BACKGROUND: Talomerase activity inhibitor inhibits or kills renal carcinoma cells, and also affects stem cells that play importan roles in occurrence and development of renal carcinoma.OBJECTIVE: To observe renal carcinoma stem cell surface marker CD133 and telomerase activity expression in serum-free suspension culture, and to compare with renal carcinoma cells in serum suspension culture.DESIGN, TIME AND SETTING: The in vitro cytological study was performed at the Jiangsu University from June 2008 to Februar 2009.WIATERIALS: Fresh normal renal tissue surrounding renal carcinoma was obtained from Affiliated Hospital, Jiangsu University.Renal carcinoma stem cell line OS-RC-2 was supplied by Cell Bank, Chinese Academy of Sciences Shanghai Branch.METHODS: OS-RC-2 in logarithmic phase, digested by trypsin, and centrifuged. Supematant was removed. OS-RC-2 cell line in serum-free DMEM/F12 supplemented with epidermal growth factor and basic fibroblast growth factor was incubated at 2×105/L in 5% CO2 incubator at 37℃. Renal carcinoma cultured in serum and normal renal tissue served as controls.MAIN OUTCOME MEASURES: Cell growth was observed under an inverted microscope. Expression of CD133 and CD34 was detected using flow cytometry. Reel-time quantitative TRAP assay was applied to evaluate telomerase activity in renal carcinoma stem cells.RESULTS: After incubated in serum-free medium, renal carcinoma stem cells were round and suspended. Two days later, cell mass generated. Each cell mass contained 3-8 cells, with strong refraction. Seven days later, cell mass became more, presented big body that was regular, round or elliptical. CD133+CD34- rate in renal carcinoma stern cell mass was significantly greater in serum-free suspension culture compared with in serum suspension culture. CD133 and CD34 expression was not determined in normal renal tissue. There were significant differences among groups (F=328.25, P < 0.05). Telomerase activity was greater in renal carcinoma stem cells and renal carcinoma cells compared with normal renal ceils (F=-278.74, P < 0.05). No significant difference was detected between renal carcinoma stem cells and renal carcinoma cells.CONCLUSION: Compared with serum cultured renal carcinoma cells, serum-free cultured renal carcinoma cell surface marker CD133 presents high expression. Moreover, talomerase activity is high in renal carcinoma stem cells and renal carcinoma cells compared with normal renal tissue.

Objective To evaluate the MUC1 promoter's role in driving gene expression in pancreatic cancer and its therapeutic significance.Methods Two plasmids were made.The plasmid pEGFP-MUC1N1 contained MUC1 promoter fragment connected to the pEGFP-N1 vector with the EGFG reporter gene.The pShuttle-MUC1-EGFP plasmid contained MUC1 promoter fragment and EGFP reporter gene connected to pShuttle plasmid.Lipofectamine 2000 was used to transfect the two plasmids into cells of MUC1-positive human pancreatic cell line Panc-1 and MUC1-negative human cervical carcinoma Hela.Fluorescence microscopy and flow cytometry compared the specificity and activity of the MUC1 promoter and CMV promoter.Results Reporter gene EGFP-positive cells 48 hours after transfection with pEGFP-MUC1-N1 and pShuttleMUC1-EGFP plasmid were 69.6％ and 63.6％ respectively,in Panc-1 cells,and 4.2％ and 3.7％ respectively,in Hela cells.Conclusions MUC1 promoter can drive reporter gene activity in MUC1-positive tumor cells targeting functional expression.There is potentially a use of targeted therapy in pancreatic cancer at the genetic level.

Objective: To investigate the delay in identification of pulmonary tuberculosis and influencing factors among children and adolescents in Jiaxing City, Zhejiang Province from 2013 to 2022, so as to provide the reference for targeted prevention and control measures. Methods: The information of pulmonary tuberculosis patients in Jiaxing City from 2013 to 2022 were captured from the Tuberculosis Information Management System of Chinese Disease Control and Prevention Information System, including demographics, diagnosis, treatment and etiological results. The delay in identification of pulmonary tuberculosis was analyzed among children and adolescents, and the factors affecting the delay in identification of pulmonary tuberculosis were identified using a multivariable logistic regression model. Results: A total of 2 407 pulmonary tuberculosis cases were reported among children and adolescents in Jiaxing City from 2013 to 2022, including 1 522 males (63.23%). The median age was 21.00 (interquartile range, 4.00) years. There were 410 students (17.03%), and 1 856 cases with non-local household registration (77.11%). There were 596 cases with delay in identification of tuberculosis (24.76%), 895 cases with delay in healthcare-seeking (37.18%) and 128 cases with delay in definitive diagnosis (5.32%). Multivariable logistic regression analysis that children and adolescents who occurred symptoms in the first quarter (OR=1.684, 95%CI: 1.261-2.249), were diagnosed first in county-level medical institutions (OR=3.800, 95%CI: 2.898-4.983) and had positive results of etiological testing (OR=1.534, 95%CI: 1.255-1.874) were more likely to delay in identification of pulmonary tuberculosis. Conclusions The delay in identification of pulmonary tuberculosis is associated with the time of symptom onset, the level of medical institution making first diagnosis, and the results of etiological testing. It is suggested to reinforce the publicity of pulmonary tuberculosis prevention and control, expand the coverage of screening programs and improve the diagnosis capability of medical institutions.

Objective: To investigate the hepatitis C prevention and control knowledge among clinicians in Jiaxing City, Zhejiang Province, so as to provide the evidence for intensified training and improved diagnosis and treatment of hepatitis C among clinicians. Methods: In November, 2021, clinicians were sampled using a stratified random sampling method from a city-level and a county (district)-level hepatitis C designated hospital in Jiaxing City. A questionnaire survey was performed using the Questionnaire for Hepatitis C Prevention and Control Knowledge among Clinicians, and the awareness of basic knowledge, professional knowledge and related knowledge about hepatitis C prevention and control among clinicians were descriptively analyzed. Results: A total of 186 questionnaires were allocated and 179 valid questionnaires were recovered, with an effective recovery rate was 96.24%. The respondents included 107 men (59.78%) and 72 women (40.22%) and had a mean age of (37.06±9.46) years. There were 107 respondents with a bachelor degree (59.78%), 56 with junior professional titles (31.28%), and 170 from non-infectious disease departments (94.97%). The awareness of basic hepatitis C prevention and control knowledge was 96.09%, and the awareness of “Transfusion of blood containing hepatitis C virus may acquire hepatitis C” was high (98.88%), and the awareness of “Hepatitis C can be cured” was low (77.09%). The awareness of professional hepatitis C prevention and control knowledge was 3.91% to 100.00%, and the awareness of “Pathogens of hepatitis C” (100.00%) and “Recommended screening populations for hepatitis C” (86.59%) was high, while the awareness of “There are two categories of hepatitis C cases: clinically diagnosed cases and confirmed cases” (3.91%) and “Clinical diagnosis of hepatitis C: positive anti-HCV antibody + any one of abnormal liver function or epidemiological history or clinical symptoms” (3.91%) was low. The awareness rates of “The state has included antiviral agents against hepatitis C into medical insurance” was and “Antiviral agents against hepatitis C are reimbursed in outpatient and inpatient departments of our hospital” were 81.56% and 59.78%, respectively. There were 69 clinicians participating hepatitis C-related training within one year (38.55%), and the awareness of clinicians that had participated in hepatitis C-related training had a higher awareness rate of basic hepatitis C prevention and control knowledge than those without participation (100.00% vs. 93.64%, P<0.05). Conclusion The awareness of basic hepatitis C prevention and control knowledge is high among clinicians in Jiaxing City; however, the training on diagnosis and classification criteria of hepatitis C and related medical insurance policy require to be improved.

Twenty-four soil samples of eight ecosystem-types around the Yellow River source area were investigated for the number and specific composition of soil dematiaceous hyphomycetes by dilution plate technique. And then the co-relationship between genus species of soil dematiaceous hyphomycetes and ecosystem-types was analyzed. The results show that the amount and species distribution of soil dematiaceous hyphomycetes had an obvious variability in different ecosystem-types, and that the dominant genus species varied in the eight ecosystem-types studied, with Cladosporium being the dominant genus in seven of the eight ecosystem-types except wetland. The index of species diversity varied in different ecosystem-types. The niche breadth analysis showed that Cladosporium had the highest niche breadth and distributed in all ecosystem-types, while the genera with a narrow niche breadth distributed only in a few ecosystem-types. The results of niche overlap index analysis indicated that Stachybotrys and Torula, Doratomyces and Scolecobasidium, Cladosporium and Chrysosporium had a higher niche overlap, whereas Arthrinium and Gliomastix, Phialophora and Doratomyces, Oidiodendron and Ulocladium had no niche overlap.
China ; Ecosystem ; Fresh Water ; microbiology ; Mitosporic Fungi ; classification ; isolation & purification ; Soil Microbiology ; Species Specificity

Objective To investigate clinical and epidemiological features of pneumocystis jirovecii pneumonia (PJP) in kidney transplant recipients. Methods Clinical data of 68 kidney transplant recipients admitted from July, 2021 to December, 2021 were collected. All patients were divided into the PJP group (n=11), common pulmonary infection group (n=24) and non-pneumonia group (n=33) according to the status of pulmonary infection. The incidence and treatment of PJP after kidney transplantation were analyzed. Basic characteristics and laboratory parameters of the recipients were compared among all groups. The genotyping and transmission map of PJP patients were evaluated. Results Among 64 kidney transplant recipients, 11 cases were definitely diagnosed with PJP. The most common clinical manifestations included elevated body temperature, and dry cough complicated with progressive dyspnea. Chest CT scan showed diffuse interstitial inflammation and ground glass-like lesions of bilateral lungs in all patients. After diagnosis, all patients were orally given with compound sulfamethoxazole for 3-4 weeks. Two patients received non-invasive ventilator-assisted ventilation due to severe lung infection and dyspnea, and the remaining patients were given with nasal cannula oxygenation. One patient experienced elevated serum creatinine level upon discharge, and developed renal allograft failure. The remaining 10 recipients with PJP obtained normal renal allograft function, and no recipient died. Compared with the non-pneumonia group, the rejection rate was higher, the length of hospital stay was longer, the lymphocyte count was less, the lymphocyte proportion was lower, the levels of C-reactive protein, serum creatinine and lactate dehydrogenase were higher, and the levels of serum albumin was lower and CD4⁺T cell count was less in the PJP group (all P < 0.05). Compared with common pulmonary infection group, the lymphocyte count was less, the lymphocyte proportion was lower, the CD4⁺T cell count was less and 1, 3-β-D- glucan (BDG) level was higher in the PJP group (all P < 0.05). No new genotype was detected in 10 of the 12 testing samples. It was considered that PJP mainly depended on two transmission chains and two independent transmission individuals. Conclusions Kidney transplant recipients are prone to pneumocystis jirovecii (PJ) infection due to impaired cellular immune function. The most common clinical manifestations consist of elevated body temperature and dry cough complicated with progressive dyspnea, accompanied by headache and fatigue in partial patients. Chest CT scan shows diffuse interstitial inflammation and ground glass-like lesion of bilateral lungs. PJ may be transmitted through respiratory tract. Small-scale PJP might occur in the follow-up outpatient of kidney transplant recipients. Preventive measures should be delivered in a timely manner.

Objective To develop a simple and effective method for monitoring cortical ischemia after temporary occlusion of the parent arteries during anterior circulation intracranial aneurysm surgery. Methods Fifty-two patients with anterior circulation aneurysm (58 aneurysms) received craniotomy from April to November 2008, and at the same time,cortical electroencephalograpby (EEG) and scalp EEG were monitored during the surgery.According to the international 10/20 electrode placement system, scalp electrodes were placed on O1, O2, P3, P4, T5, and T6 for monitoring the changes in the depth of anesthesia. A cortical strip electrode was placed on the cortical surface supplied by the artery that was possibly blocked during the operation, which was used to monitor the possible cortical ischemia. For patients who had cortical EEG suppression after the temporary occlusion of the parent arteries Were compared with the changes of scalp EEG. Whether there were ischemic events in the corresponding supply territory after vascular occlusion were observed after surgery. Results Of the 58 aneurysms, 40 aneurysms and 41 major arteries were occluded temporarily. After being occluded temporarily in 19 arteries of 18 patients, cortical EEG changed significantly,while scalp EEG did not change significantly. Only 9 patients had ischemic events in the corresponding supply territories after the occlusion in the cortical EEG significant change group. The changes in the depth of anesthesia had the consistent impact on cortical and scalp EEG. Conelusions Simultaneous monitoring of cortical and scalp EEG is a simple and effective method for monitoring cortical ischemia during anterior circulation intracranial aneurysm surgery, and may effectively identify the effect of anesthesia on EEG.

OBJECTIVETo probe the tendency of physiological and psychological change of armored vehicle drivers working continuously in hot environment.

METHODSTen cross-designed standard subjects were divided into control group and driver group. The driver group worked continuously in 32 degrees C approximately 38 degrees C and RH 50% approximately 80% environment. Heart rate, body temperature, systolic blood pressure and pulse pressure were synchronously and dynamically determined during work. Psychological parameters such as profile of mood states (POMS) questionnaire, serial addition/subtraction (SAS) and two-digit search (TDS) selected from neurobehavioral evaluation system (NES) before and after work were also tested.

RESULTSThe curves of the four main physiological parameters rapidly synchro-rose to a peak at (36 +/- 3) min, whereafter the curves descended smoothly. At (144 +/- 8) min, the curves climbed up to the second peak. The determination of the behavioral psychology revealed that the degree of fatigue of the drivers increased gradually when the working time prolonged, and the grip descended by great extent, excitement reduced markedly after work, but the negative mood, such as confusion and depression were increased, error of visual apperceive-operation was increased, right decreased, intelligence reduced, rapidity and delicacy of movement reduced.

CONCLUSIONThe physiological parameters of armored vehicle drivers working continuously in the hot condition appeared double-peak effect, the 1st peak was at (36 +/- 3) min, and the body was in mobilizing stage. Whereafter, the body was in smooth stage. And at (144 +/- 8) min, when the 2nd peak appeared, the body then was in physiological tolerance limit stage.

Adaptation, Physiological ; Adult ; Affect ; Automobile Driving ; psychology ; Body Temperature ; Fatigue ; epidemiology ; etiology ; Heart Rate ; Hot Temperature ; Humans ; Male

Objective: To observe the blood perfusion volume variation pattern in the body surface microcirculation at the Yuan-Primary and the Xi-Cleft points during the menstrual cycle in female college students with moderate constitution under normal physiological state of the uterus; to explore the specific laws of the body surface microcirculation at the Yuan-Primary and Xi-Cleft points in response to the uterine qi and blood changes under normal physiological conditions, and to provide the experimental basis for the specificity of acupoints reflecting the uterine function. Methods: Forty-three healthy and moderate constitution female college students with regular menstrual cycles, without dysmenorrhea and not yet giving birth were recruited. Bilateral Yuan-Primary points [Taichong (LR 3), Taibai (SP 3) and Taixi (KI 3)] and Xi-Cleft points [Zhongdu (LR 6), Diji (SP 8) and Shuiquan (KI 5)], belonging to the three yin meridians of foot and adjacent to the spinal cord segment of the uterus, were selected as the detection acupoints; the crossing point of the three yin meridians of foot [Sanyinjiao (SP 6)], the uterus-related meridian acupoint [Xuehai (SP 10)], the uterus-non-related meridian acupoint [Xuanzhong (GB 39)], and the non-meridian non-acupoint point were selected as the control points. The laser speckle blood flow imaging technique was used to monitor the blood perfusion volume in skin microcirculation at the above points at the menstrual, follicular, ovulatory, and luteal phases of the subjects. Results: The blood perfusion volume in the body surface microcirculation at the right Zhongdu (LR 6) at the ovulatory phase was higher than that at the menstrual, follicular and luteal phases (all P<0.05); there was no significant difference in the microcirculation blood perfusion volume at the other points among different phases (all P>0.05). Conclusion: The blood perfusion volume in the body surface microcirculation at Zhongdu (LR 6), the Xi-Cleft point of the Liver Meridian, shows a specific response to qi and blood changes in the uterus of women with moderate constitution.

BACKGROUND:For the replacement treatment of long-segment tracheal defects,although tissue engineering research has made some progress in recent years,it is still not perfect,and one of the biggest difficulties is that the hemodynamic reconstruction of the tracheal replacement cannot be achieved rapidly. OBJECTIVE:To preliminarily explore the potential of polycaprolactone scaffolds modified with exosome-loaded hydrogels to construct a rapidly vascularized tracheal substitute. METHODS:Exosomes were extracted from bone marrow mesenchymal stem cells of SD rats.After preparation of hyaluronic acid methacrylate solution,the exosome solution was mixed with hyaluronic acid methacrylate solution at a volume ratio of 1:1.Hyaluronic acid methacrylate hydrogels loaded with exosomes were prepared under ultraviolet irradiation for 5 minutes.The degradation of exosome-unloaded hydrogels and the controlled release of exosome-loaded hydrogels were detected.Polycaprolactone scaffolds were prepared by 3D printing.The pure hyaluronic acid methacrylate solution and the exosome-loaded hyaluronic acid methacrylate solution were respectively added to the surface of the scaffold.Hydrogel-modified scaffolds and exosome-modified scaffolds were obtained after ultraviolet irradiation.Thirty SD rats were randomly divided into three groups with 10 rats in each group and subcutaneously implanted with simple scaffolds,hydrogel-modified scaffolds and exosome-modified scaffolds,respectively.At 30 days after surgery,the scaffolds and surrounding tissues of each group were removed.Neovascularization was observed by hematoxylin-eosin staining and Masson staining and the expression of CD31 was detected by immunofluorescence. RESULTS AND CONCLUSION:(1)As time went by,the hydrogel degraded gradually,and the exosomes enclosed in the hydrogel were gradually released,which could be sustained for more than 30 days.The exosome release rate was faster than the degradation rate of the hydrogel itself,and nearly 20%of the exosomes were still not released after 30 days of soaking.(2)Under a scanning electron microscope,the surface of the simple polycaprolactone scaffold was rough.After hydrogel modification,a layer of gel was covered between the pores of the scaffold,and the scaffold surface became smooth and dense.(3)After 30 days of subcutaneous embedding,hematoxylin-eosin staining and Masson staining showed that more neovascularization was observed inside the scaffolds of the exosome-modified scaffold group compared with the hydrogel-modified scaffold group.The hydrogels on the scaffolds of the two groups were not completely degraded.Immunofluorescence staining showed that CD31 expression in the exosome-modified scaffold group was higher than that in the hydrogel-modified scaffold group(P<0.000 1).(4)These results indicate that hyaluronic acid methacrylate hydrogels can be used as controlled-release carriers for exosomes.The 3D-printed polycaprolactone scaffold modified by hyaluronic acid methacrylate hydrogel loaded with exosomes has good biocompatibility and has the potential to promote the formation of neovascularization.