Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.

Objective To investigate the effects of high spermatic vessel dissection on testicular morphological alteration of SD rats in prepuberty,puberty and sexual maturity phases.Methods Thirty-day-old SD rats were divided into 2 groups underwent sham operation and left high spermatic vessel dissection as a simulation of Palomo′s maneuver.Detailed morphological investigations were made at 3 different postoperative intervals among the 3rd day,30th day and 56th day.Results High spermatic vessel dissection in prepubertal rats induced acute testicular ischemia in the operated testes on the 3rd day.Most of the operated testes on the 30th day showed testicular atrophy.And all the operated testes showed testicular atrophy and sperm disappearance in epididymis on the 56th day.Conclusion High dissection of spermatic vessel in prepubertal rats induced testicular ischemia in prepuberty and testicular growth failure in puberty,testicular atrophy completely and sperm production losing in sexual maturity phase.

OBJECTIVETo investigate the impact of PET/CT on the clinical staging, target volume delineation and precise radiotherapy (PAR) planning for patients with non-small cell lung cancer (NSCLC).

METHODSPET/CT scanning was performed in 58 histologically proven NSCLC patients for radical radiotherapy or surgery. The clinical staging of all patients was determined by PET/CT according to 1997 World Health Organization (WHO) staging system. The gross tumor volume (GTV) was delineated and three-dimensional conformal radiotherapy (3D-CRT) planning was established with identical parameters based on CT image and PET/CT fused image, respectively. The indexes including volume of GTV (V(GTV)), percentage of the total lung volume which received more than 20 Gy (V(20)), mean lung dose (MLD), tumor control probability (TCP), normal tissue complication probability (NTCP), and dose to spinal cord (Ds) were selected and evaluated. The quality of the two plans and the impact of PET/CT on PAR planning was compare and analyzed.

RESULTS1. PET/CT image results changed the clinical stages in 21 of 58 (36.2%) patients with 14 upstaged and 7 downstaged, therefore, the management decisions were modified in 16 (27.6%) patients. 2. Among 32 patients who underwent surgery, PET/CT staging result was consistent with pathologic staging in 29 with one false negative and 2 false positive in lymph node staging. The sensitivity of PET/CT was 96.9% and accuracy 90.6%. 3. The differences of indexes including V(GTV) (P = 0.004), V(20) (P = 0.000) and MLD (P = 0.004) between the two radiotherapy plannings were statistically significant, whereas, the Ds, TCP and NTCP (left lung, right lung, skin and spinal cord) was not.

CONCLUSION1. The impact PET/CT on clinical staging of NSCLC and PAR planning was remarkable. 2. PET/CT is more consistent with pathology in staging than CT and, therefore, is an important compensatory staging measure. 3. Compared with CT, PET/CT can reduce the V(GTV) in patients with atelectasis and obstructive pneumonitis when contouring the target volume, so can provide better protection for normal surrounding lung tissue. On the other hand, PET/CT is more sensitive in detecting mediastinal lymph node metastasis than CT, and the V(GTV) can be more precise and guaranteed. 4. Radiopneumonitis may be more effectively prevented because of significant decrease in V(20) and MLD by more precise planning based on PET/CT results. 5. PET/CT not only can provide satisfactory Ds, TCP and NTCP within clinical demand, but also more precise delineation of the radiation target volume and precise radiotherapy planning for NSCLC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Female ; Humans ; Lung Neoplasms ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Positron-Emission Tomography ; methods ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Conformal ; methods ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the safety and efficacy between endoscopic papillary balloon dilatation (EPBD) and endoscopic sphincteropapillotomy ( EST) for common bile duct stones using meta-analysis method.

METHODSRandomizd controlled trials comparing EPBD with EST for common bile duct stones and published from January 1990 to July 2012 were recruited. This meta-analysis was conducted to estimate short-term and long-term complications. Fixed random effect model or random effect model was established to analyze the data.

RESULTSTwelve randomizd controlled trials were included in this analysis. These studies included 1865 patients, 925 of them were treated with EPBD and 940 were treated with EST. The analysis of basic characteristics of these included studies showed that: compared to EST, patients in the EPBD group were younger (OR = -1.16, 95% CI: -1.49 to -0.84, P = 0.00), while in two groups, there were no significant difference (P > 0.05) in gender proportion, average size of stones, number of gallstones, previous cholecystectomy, the number of merged duodenal diverticulum, common bile duct diameter, the total follow-up time. Also, compared to EST, the overall stone clearance in the EPBD group was lower (OR = 0.64, 95% CI: 0.42 to 0.96, P = 0.03), pancreatitis incidence was higher (OR = 2.67, 95% CI: 1.61 to 4.43, P = 0.00), incidence of bleeding (OR = 0.12, 95% CI: 0.04 to 0.34, P = 0.00), acute cholecystitis (OR= 0.39, 95% CI: 0.18 to 0.84, P = 0.02), total long-term complication rate (OR = 0.53, 95% CI: 0.36 to 0.77, P = 0.01), stone recurrence rate more than a year were lower (OR= 0.48, 95% CI: 0.26 to 0.90, P = 0.02). While in two groups, there were no significant difference (P > 0.05) in the stone removal on 1 '' attempt, the total near-term complications and acute cholangitis.

CONCLUSIONSOn the basis of lower rates of bleeding, EPBD seems to be preferred strategy over EST for endoscopic remove of common bile duct stones in patients who have coagulopathy. Although stone recurrence rate more than a year of EPBD is lower, but the overall stone clearance rate is lower and the risk of pancreatitis is higher than that of EST.

Dilatation ; Gallstones ; surgery ; Humans ; Postoperative Complications ; epidemiology ; Randomized Controlled Trials as Topic ; Sphincterotomy, Endoscopic ; Treatment Outcome

Objective To study the effects of sleep inertia (SI) of different time course sleeps on sleep stages and cognitions in healthy men after 30 h sleep deprivation,and also to investigate the mechanism of cognitive functions impairment in sleep inertia stages and the influential factors of sleep inertia.Methods Ten healthy men (age,(20.8 ±2.1) years) participated in the program.The program was divided into 2 stages.First,participants attended a series of tests after 20 min nap(20 min nap group)after 30 h sleep deprivation.The testing series included 3 parts:the continuous performance task,the Stroop Tests,and the Addition Tests.The series of tests were done 3 times immediately after the volunteers were awoken and each lasted about 15 minutes with an interval of 10 minutes between each test.The polysomnogram (PSG) was recorded during the nap.The following 7 days was set as washing-out period to ensure a complete recovery.Participants repeated the similar processes with 2 h nap(2 h nap group) instead of 20 min nap.The cognitive performance of each group was compared with each other along with the best cognitive performance in awakening to estimate whether or not the cognitive abilities regained the normal condition.Results ( 1 ) Sleep latency became shortened in both groups after 30 h sleep deprivation.There were no slow wave sleep (SWS) and rapid eye movement sleep (REM) sleep stages in the 20 min naps,while the percentage of SWS was increased and percentage of REM declined in 2 h naps.(2)In the early of SI (5 min after awaking),cognitive tasks showed that the abilities of continuous attention,selected attention and addition ability were all impaired (continuous performance task:(371.8 ± 21.3 ) times/3 min vs (334.4 ± 22.4) times/3 min,( 373.2 ± 19.0) times/3 min vs ( 323.7 ± 23.8) times/3 min,t =10.443,7.774,both P＜0.01; Stroop tests:(20.3 ±1.5) points vs(17.3 ± 1.0) points,(21.5 ±0.8)points vs( 16.1 ± 1.4 ) points,t =8.478,4.934,both P ＜ 0.05 ; Addition Tests:( 222.2 ± 13.2 ) s vs ( 266.6 ±23.7 ) s,( 226.3 ± 10.9) s vs ( 267.6 ± 23.4 ) s,t =5.748,6.685,both P ＜ 0.01 ).The cognitive functions impairments of 2 h nap group were more severe at the initiation of sleep inertia,but regained the normal condition more quickly.Different cognitive tasks recovered at different speeds.The dispersion of SI needed 30 min.Conclusions ( 1 ) There are difference in the sleep construction and awaked sleep stage between 20 min nap and 2 h nap groups.(2) SI exerts negative influences on cognitive performances of continuous attention,selected attention and addition after sleep deprivation.Many factors may influence the dispersion of SI,including sleep debt,compensation of sleep debt and others.(3) Fragments of sleep are unfavorable to the recovery of body.

The extraction of active ingredient group, i.e., prim-o-glucosylcimifugin, astragaloside, and 5-o-methylvisammiosode from Yupingfeng powder with one-pot method was studied in this work. A HPLC method was used to determine the content of each active constituent mentioned above in the extract. The influences of extraction temperature, time, volume percent of ethanol in water and its amount added on the content and yield of active ingredient group were investigated by orthogonal test. The experimental results showed that the optimized extraction conditions were as follows: 1g of Yupingfeng powder was one-pot extracted for 4 hours at 80 degrees C with 90% ethanol as solvent, and the yield and content of active ingredient group were 0.16%, 0.53% respectively. The active ingredient group in Yupingfeng powder could be effectively one-pot extracted under the conditions above.
Apiaceae ; chemistry ; Astragalus membranaceus ; chemistry ; Atractylodes ; chemistry ; Chemical Fractionation ; methods ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Monosaccharides ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; Powders ; Saponins ; analysis ; isolation & purification ; Temperature ; Triterpenes ; analysis ; isolation & purification ; Water ; chemistry ; Xanthenes ; analysis ; isolation & purification

OBJECTIVETo observe the difference in the efficacy on the symptoms of bronchial asthma at the chronic persistent stage between acupoint heat-sensitive moxibustion and western medicine with Seretide.

METHODSSixty-four cases were randomly divided into a heat-sensitive moxibustion group (32 cases) and a western medication group (32 cases). In the heat-sensitive moxibustion group, the sensitized points located between Feishu (BL 13) and Geshu (BL 17) or in the region 6-cun lateral from the 1st and the 2nd intercostal spaces of the chest were selected. The heat-sensitive moxibustion was adopted, continuously for 8 days, once per day. In the later 22 days of the 1st month, 12 treatments should be ensured. Two months later, 15 treatments should be guaranteed each month. The time of each treatment was 30 to 90 min. Totally 50 treatments were required. In the western medication group, Seretide inhaler was adopted, one inhalation each time, twice per day, for 3 months totally. The asthmatic symptoms were scored for the patients in two groups and the comparison was made between the two groups.

RESULTSAfter 3 months of treatment, the asthmatic symptom scores were all improved for the patients in the heat-sensitive moxibustion group and the western medication group as compared with those before treatment (both P < 0.05). In 6 months of follow-up visit, the asthmatic symptom scores in the heat-sensitive moxibustion group were stable, but those in the western medication group were reduced, there was significant difference between the two groups (P < 0.05).

CONCLUSIONThe acupoint heat-sensitive moxibustion effectively relieves the clinical symptoms for the patients with bronchial asthma at the chronic persistent stage. Its efficacy is similar to that of Seretide inhaler. But the long-term efficacy of the heat-sensitive moxibustion is much better.

Acupuncture Points ; Adolescent ; Adult ; Albuterol ; analogs & derivatives ; therapeutic use ; Androstadienes ; therapeutic use ; Asthma ; therapy ; Chronic Disease ; Drug Combinations ; Female ; Fluticasone-Salmeterol Drug Combination ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Moxibustion ; methods

Objective　To investigate the effect of rapid fluid replacement on hemorheology in delayed resuscitation after burn. Methods　A total of 32 dogs inflicted with 40%TBSA full thickness scalding were randomly divided into 4 groups: scald control group(C group), delayed Gelofusion even replacement group (GE group), rapid fluid replacement group (GR group), and delayed plasma rapid fluid replacement group (PR group). The femoral arterial pressure, viscosity of blood and plasma, packed cell volume and aggregation of RBC were detected at the intervals of before and 2, 6, 8, 12, 24, 36 and 48 hours after scalding. Results　The viscosity of blood markedly increased at 2 hours after scalding, and the hemorheology parameters decreased after fluid resuscitation. The hemorheologic parameters were obviously lower in GR group than in GE group at 2 hours after rapid resuscitation, the viscosity of blood and RBC aggregation in GR group were obviously lower than those in PR group. Conclusion　Under the condition of delayed resuscitation after burn, rapid fluid replacement can quickly decreased the state of blood high viscosity and may play a role in improving microcirculation and treating burn shock.

OBJECTIVETo explore the prognosis factors of hilar cholangiocarcinoma, and investigate the relation between operative procedure and prognosis of it.

METHODSA retrospective cohort study was investigated in 198 patients with hilar cholangiocarcinoma, who were treated in our hospital from December 1997 to December 2002. There were 117 males and 81 females. The age ranged from 27 to 81 years old with a mean of 56. Jaundice (94.5%), pruritus (56.6%) and abdominal pain (33.8%) were the main present symptoms. According to Bismuth-Corlette classification, there were 14 type I cases, 19 type II cases, 12 type IIIa cases, 15 type IIIb cases, 112 type IV cases and 26 unclassifiable cases. One hundred and forty four cases received open operative treatment, and the others only were treated with endoscopic approach (including ERBD or EMBE 21 cases, ENBD 31 cases) or percutaneous transhepatic cholangiodrainage (2 cases). Tumor resection was performed on 120 cases with a resection rate of 83.3%, included radical resection 59 cases (41.0%). Twenty-four cases underwent paunched biliary exploration and drainage.

RESULTSThe Cox's regression model analysis showed that occupation, preoperative maximum total serum bilirubin level, operative procedure and postoperative adjuvant radiation affected postoperative survival significantly, but gender, age, choledocholithiasis, hepatitis, preoperative serum CA19-9 level, Bismuth-Corlette type, histopathologic grading and postoperative chemotherapy were not significant prognostic factors. The postoperative survival of biliary drainage group, palliative resection group and radical resection group, which statistically differed pairwise. Between ERBD or EMBE group and palliative resection group, there was no statistical difference. So was between ERBD or EMBE group and biliary drainage group, or between ENBD group and biliary drainage group. The survival differed statistically between ERBD or EMBE group and ENBD group.

CONCLUSIONSOperative procedure was the most important prognosic factor of hilar cholangiocarcinoma, radical resection still was the primary measure to cure and long term survival. For irresectable hilar cholangiocarcinoma, the effect of ERBD or EMBE could not be considered to be worse than that of open operative treatment.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; surgery ; Bile Ducts, Intrahepatic ; surgery ; Biliary Tract Surgical Procedures ; methods ; Cholangiocarcinoma ; surgery ; Drainage ; methods ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

BACKGROUNDHomoharringtonine (HHT) is effective in treating late stage chronic myelogenous leukaemia (CML), but little is known about long term maintenance during complete cytogenetic response. Long term efficacy and toxicity profiles of low dose HHT were evaluated in this study.

METHODSOne hundred and six patients with CML received 1.5 mg/m(2) of HHT alone by continuous daily infusion for seven to nine days every four weeks. Of 79 patients in the control group, 31 were treated with interferon alpha (IFN-alpha) and 48 with hydroxycarbamide. For 17 patients who failed to achieve cytogenetic response within 12 months' treatment of IFN-alpha, HHT was administered. Quantitative RT-PCR was used to detect the BCR-ABL mRNA expression in 36 Philadelphia positive CML patients enrolled after 2007. Haematological and cytogenetic responses were evaluated in all patients at the 12th month of follow-up. Long term efficacy was assessed in a follow-up with a median time of 54 months (12 months-98 months).

RESULTSAfter 12 months of therapy, cytogenetic response rate of the HHT, IFN-alpha and hydroxycarbamide groups were 39/106, 14/31 and 3/48, and corresponding molecular cytogenetic response rates 6/18, 3/8 and 0. Of the 17 patients who received HHT as salvage treatment, 6 achieved cytogenetic response (3 major). At the 48 months' follow-up, cytogenetic response was maintained in 32/39 patients treated with HHT. Patients who had cytogenetic response in HHT group or treated with IFN-alpha also showed longer median chronic durations, which were 45 months (12 months-98 months) and 49 months (12 months-92 months) respectively, indicating a longer survival time.

CONCLUSIONSLow dose HHT alone showed considerable short term and long term efficacy in the treatment of late stage CML. It may also be a good choice for patients who have failed imatinib, IFN-alpha treatment or haematopoietic stem cell transplantation or cannot afford these treatments.

Adolescent ; Adult ; Aged ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Female ; Fusion Proteins, bcr-abl ; genetics ; Harringtonines ; therapeutic use ; Humans ; Interferon-alpha ; therapeutic use ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Treatment Outcome ; Young Adult