Objective To investigate the appropriate setting up of normal reference ranges of lymphocyte subsets in some flow cytometry laboratories and to study the effects of different flow cytometers and various reagents by different manufacturers on the analysis of peripheral blood lymphocyte subsets. Methods Three FCM labs (named A, B and C) in Beijing region were selected representing 3 commonly used flow cytometers (Beckman Coulter Epics XL, Beckman Coulter Cytomics FC500, BD FACS Calibur). 50 samples from healthy donors were distributed to 3 labs and tested according to individual lab's standard operating procedure to verify whether the normal reference ranges of peripheral blood lymphocyte subsets established were appropriate. The application of internal quality control was also investigated. Commercial blood quality control reagents were given to the 3 FCM labs and tested within 20 working days paralleled with routine samples. In addition, 20 patients' samples were prepared using 4 different combinations of reagents ( a , b , c and d). The results from combination a, which used the Beckman Coulter reagents and instrument, were compared to the results from combination b, c and d, which used reagents from different manufacturers. Then the prepared samples were tested on Beckman Coulter Epics XL to evaluate the effects of different combinations of reagents on the results of peripheral blood lymphocyte subsets analyzed by the same instrument. Furthermore, 24 patients' samples prepared by same reagents from Beckman Coulter company were tested on both Beckman Coulter Epics XL and BD FACS Calibur respectively to assess the effects of different instruments on peripheral blood lymphocyte subsets. 20 patients' samples prepared by same reagents and instruments were analyzed by Beckman Coulter Epics XL analytic system and BD FACS Calibur analytic system respectively to assess the effects of the two analytic systems on the lymphocyte subsets. Results Over 10% of the results for NK and T4/T8 in lab A as well as T4 in labs B and C fell outside of their normal reference ranges. The probabilities exceeding corresponding normal reference ranges were 16% ( 9/50 ), 24% ( 12/50 ), 22% (11/50) and 12% ( 6/50 ), respectively. The results using internal blood quality control in 3 FCM labs within 20 working days were all within the reference ranges of the quality control provided by the kit. The biases from b and c reagent combinations were substantial compared with that of reagent a combination. Among the biases from b and c reagent combinations, the lowest probability of bias exceeding 10% was T8 of combination b, which had probability of 70% (14/20). The highest probabilities of hias exceeding 10% were T3 and T4 of b and c reagent combinations, which reached 100% (20/20) . Furthermore, the biases of T3, T8 and B of d reagent combination compared with that of reagent a combination were also substantial. The probabilities of bias exceeding 10% were 35% (7/20) ,85% (17/20) and 75% (15/20), respectively. Comparing the results of samples prepared and analyzed by reagents and instruments from different manufacturers to that of samples prepared and analyzed by the same company's reagents and instruments showed that there were great discrepancies in T3, T4 , T8 , B and NK. The probabilities of bias exceeding 10% were 71% ( 17/24), 80% (19/24) ,38% (9/24), 33% (8/24) and 92% (22/24), respectively. The biases of T8, NK and B were substantial when compared the results from Beckman Coulter Epics XL analytic systems and BD FACS Calibur analytic systems. The probabilities of bias exceeding 10% were 55% (11/20 ), 70% ( 14/20 ) and 55% (11/20), respectively. Conclusions FCM labs should set up their own normal reference range for peripheral blood lymphocyte subsets. The normal reference range should be verified periodically. It is important to apply internal blood quality control regularly and accumulate the quality control results. The reagents and instrument for preparing peripheral blood samples should be from the same manufacturers.

Objective To compare postoperative adjuvant chemoradiotherapy with adjuvant chemotherapy in patients with gastric cancer by a meta-analysis.Methods PubMed,EMbase,Cochrane Library,Wanfang,CNKI,VIP,and CBM databases were searched to identify the controlled clinical trials of postoperative adjuvant chemoradiotherapy versus adjuvant chemotherapy for gastric cancer.The obtained data were analyzed using RevMan 5.2.5 and Stata 12.0.The difference between two groups was estimated by calculating the odds ratio (OR) with 95％ confidence interval (CI).Results A total of 12 controlled clinical trials involving 1674 gastric cancer patients,which were selected according to inclusion and exclusion criteria,were included in this meta-analysis.The meta-analysis showed that the 3-and 5-year survival rates were significantly higher in the adjuvant chemoradiotherapy group than in the adjuvant chemotherapy group (OR=2.96,95％ CI=1.75-5.03,P=0.000; OR=1.45,95％ CI=1.06-1.99,P =0.020);the local recurrence rate was significantly lower in the adjuvant chemoradiotherapy group than in the adjuvant chemotherapy group (OR =0.50,95％ CI =0.34-0.72,P =0.000) ; there was no significant difference in distant metastasis rate between the two groups (OR =0.79,95％ CI =0.58 -1.07,P =0.130).Conclusions The meta-analysis of existing study results shows that compared with adjuvant chemotherapy alone,adjuvant chemoradiotherapy is a relatively safe and effective postoperative treatment for gastric cancer.

ObjectiveTo explore the relative factors of the compliance in schizophrenic out-patients.Methods118 schizophrenic out-patients chosen randomly were investigate with the compliance and assessed with Clinical Global Impression-Severity of Illness (CGI-SI), Social Disability Screening Schedule (SDSS), Brief Psychiatric Rating Scale (BPRS) and Treatment Emergent Symptom Scale (TESS) by telephone. Logistic regression analysis was used to confirm the relative factors of the compliance.Results56.8% of all patients were of full compliance. The risk factors of compliance in schizophrenic out-patients were the frequency of medicine taken, the scores of CGI-SI and the insight of the illness. ConclusionIt is important to choose effective and convenient antipsychotic drugs and improve the insight of the illness to enhance the compliance of the schizophrenic out-patients.

Objective To retrospectively analyze of clinical application of BF-XP60 micro-bronchoscopy.Methods 135 clinical data of patients who adopted ultrafine micro-bronchoscopy and intervention were collected and analyzed for the complications.Results The frequency of local rhinomusoca damaging and errhysis was in 3 cases,the mucous of the glottis damaging and errhysis was in 2 cases,local mucous of the tracheal bronchus errhysis was in 3 cases.After intervention,the frequency of fever was in 13 cases,massive haemorrhage was in 1 case,pneumothorax was in 1 case,chest pain was in 2 cases,part fiber of inner untrafine micro-bronchoscopy broken was in 2 cases,check failure due to ultrafine micro-bronchoscopy broken in trachea was in 4 cases,and arrhythmia,asphyxia,and death were in 0 case.The overall incidence of side effects was 22.9％ (31/135).Conclusion Application of ultrafine micro-bronchoscopy was contributed to find the lesions within the bronchioles and around the lungs,moreover,it could evaluate the distal bronchus of airway obstruction which was planned to adopt intervention.The topic that how to reduce the incidence of the side effects of the micro-brohchoscopy and improve the success rate and safety of inspection and intervention was worth to be concerned.

Objective To retrospectively analyze the effect of two kinds of biological agents in volume -re-duced bullae .Methods 11 patients who suffered from bullae were operated under large C-arm locating ,and infused two kinds of biological agents through micro catheter of fibreoptic bronchoscopy .All of them were randomly divided into the two groups .The biological agents in group A were fibrinogen and diluent thrombin , and that of group B was Porcine Fibrin Sealant Kit .In group A,the micro catheter with diameter of micro thread less than 1.2mm was placed in bullae through fibreoptic bronchoscope ,and then the 2mL lidocaine,5 ml fibrinogen,and double of 500u diluent thrombin were inproperorder injected through micro catheter .In group B,the Porcine Fibrin Sealant Kit was injected at the same method,and then the suspension fluid was exacted .The operation time was recorded ,and then the clinical efficacy and incidence rate of complications were compared .Results The operation time of group A was 5-15 minutes, and that of group B was 6-20 minutes.For all the patients ,4 cases were totally effective ,2 cases were significantly effective,and 2 cases were totally non-effective.The total effective rate was 81.82%(9/11).The incidence rates of common complications in group A and B were 52.38%(22/42),58.33%(14/24),respectively,the difference was not significant (χ2 =0.22,P>0.05).Moreover,there were no serious complications in all cases .Conclusion The security and effect of two kinds of biological agents might be well enough ,but in view of less cases ,they were worth to further popularized and applied in clinical practice .

OBJECTIVETo observe cerebral protective effect of muscone (nasal administration) on traumatic brain injury model rats.

METHODSSD rats were divided into the sham-operation group, the model group, and the treatment groups according to random digit table, 50 in each group. Traumatic brain injury model was established by controlled cortical strike. Rats in the sham-operation group received surgery and anesthesia procedures only, with no strike. Muscone (1.8 mg/kg) was delivered to rats in the treatment group using in situ nasal perfusion, 30 min each time, twice daily for 7 successive days. Water content of brain tissue was detected in each group before intervention (T1), at day 3 of intervention (T2), day 5 of intervention (T3), and after intervention (T4), respectively. Expression levels of brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detected using immunohistochemical analysis.

RESULTSCompared with the sham-operated group, water content of brain tissue increased (P < 0.05), and expression levels of NGF and BDNF decreased in the model group at T1, T2, T3, and T4 (P <0. 01). Compared with the model group, water content of brain tissue decreased (P < 0.05), and expression levels of NGF and BDNF increased (P < 0.01) in the treatment group at T1, T2, and T3.

CONCLUSIONNasal administration of muscone could reduce water content of brain tissue, alleviate cerebral edema, promote secretion of BDNF and NGF by olfactory ensheathing cells in traumatic brain injury rats.

Animals ; Brain ; drug effects ; Brain Injuries ; drug therapy ; Brain-Derived Neurotrophic Factor ; metabolism ; Cycloparaffins ; pharmacology ; Nerve Growth Factor ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.

Objective To evaluate the biological variations of 8 lymphocyte subsets using flow cytometric double-platform method.Methods Twenty healthy adults were recruited from Peking Union Medical College Hospital in September 2013.At 8:00 AM,12:00 PM,and 4:00 PM on days 1,3,and 5,venous blood was collected from the volunteers.The percent and absolute lymphocyte subset counts were measured using duel-platform method.The sample collection and handling techniques were standardized.Before each batch analysis,the instrument quality controls were performed using the same lots of reagents.The intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were calculated by nested ANOVA with SPSS 13.0 software.The analytical coefficient of variation (CVA),index of individuality (Ⅱ) and reference change value (RCV) were calculated by Excel2003.A mean pairwise comparison was determined by one-way ANOVA and variance analysis.The values between groups were analyzed by independent sample t test.Results For T cells (CD3 +),helper T cells (CD3 + CD4 + CD8-),suppressor T cells (CD3 + CD4-CD8 +) and B cells (CD3-CD19 +),the intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were 0.03,0.06,0.05,0.14 and 0.12,0.16,0.23,0.31 respectively,which were all similar to those in previous studies.However,the Ⅱ and RCV of the four lymphocyte subsets were very different from those in previous studies,which were 0.26,0.40,0.22,0.44 and 8.77,16.86,14.93,39.69,respectively.Moreover,variations in absolute count,CVI,CVG,and analysis coefficient of variation (CVA) of all 8 lymphocyte subsets were greater than those of relative count.Variations in the percent and absolute counts for the CD3 + CD4-CD8-,CD3 + CD4 + CD8 +,and CD3+ CD16+ CD56+ cell subsets were relatively high.The CVI,CVG and CVA for the cells of CD3 + CD4-CD8-were 0.12,0.49 and 0.16.The CVI,CVG and CVA for the cells of CD3+ CD4+ CD8+ were 0.40,0.93 and 0.55.The CVI,CVG and CVA for the cells of CD3 + CD16 + CD56 + were 0.28,1.11 and 0.16.Conclusions Investigation on the CVI,CVG and CVA may allow us to obtain Ⅱ and RCV,by which we can determine the utility of traditional population based reference ranges.Documentation of the RCV indices may be used as objective delta-check values in quality management and decide whether clinical significance existed in the continuously detected results.

Objective To explore the security and its influencing factors on benign airway stenosis treated with interventional of high pressure balloon expansion catheter.Methods Clinial data of 39 cases of inpatients suffered from benign airway stenosis were chosen.17 cases were male,and 22 cases were female.The ages of them ranged from 15 to 83 years old.According to the clinical symptoms,HRCT 3D reconstruction,and the results of bron-choscope,all patients were treated with balloon expansion catheter at different criterions.The balloon catheter with size that slightly smaller than the targeted normal bronchial tube was chosen,expansion for average 1 -4 times,single balloon expansion time ranged from 0.5 to 4 min,the pressures were kept at 3 -6 atmosphere,and the highest pres-sure did not exceed 8 atmospheric pressure.The efficacy and complications were retrospectively analyzed.Results 19 cases were completely effective,14 cases were basically effective,6 cases were completely ineffective,and the total effective rate was 84.6% (33 /39 ),the incidence of complications was 35.8% (14 /39 ),moreover,no deaths occurred.Conclusion High pressure balloon catheter expansion is one of commonly used technology in breathing interventional treatment;it has the characteristics of easy operation,and immediate curative effect,and so on.But if the improper operation,incorrect selection of the case,or inaccurate evaluation of the stenosis during operation,serious complications and unnecessary iatrogenic injury can be occurred.Therefore,it is worthy of attention and further summarizing by breathing interventional physicians.

BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.