ObjectiveTo investigate the serum tumor markers level of carcinoembryonic antigen(CEA) ,cytokeratin fragment antigen21-1 (CYFRA21-1),neuron specific enolase (NSE) in patients with lung cancer, and the change after chemotherapy on them. Methods Radioimmunoassay was applied to detect the levels of CEA, CYFRA21-1 ,NSE in 45 patients with advanced NSCLC before and after chemotherapy,and the tumor markers were also detected in 20 patients with SCLC and 20 patients with benign lung diseases of control groups. ResultsThe levels of CEA, CYFRA21-1, NSE in lung cancer group before chemotherapy were much higher than benign group, but there was no difference of CYFRA21-1 between the SCLC group and benign group. The same result of NSE was found between NSCLC and benign group(P ＞0. 05). The value of NSE was lower in the patients with SCLC after chemotherapy than before(P ＜0. 01 ). The level of CYFRA21-1 was lower in squamous carcinoma than before( P ＜0. 01 ). But in the adenocarcinoma group only NSE's level was lower after chemotherapy( P ＞0. 05) ,there were no differences in CEA and CYFRA21-1 ( P ＞ 0. 05 ). ConclusionThe levels of the three tumor markers rise obviously in advanced NSCLC and decrease after chemotherapy. The differences were significant with NSE in SCLC and CYFRA21-1 in squamous cell carcinoma and CEA in adenocarcinoma. The levels of serum CEA,CYFRA21-1 and NSE could be a tumor marker in progressive lung cancer. And the decrease of the levels could be used to evaluate the chemotherapeutic response respectively in different pathologic types of lung cancer.

Objective To study the microcirculation and structural changes, surviving area of expanded prefabricated flaps. Methods A total of 40 New Zealand rabbits were divided randomly into expanded prefabricated, expender lined, simple prefabricated and free flap groups, each consisting of 10 rabbits. For the expanded prefabricated, expender lined and simple prefabricated groups, after the femoral artery and vein were transplanted into subcutaneous tissues of abdomen, and expanders were implanted into the deeper dartos. The free flap group was a blank control group. For the expanded prefabricated group, the expansion was carried out on 7th day postoperatively. On postoperative day 52, when the expander was fully expanded, island flaps with the prefabricated vessels as the pedicles were formed. The flaps were measured by laser Doppler flowmetry, light microscopy and digital re-cording of survival arca. Results When compared with the other groups, the perfusion volume of mi-crocirculation enhanced, flaps survival improved (97.54±2.73) %, blood capillary were stronger, to-gether with microscopic changes were significant in the expanded prefabricated groups (P<0.05). Conclusion Expandedprefabricated flaps can increase the survival size of the flaps and the safety of flap transplantation.

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.

Objective To evaluate the immune reconstitution of HIV-1 infected individuals after long-term highly antiretroviral therapy (HAART). Methods Twenty-five HIV-1 infected individuals receiving HAART, 17 without HAART and 15 healthy controls were included in the study. CD4 +T, CD8 +T,CD8/human leukocyte antigen DR (HLADR) + T, CD8/CD38 +T cells, and the expression of CD127 on CD3 +T cells from peripheral blood samples were measured by flow cytometry. IL-7 in peripheral blood was measured by enzyme-linked immunosorbort assay (ELISA) in HAART group. t test was performed to compare the measurement data among the groups. Results Before HAART, the count of CD4 + T cells in HIV-1 infected group was lower than that of the healthy control (t =9. 12, P ＜0. 01 ), while the counts of CD8 + T,CD8/HLADR+T, and CD8/CD38 +T cells were higher than those of the healthy control (t = 4.48, 4.89 and 3.88, P＜0. 01 ). Ater 7 years' antiviral therapy, CD4 +T cells increased, CD8 +T cells decreased, but both of them didn' t reach to the normal levels ( t = 2.66 and 2.43, P ＜ 0.05 ). While the counts of CD8/HLADR+T cells and CD8/CD38 +T cells almost reached to the normal levels (t = 0. 86 and 1.39, P ＞0.05). Before HAART, the concentration of IL-7 in HIV-1 infected group was higher than that in healthy controls (t =5.31, P ＜0.01 ). It decreased with HAART, but was still higher than the normal level (t =2. 81, P ＜ 0. 05 ). The expression of CD127 on CD3 + CD8 + T cells in non-HAART group was significantly lower than that in healthy control ( t =- 6.01, P ＜ 0.01 ), while that in HAART group was higher ( t = 2.32,P ＜0.05), but still not reached to the normal level ( t = 4.49, P ＜ 0. 05 ). CD127 expression on ( CD45RA + ) CD3 + CD8 + T cells almost increased to the normal level ( t= 0. 28, P ＞ 0. 05 ), while that on ( CD45RO + ) CD3 + CD8 + T cells was still remarkably lower than the normal ( t = 4. 86, P ＜ 0. 05 ). Conclusion Long-term HAART can partially restore the count and function of lymphocyte subsets in HIV-1 infected individuals, and the abnormal immune activation can be inhibited.

Objective To study the changes and significance of CD4+CD25+ regulatory T cells and C-reactive protein(CRP) in patients with acute exacerbation of chronic obstructive pulmonary disease(COPD). Methods Flow cytometry was used to detect the frequency of CD4+CD25+ regulatory T cells in peripheral blood from 36 patients with acute exacerbation of COPD( COPD group) and 36 patients with clinical stability of COPD(control group one)and 36 normal individuals(control group two). The level of CRP was detected routinely. Results The ratio of CD4+CD25+ regulatory T cells number in peripheral blood of COPD group to the total number of CD4+T cell was (2.56±1.83 )%, and it was significantly decreased compared to the other two groups (P all<0.01 ). The level of CRP in COPD group was markedly higher than that in the other two groups (P all<0.01 ). The level of CD4+CD25+ regulatory T cells in patients with acute exacerbation of COPD had negative relation with CRP. Conclusions CD4+CD25+ regulatory T cells participate in inflammatory response. The proportion of CD4+CD25+ regulatory T cells decreases in patients with acute exacerbation of COPD, and it may result in maladjustment of cytoimmunity.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

OBJECTIVE To optimize the extraction technology for the raw drugs of Sanse powder gel paste. METHODS SD rats were divided into blank group， model group， traditional technology group， water extraction group and ethanol extraction group， with 5 rats in each group. Anterior cruciate ligament transection was used to construct knee osteoarthritis model， and the pharmacodynamic effects of different extraction methods on arthritic rats were investigated. Analgesic experiments were conducted using cold and hot pain thresholds and pain mediators calcitonin gene-related peptide （CGRP）， cyclooxygenase-2 （COX-2）， substance P （SP）， and prostaglandin E2 （PGE2） contents as indicators. HE staining was performed on the synovial membrane of rats to observe the degree of synovial cell proliferation， inflammatory infiltration and vascular invasion， and anti-inflammatory experiments were conducted using protein and mRNA expressions of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and IL-6 as indicators. The analgesic and anti-inflammatory effects were compared among those groups. In the orthogonal test， ethanol dosage， extraction time and extraction times were used as evaluation factors， and the contents of casticin， strychnine and toxiferine were taken as evaluation indicators； comprehensive score was calculated. The validation experiments were carried out after optimizing the extraction technology of the raw drugs of Sanse powder gel paste. RESULTS Compared with the model group， the cold and heat pain thresholds of drug administration groups （except for the traditional technology group） were all increased significantly （P＜0.05）， while the contents of pain （No.Y2021rc02） mediators CGRP， COX-2， SP and PGE2 were all decreased significantly （P＜0.05）. HE staining showed that inflammatory cell infiltration， fibrosis and collagen deposition were 炎。E-mail：liuzixiu3221@126.com decreased in the administration groups； a small amount of capillary proliferation could be found； the protein and mRNA expressions of inflammatory factors such as IL-1β， IL-6 and TNF-α were decreased significantly in synovial tissue of rats in administration groups （P＜0.05）. Compared with the traditional technology group， most indicators of the ethanol extraction group were significantly reduced （P＜0.05）， and only heat pain threshold and mRNA expression of IL-6 in rats were decreased significantly in the water extraction group （P＜0.05）. The optimal extraction technology of the raw drugs of Sanse powder gel paste included suitable dose of Sanse powder， 8-fold 55% ethanol， heating reflux extraction for 90 minutes， extracting twice. The results of 3 times of verification experiments showed that the average contents of casticin， strychnine and toxiferine were 0.007%， 0.092%， and 0.214%， respectively； RSD were all less than 5%. CONCLUSIONS The optimized extraction technology for the raw drugs of Sanse powder gel paste is stable and feasible， which can improve the efficacy of the preparation.

Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Retrospective Studies