BACKGROUND: The problem to be solved firstly in adipocyte implantatior after its in vitro adherent ulture f preadipocyte with scaffold using tissue-engineered technique is the biocompatibility of scaffold and dipocyte.OBJECTIVE: To investigate the feasibility of poly-β-hydroxybutyric acid (PHB) used for readipocyte implantation.DESIGN: Randomized controlled trial.SETTING: Medical College of Wuhan University.MATERIALS: This trial was carried out in Medical College, Wuhan University during October 2003 o June 2005. Twelve male SD rats(Experimental Animal Center, Medical College, Wuhan University), weighing rom 200 to 250 g, were involved.PHB scaffold was provided by Institute of Polymer Materials, Department f hemical Engineering, Tsinghua University.METHODS: The preadipocytes of rats were isolated, purified and ultured in vitro for use. PHB scaffold was made into 0.75 cm×0.75 cm ×0.2 cm lamellar cell biological caffold, then which were soaked by DMEM/F12 culture medium containing 150 g/L fetal bovine serum and laced in culture plate, one piece each well, 12 wells totally. 1.0 mL cultured cell suspension was added n each well to prepare cell scaffold complex. Eight rats were selected. The prepared complexes were implanted into right-side back tissue of a rat and fixed with 5-0 silk, serving as adipocyte scaffold roup; the biological scaffolds without adipocyte adherence were implanted into left-side back tissue of he same rat with the same method, serving as blank scaffold group. One lamellar biological scaffold was mplanted into each side. Eight weeks later, the appearance and structural change of grafts were observed, nd the grafts were weighted and their volumes were measured. Four lamellar biological scaffolds were mplanted into the other 4 rats separately and taken out after 3 weeks, then they were fixed by 100 g/L eutral formalin, sliced, and stained by haematoxylin and eosin (HE). Their histological changes were observed.MAIN OUTCOME MEASURES: ① Gross and histological observation of grafts in adipocyte scaffold roup and blank scaffold group. ② Comparison of volume and mass of grafts in two groups.RESULTS: Twelve ets were involved in the result analysis ,without deletion. ①Gross observation: In the adipocyte scaffold group, grafts presented flesh-color appearance and peplos in the peripheral region, and newly ormed minute blood vessels ingrowing into the scaffold. In the blank scaffold group, grafts presented rey ppearance, which was coated by peplos. ② HE staining: Three weeks later, a few scattered adipocytes were ound in the peripheral region of the grafts,there were very few vessels. Eight weeks later, lamellar ibrous peplos was found around the complex. Adipocytes ingrew from the peripheral region of scaffold into caffold, partial scaffold was nearly full of adipocytes and many vessels formed;In the blank scaffold roup, only peplos and vessels were found, scaffold was full of fibrous connective tissue and adipocytes ere not found. ③ Comparison of volume and mass: adipocyte scaffold group was superior to blank group in scaffold volume and mass [(257.5±70.2)vs.(144.6±62.6)mm3,(245.6±58.2) vs.(148.7±60.3)mg, both P＜0.01].CONCLUSION: Preadipocyte can adhere to PHB scaffold, proliferate and differentiate into mature dipocytes. It is feasible for PHB material to serve as a carrier for preadipocyte.

Objective To observe the effect of different doses of sufentanil on intubation guided by fiber bronchoscope in diffi‐cult airways .Methods One hundred and eighteen patients with difficult airways who underwent tracheal intubation were randomly divided into 3 groups [groupⅠ(n=40) ,groupⅡ (n=42) ,and groupⅢ (n=36)] .First all group received 1 μg/kg dexmedetomi‐dine ,then groupⅠ ,groupⅡ and group Ⅲ received sufentanil 0 .1μg/kg ,0 .2 μg/kg ,0 .3μg/kg infusion in bolus respectively .Heart rate(HR) ,mean arterial pressure(MAP)and saturation of pulse oximetry(SpO2 )were recorded at 5 min after patients arriving at operation room (T0 ) ,after drug infusion(T1 ) ,the time of intubation and seeing the epiglottis (T2 ) ,1 min(T3 )and 5 min(T4 )after intubation .side effect was recorded throughout the process .Results HR at T2 and T3 in groupⅡ and group Ⅲ were significantly lower than those in group Ⅰ(P<0 .05) .MAP at T2 and T3 in groupⅡ and group Ⅲ were significantly lower than those in groupⅠ(P<0 .05) .SpO2 at T2 in group Ⅲ was significantly lower than those of groupⅠ and groupⅡ(P<0 .05) .Cough reflex in groupⅠwas much frequent than others .Only group Ⅲ had respiratory inhibition .Conclusion Intravenous sufentanil (0 .2 μg/kg)could in‐hibit effectively stress reaction from endotracheal intubation ,in which less side effects such as haemodynamic changes and respirato‐ry depression occur .

Background and purpose:MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many tumors. This study investigated which miRNA might negatively regulate the expression of Aurora-B in osteosarcoma cells, and to lay the foundation for the further investigation of the effort and regulation of Aurora-B in osteosarcoma malignant phenotype.Methods:Bioinformatics prediction software (http://www.targetscan.org) and luciferase assays were used to investigate which miRNA might target to modulate the Aurora-B. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were used to further verify which miRNA could negative regulate the expression ofAurora-B gene.Results:Bioinformatics prediction showed let-7 family have the possibility to modulate the expression of Aurora-B; Luciferase assays showed thatAurora-B might be the target gene of let-7a/b/c/d/e/f/g/i; RTFQ-PCR and Western blot analysis testiifed that both the expression levels of Aurora-B mRNA and Aurora-B protein were signiifcantly decreased in Let-7a/g/i up-regulated U2-OS and HOS cells, compared to the cells in the negative control group; but in Let-7b/c/d/e/f up-regulated U2-OS and HOS cells, the expression levels of Aurora-B mRNA and Aurora-B protein have no signiifcant difference, compared to the cells in the negative control group.Conclusion:Let-7a/g/i may downregulate the expression of Aurora-B in human osteosarcoma cells.

Objective High expression of the valosin-containing protein ( VCP) gene can enhance the metastasis of osteosar-coma via the AKT/PI3K/NF-KappaB/MMP-9 signaling pathway, but the molecular mechanisms underlying the up-regulation of VCP in osteosarcoma cells remains unknown .This study aimed to determine whether miRNA-129-5p can regulate the VCP expression and its targets in human osteosarcoma cells . Methods The microRNA target-predicting software TargetScanhuman 6.2 ( http://www.tar-getscan.org/) was used to predict the possible targets of miRNA-129-5p on the VCP gene.Then, two recombinant gene report vectors containing the wild VCP gene 3′UTR ( psi-VCP vector ) and mutant VCP gene 3′UTR ( psi-VCPmut vector ) were constructed , se-quenced, and identified.The human osteosarcoma U2-OS cells were co-transfected with miRNA-129-5p mimic and psi-VCP vector or psi-VCPmut vector, respectively.A non-specificity mimic transfection served as negative control , and the luciferase activity was detec-ted in each group. Results The software prediction showed only one conserved function site of miRNA-129-5p on the VCP gene 3′UTR163-169 bp.Luciferase activity was significantly lower in the psi-VCP vector +miRNA-129-5p transfection group (15.529 ± 1.902) than in the VCP control group (21.781 ±0.854), VCP mutation experimental group (19.978 ±1.377), and VCP mutation control group (21.952 ±1.516) (P<0.05), with no remarkable difference between the VCP mutation control and VCP control groups (P=0.276). Conclusion miRNA-129-5p can probably regulate the targets of the VCP gene in human osteosarcoma U 2-OS cells.

Objective To compare the effectiveness,correlativity,and acceptability of the three methods:bone marrow smear,biopsy and flow cytometry in diagnosis of lymphoma patients with bone marrow involvement.Methods 68 cases of early stage lymphoma were studied by observing and comparing positive rates of the three methods:bone marrow smear,biopsy and flow cytometry.Results After confirming the bone marrow involvement,the positive rates for the 68 cases using bone marrow smear,biopsy and flow cytometry were 16.2 ％ (11/68),33.8 ％ (23/68) and 10.3 ％ (7/68) respectively.Bone marrow biopsy had the highest positive rate compared to the other methods.It was statistically significant when comparing the differences of the 3 methods (P ＜ 0.05).According to the correlativity analysis of the 3 methods,bone marrow smear and biopsy correlated with each other (P =0.002),while flow cytometery did not (P =0.270).Conclusions Morphological examination of the bone marrow smear is a fundamental method to test bone marrow involvement in lymphoma.Bone marrow biopsy creates the highest positive rate and has clear advantage compared to the other methods,however,bone marrow smear must be used at the same time as a complement.

OBJECTIVETo investigate the role of transforming growth factor β1 (TGF-β1) in patients with colorectal cancer.

METHODSFresh peripheral blood were obtained from 50 patients (before surgery and at least one week after surgery) and 25 healthy donors in the morning. Fresh colorectal cancer tissues and the adjacent tissues (at least 5 cm from the tumor site) were obtained from patients undergoing tumor resection. The expression levels of TGF-β1 in the blood and tissue specimens were determined using ELISA.

RESULTSThe plasma levels of TGF-β1 in patients with colorectal cancer were significantly higher than those in the healthy donors, and decreased after the surgery (P<0.05). The tumor tissues expressed higher levels of TGF-β1 than the adjacent tissues from both CEA-negative and -positive patients. The plasma level of TGF-β1 in the patients were positively correlated with the tumor size and clinical tumor stage (P<0.05).

CONCLUSIONTGF-β1 combined with CEA can provide important information for the diagnosis, prognostic assessment and prediction of recurrence in patients with colorectal cancer, and may provide new insights for anti-TGF-β1-based tumor immune therapeutic strategies.

Biomarkers, Tumor ; metabolism ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; metabolism ; Humans ; Neoplasm Recurrence, Local ; Prognosis ; Transforming Growth Factor beta1 ; metabolism

Objective:To investigate the clinical characteristics of elderly patients with coronavirus disease 2019(COVID-19), in order to provide scientific evidence for the diagnosis and treatment of COVID-19 in elderly patients.Methods:Clinical data of 102 patients with COVID-19 admitted to the B11 East Ward of the Zhongfaxincheng campus and the E1-3 ward of the Guanggu Campus of Tongji Hospital affiliated to Huazhong University of Science and Technology in Wuhan from 1 February 2020 to 28 February 2020 were retrospectively collected and analyzed.Patients were categorized into 2 groups: the elderly group(≥60 years old)and the young and middle-aged group(<60 years old). Differences in epidemiological features, demographics, clinical symptoms, laboratory results and imaging findings between the two groups were retrospectively analyzed.Results:Among 102 patients with COVID-19, 58 were in the elderly group(≥60 years old), with a median age of 67.0(63.8, 71.0)years old, and 44 in the young and middle-aged group(<60 years old), with a median age of 47.5(38.0, 51.8)years old.There was no significant difference in gender ratio between the two groups( χ2=0.033, P=0.855). Of 102 patients, 42.0%(21/50)had close contact with an infected person, 14.0%(7/50)were from infection clusters, and 18.0%(9/50)had suspected hospital-acquired infections.Fever and cough remained the most common symptoms, but gastrointestinal symptoms such as nausea, poor appetite, diarrhea and muscle cramps were also warning signs.Fatigue and cough were the most common presenting symptoms in elderly male patients.Bilateral patchy infiltrates(57.9%, 22/38)and ground-glass opacities(42.1%, 16/38)were the main imaging features and 42.1%(16/38)patients had multiple areas of the lungs involved.Over 50% patients had increased levels of blood glucose, D-dimer, fibrinogen, C-reactive protein, procalcitonin, multiple cytokines and neutrophil-to-lymphocyte ratio, as well as decreased levels of albumin, hemoglobin, hematocrit, lymphocytes and serum calcium.Compared with the young and middle-aged group, the elderly group had higher rates of abnormality in levels of D-dimer and serum calcium( χ2=7.067 and 4.166, P=0.008 and 0.041). Conclusions:Fever and cough are the most common symptoms in elderly patients with COVID-19.Elderly patients with COVID-19 have multiple abnormalities in clinical laboratory test results, which show a certain level of specificity compared with young and middle-aged patients.

To investigate the effects of Mycobacterium tuberculosis heat shock protein 65(HSP65)on Treg/Th17 immune balance in ApoE-knockout(ApoE-/-)mice, ApoE-/- mice with a high-cholesterol diet were immunized with M. tuberculosis HSP65. Sera were obtained for measurement of anti-HSP65 antibodies by ELISA; the effect of administration of different antigens was investigated, respectively, using flow cytometry analysis on the number of CD4+CD25+Foxp3+Tregs and CD4+IL-17+ Th17; the production of cytokines(IL-10, TGF-β1, IL-17 and IL-21)by these cells were determined by ELISA; total plasma cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were detected by biochemical autoanalyzer. Atherosclerotic lesions were measured by lipid deposition stained with oil red O. The results demonstrated that the levels of anti-HSP65 IgG antibodies were increased significantly in Mycobacterium tuberculosis HSP65-treated ApoE-/- mice, revealed obvious decrease in Treg number, Treg related cytokines(IL-10, TGF-β1)levels and significant increase in Th17 number, Th17 related cytokines(IL-17 and IL-21)levels, the levels of TC, TG, HDL-C and LDL-C did not change between groups, while the atherosclerotic lesions significantly increased. Results indicate that M. tuberculosis HSP65 could interrupt the Th17/Treg immune balance in ApoE-/- mice, suggesting a potential role in the formation and progression of atherosclerosis.

Objective To investigate the biomarkers for Parkinson disease(PD) by analyzing differentially expressed proteins in the serum of patients with PD. Methods A total of 24 patients with PD who were diagnosed in Beijing Hospital were enrolled,among whom 12 patients only had a medical history of PD(group P1) and the other 12 had underlying diseases such as hypertension and diabetes(group P2). A total of 24 individuals,matched for age and sex,who underwent physical examination were enrolled as control groups C1 and C2.Serum samples were collected,and after high-abundant proteins were removed,the label-free quantitative proteomics technique was used to measure the expression of proteins in serum. The double comparison method was used for comparison between groups P1 and C1 and between groups P2 and C2 to screen for differentially expressed proteins,and the proteins with a consistent changing trend(upregulation or downregulation) were identified as the differentially expressed proteins for PD,which were analyzed and interpreted by bioinformatics methods. Results Comparison between groups identified four differentially expressed proteins,i.e. PRG4,CFHR-3,ACTG1,and HIST2H2BF,among which PRG4 and CFHR-3 showed upregulated expression and ACTG1 and HIST2H2BF showed downregulated expression in the serum of PD patients,and a certain degree of interaction was observed. Conclusion Label-free quantitative proteomics can be used to identify the differentially expressed proteins in PD,which may have a certain value in the diagnosis of PD.

Objective To investigate the role of transforming growth factor β1 (TGF-β1) in patients with colorectal cancer. Methods Fresh peripheral blood were obtained from 50 patients (before surgery and at least one week after surgery) and 25 healthy donors in the morning. Fresh colorectal cancer tissues and the adjacent tissues (at least 5 cm from the tumor site) were obtained from patients undergoing tumor resection. The expression levels of TGF-β1 in the blood and tissue specimens were determined using ELISA. Results The plasma levels of TGF-β1 in patients with colorectal cancer were significantly higher than those in the healthy donors, and decreased after the surgery (P<0.05). The tumor tissues expressed higher levels of TGF-β1 than the adjacent tissues from both CEA-negative and-positive patients. The plasma level of TGF-β1 in the patients were positively correlated with the tumor size and clinical tumor stage (P<0.05). Conclusion TGF-β1 combined with CEA can provide important information for the diagnosis, prognostic assessment and prediction of recurrence in patients with colorectal cancer, and may provide new insights for anti-TGF-β1-based tumor immune therapeutic strategies.