Three-dimensional structure model of azoreductase AZR of Rhodobacter sphaeroides was con- structed using homology modeling method. It is a flavodoxin adopting ?/? structure. Structure alignment of two different types of flavin-dependent azoreductases revealed that they possessed high similarity. Based on sequence and structure analysis, site-directed mutagenesis of K109H and K109A were performed. The opti- mal pH values are pH 6 and pH 9 for K109H and K109A mutant protein, respectively. The optimal tempera- ture (30℃) is not affected by mutagenesis. Positively charged residues at position 109 is necessary for the binding of methyl red, while K109H is not a conserved mutagenesis for the binding of NADPH. K109 may only be involved in the binding of the 2’-phosphate group of NADPH and have no effect on the binding of NADH.

Objective To discuss methods of decoration reconstruction for finger defect in emergency and to observe the elinical effects.Methods Of the 41 cases of finger injuries of different degrees,15 were repaired with part of the skin flaps of the big toenails or skin flaps of the second toenalis,8 were repaired with part of the skin flaps of the big toenails,7 were reeonstructed with the second tiptoes,11 were repaired with the abdominal skin flaps of the big toes or lateral flaps of the second toes.Results All the 41 fingers sur- vived.One skin flap of the big toe was somewhat swelling and a decorating operation was performed.The 4～18 months of follow-up visitation of the rest cases revealed good function and shapes.No obvious functional ab- norality was found in the donating feet.Conclusion Various kinds of decoration reeonstruetion for finger defects are available to recover the hand shape and function as much as possible.

OBJECTIVETo observe the intervention effect of Leihong Granule (LG) in in-stent restenosis (ISR) after endovascular therapy for lower extremity arterial occlusive diseases (LEAOD).

METHODSRecruited 80 LEAOD patients who successfully underwent endovascular therapy (balloon dilation and stent implantation) were randomly assigned to two groups, the control group and the LG group, 40 in each group. Patients in the control group received basic treatment, while those in the LG group additionally took LG for 3 months. Plasma levels of IL-10, IL-18, CRP, and the intima-media thickness (IMT) of lower extremity artery were observed in the two groups between and after treatment. The rate of stent patency, ABI, intermittent claudication, rest pain, and the incidence of amputation the two groups were recorded and observed in the two groups.

RESULTSIn the control group, serum levels of IL-10, IL-18, CRP, and IMT were significantly higher one month after surgery than before surgery (P < 0.05). There was no significant difference in serum levels of IL-10, IL-18, CRP, or IMT between the two groups before surgery (P > 0.05). These indices were obviously lower in the LG group than in the control group after surgery (P < 0.05). Compared with the control group, the incidence rates of intermittent claudication and the rest pain at 6 months and 12 months after surgery significantly decreased (P < 0.05). The stent patency rate at 6 months and 12 months after surgery, and ABI were significantly higher than those of the control group (P < 0.05). There was no statistical difference in the amputation rate between the two groups (P > 0.05).

CONCLUSIONLG might effectively improve ischemic symptoms of affected limbs possibly through lowering the ISR rate after endovascular therapy for LEAOD through preventing immunosuppressive actions.

Aged ; Aged, 80 and over ; Arterial Occlusive Diseases ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Graft Occlusion, Vascular ; therapy ; Humans ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Phytotherapy ; Stents ; Treatment Outcome

OBJECTIVETraditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.

METHODSSix virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.

RESULTSThe sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.

CONCLUSIONThe method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.

Escherichia coli Infections ; diagnosis ; microbiology ; Escherichia coli Proteins ; genetics ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Shiga-Toxigenic Escherichia coli ; genetics ; isolation & purification ; Virulence Factors ; genetics

AIM: This study was designed to explore the effects of short-term and long-term pretreatment of diammonium glycyrrhizinate (GLN) on the pharmacokinetics of entecavir (ETV) in rats. METHODS: Male SD rats were randomized into short-term and long-term experimental groups, respectively. In the short-term experiment, the control group received saline, the low dose group received GLN 13.5 mg·kg(-1) and the high dose group received GLN 40.5 mg·kg(-1). ETV (0.09 mg·kg(-1)) was given i.g. 0.5 h after saline/GLN administration. For the long-term experiment, rats were allocated into two experimental designs. The control group received saline/ETV (0.09 mg·kg(-1)), the low dose group received GLN 13.5 mg·kg(-1)/ETV 0.09 mg·kg(-1) + GLN 13.5 mg·kg(-1), while the high dose group received GLN 40.5 mg·kg(-1)/ETV 0.09 mg·kg(-1) + GLN 40.5 mg·kg(-1); all administration was continued for 15 days. On the 16(th) day, 0.09 mg·kg(-1) ETV was administrated to all groups. Blood samples were obtained at different time points after ETV administration to determine plasma ETV concentrations. RESULTS: Pretreatment with glycyrrhizin resulted in no significant alterations in the main pharmacokinetic parameters of ETV in the short-term and long-term administration experiments. CONCLUSION Diammonium glycyrrhizinate has no effect on ETV pharmacokinetics in rats.
Animals ; Drug Interactions ; Glycyrrhizic Acid ; pharmacology ; Guanine ; analogs & derivatives ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To explore the application of quantitative temperature testing (QTT) in forensic identification and clinical diagnosis of neurogenic erectile dysfunction (NED). METHODS: TSA-II-NeuroSensory Analyzer was used to measure the thresholds of four kinds of sensory, including cold, cold pain, heat, heat pain, in 22 normal and 35 NED patients at dorsal glans (DG), left thigh interior (LTI) and left thenar (LT). To calculate the relative thresholds of the sensory mentioned above between DG and LTI (DG/LTI), and between DG and LT (DG/LT). Then to analyze those thresholds and the relative thresholds. RESULTS: NED group showed significant higher threshold than the normal group in DG-heat, DG-heat pain, LTI-heat, LTI-heat pain, DG/LTI-heat, DG/LT-heat, DG/LT-heat pain (P < 0.05). CONCLUSION The threshold of QTT at dorsal glans could be used as an accessory indicator in forensic medicine and clinical diagnosis of NED.
Adult ; Case-Control Studies ; Erectile Dysfunction/physiopathology* ; Hand/physiology* ; Humans ; Male ; Nervous System Diseases/physiopathology* ; Neurologic Examination/methods* ; Pain Threshold ; Penis/physiopathology* ; Sensory Thresholds ; Temperature ; Thermosensing

OBJECTIVEThere are a great number of modified models based on the four-vessel occlusion (4VO) model of Pulsinelli and Brierley which has been used worldwide for brain ischemia research. However, up to now the problems of collateral circulations of 4VO and the difficulty in arranging a surgery to occlude the basilar artery in other models are not satisfactorily solved yet. In this study, an improved 4-vessel occlusion (I4VO) rat model which is easy to handle and able to decrease the effect of collateral circulation is reported.

METHODSThe common carotid arteries and the beginning of the subclavical arteries of rats were occluded for different time by one-stage apertura thoracis superior approach. Neurological deficit scores defined by the modified Garcia scoring system and histopathological method were used to evaluate the effects of this model up to 7 days after reperfusion.

RESULTSThe neurological scores in the 15-min and 25-min groups decreased significantly at 24, 48 and 72 hours after reperfusion (P less than 0.05), and the histopathologic study showed that there were stable, symmetrical changes of lesions in bilateral hippocampus in all the ischemia samples from two ischemia groups compared with sham operated group (P less than 0.05).

CONCLUSIONThis modified model is safe, easy, reliable, stable, mini-invasive as well as time-saving in making bilateral hemispheric ischemia, which can effectively decrease collateral circulations and meanwhile lead to stable lesions in hippocampus and cortex.

Animals ; Brain Ischemia ; Hippocampus ; Rats ; Rats, Sprague-Dawley

The present study was to investigate the effects of intracerebroventricular (i.c.v.) injection of interleukin-1beta (IL-1beta) on thermal nociception in SD rats. The rats were divided into control and drug-administration groups. The animals of control group were given vehicle solution via i.c.v. injection. The animals of drug-administered groups were given IL-1beta at different doses (5, 50 and 500 pg/kg b.w.) via i.c.v. injection. IL-1 receptor antagonist (IL-1ra, 50 ng/kg) was injected 20 min before injection of IL-1beta or vehicle solution. The nociceptive threshold, which was represented as paw withdrawal latency (PWL), to a noxious thermal stimulation was measured using an analgesiameter. I.c.v. injection of IL-1beta dose-dependently shortened the PWL. At the dose of 500 pg/kg, the shortening of the PWL occurred at 20 min, reaching a peak within 40 min, lasted 100 min after injection, then gradually returned to the baseline level. Pretreatment with IL-1ra completely blocked the effects of IL-1beta-induced shortening in PWL. The results obtained suggest that IL-1beta may induce hyperalgesia in rats through binding to IL-1 receptors in the brain.
Animals ; Hot Temperature ; Hyperalgesia ; drug therapy ; Injections, Intraventricular ; Interleukin 1 Receptor Antagonist Protein ; pharmacology ; Interleukin-1beta ; pharmacology ; Nociception ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin-1 ; antagonists & inhibitors ; metabolism ; Touch

Objective To analyse the preliminary clinical results of intensity modulated radiation therapy (IMRT) for 122 untreated nasopharyngeal carcinoma (NPC)pafients.Methods 122 NPC pa- tients received IMRT alone from Feb.2001 to Jun.2004,with 31 females and 91 males,and a median age of 45 years(range 25-66).According to the Fuzhou Stage Classification,there were StageⅠ11 patients, StageⅡ34,StageⅢ62,and StageⅣa 15.IMRT was carried out using an inverse planning system (COR- VUS 5.0,Peacock plan) developed by the NOMOS Corp.The treatment was given with the Multi-leaf Inten- sity Modulating Collimator (MIMIC) using a slice-by-slice arc rotation approach.The prescription dose was 68 Gy/30f to the nasopharynx gross tumor volume (GTV_(nx)),60-66 Gy/30f to positive neck lymph nodes (GTV_(nd)),60 Gy/30f to the first clinical target volume (CTV_1) and 54 Gy/30f to the second clinical target volume (CTV_2).Kaplan-Meier method was used to calculate the overall survival rate (OS),distant metas- tasis-free survival rates (DMFS),and local-regional control rates from the last date of therapy.Log-rank test was used to detect the difference between groups.Results The median follow-up time was 20 months ( range 6 to46 months).The 1-,2-,and 3-year OS was 95.2%,91.4%,85.1%,DMFS was 91.9%, 88.6%,85.6%,and the local-regional control rates was 96.5%,93.2%,93.2%,respectively.Statistics of the local control rate was insignificant either for advanced T(T3+T4) stage or early T(T1+T2) stage diseases(P=0.148).The 2-year regional control rate was insignificant either for patients with N(+) or N (-),but the 2-year DMFS was significant both for patients with N(+) and N(-)lesions(P=0.004).For 17 patients who failed,there were two with residual disease and one with recurrence at the primary site (17.6%),three patients in the neck (17.6%),twelve patients (70.6%) in distant metastases.Conclu- sions Intensity modulated radiation therapy does provide excellent local-regional control for untreated NPC, especially in patients with advanced T stage or N(+) lesion.Distant metastasis is the main cause of failure. N (+) is significantly correlated with distant metastasis.

Brachytherapy is a key part of radical radiotherapy for cervical cancer. Along with the persistent development of radiotherapy techniques, the combined application of brachytherapy and intensity-modulated radiotherapy ( IMRT) has been proposed by scholars for cervical cancer patients presenting with large residual tumors accompanied by parametrial infiltration after IMRT. In this article, recent research progress on IMRT combined with brachytherapy applied in the treatment of cervical cancer was reviewed.