbis virus was treated by Methylene Blue with different intensity of visible light and the transcribed cDNA was quantified by Real-Time PCR. Residual infectivity of treated virus was tested by cell infection method as parallel control at the same time. The residual infectivity of virus decreased from 6.50 lgTCID50/mL to under the limit of detection as light intensity increased. Meanwhile, the quantity of virus cDNA decreased significantly (P < 0.05), which correlated to the decline of virus infectivity (R2 > 0.98). Methylene Blue with visible light could cause lesion to nucleic acid of Sindbis virus, the extent of which was light intensity-dependent and correlated to the decrease of virus infectivity. The results demonstrated that Real-Time PCR can be a useful tool for evaluating effect of virus inactivation after Methylene Blue treatment with light.
Light ; Methylene Blue ; pharmacology ; Polymerase Chain Reaction ; methods ; Sindbis Virus ; drug effects ; genetics ; physiology ; radiation effects ; Virus Inactivation ; drug effects ; radiation effects

Objective:To report the sampling study design and radiography protocol of a large-sample investigation on skeletal maturation of 3 to 18-year-old children in China.Methods:Multi-stage stratified random sampling was employed in this study. Two provinces, municipalities, or autonomous regions were randomly selected from each of the seven regions of China, including Northeast China, Northwest China, North China, Central China, East China, Southwest China, and South China. Then one rural and one urban investigation site were randomly selected from each province, municipality, or autonomous region. In total 28 sites were included. Among those sites, four residential districts were randomly selected from each urban site, and four townships from each rural site. For each residential district or township, 1-4 kindergartens, primary schools, and middle schools were chosen. Random cluster sampling was used to extract 3-<6-year-old children in kindergartens, and 6-18-year-old children in primary schools and middle schools. The investigation on skeletal maturation was sampled proportionate to the sampling of the whole study. The estimated simple size was 780 for each site, and 21 840 for all 28 sites in total. There were six groups of 3-<6-year-old children classified at 0.5-year intervals, and 12 groups of 6-18-year-old children classified at 1-year intervals. Posteroanterior position radiography of the left hand and wrist was achieved for all subjects.Results:The study was performed from August 26, 2019 to October 16, 2021. In total, 20 444 children received posteroanterior position radiography of the left hand and wrist, including 10 196 males and 10 248 females, 9 711 urban and 10 733 rural, respectively. The 3-<6-year-old group included 1 611 (male 819, female 792) subjects, and the 6 to 18-year-old group included 18 833 (male 9 377, female 9 456) subjects.Conclusion:This nationwide investigation on skeletal maturation of 3 to 18-year-old children in seven regions of China was successfully preformed. The results of this study can provide an important reference for establishing the current evaluation criteria of bone age in Chinese children and adolescents.

This study was aimed to investigate the influence of TLR2 and TLR4 agonists on the migration and adhesion activity of umbilical cord blood (UCB) CD34(+) cells and to explore the underlying mechanism. The expression of TLR2 and TLR4 on UCB CD34(+) cells was detected with flow cytometry. The effect of TLR2 agonist (PAM3CSK4) and TLR2 agonist (LPS) on the migration and adhesion ability of UCB CD34(+) cells was evaluated with chemotaxis and adhesion assays. The results indicated that expression levels of TLR2 and TLR4 were (14.2 ± 3.8)%, (19.6 ± 4.1)% respectively. Compared with the control group, the migration activity of UCB CD34(+) cells toward SDF-1 decreased significantly in LPS group (p < 0.01). The adhesion activity was not altered significantly in LPS group. However, both the migration activity towards SDF-1 and the adhesion activity of UCB CD34(+) cells were not changed significantly in PAM3CSK4 group. Further study found that LPS did not affect the expression level of CXCR4 on CD34(+) cells, but could inhibit the spontaneous migration ability of CD34(+) cells. It is concluded that TLR4 activation can decrease the chemotaxis function of CD34(+) cells towards SDF-1, which may associate with the decreased spontaneous migration ability of CD34(+) cells.
Antigens, CD34 ; blood ; Cell Movement ; drug effects ; Cells, Cultured ; Chemokine CXCL12 ; Fetal Blood ; cytology ; immunology ; Humans ; Lipopeptides ; pharmacology ; Lipopolysaccharides ; pharmacology ; Toll-Like Receptor 2 ; agonists ; Toll-Like Receptor 4 ; agonists

OBJECTIVEAttended to apply the acellular urinary bladder submucosa in the operation of the urethral duct reconstruction of penile hypospadias.

METHODSIn 2 cases, the urethral ducts were reconstructed by using the acellular urinary bladder submucosa as dorsal half of the urethra, while the preputial island flap as ventral side. In the other patient, the urethral plate was incised, similar to Snodgrass operation, the matrix was fixed to the corpora cavernosa beneath.

RESULTSUroflowmetry and the urethroscopy were been performed in the one year follow up. A bell-shaped flow curve was obtained in the patient after urethroplasty using matrix graft combined with a local flap, and no fistula, stricture was found after a year or half a year follow-up.

CONCLUSIONSThe freeze-dried acellular urinary bladder submucosa had three-dimensional network structure, and the acellular urinary bladder submucosa graft combined with preputial flap for hypospadias repair provided satisfactory functional results.

Child, Preschool ; Freeze Drying ; Humans ; Hypospadias ; surgery ; Male ; Mucous Membrane ; transplantation ; Transplantation, Homologous ; Urinary Bladder ; transplantation