Objective To explore an easier method to get epithelial cell and fibroblast from a rabbit' s trachea for trachea tissue engineering in vitro. Methods Digest the epithelial cell and fibroblast by trypsin after co-culture for 6 days. Distinguish these two kinds of cells according to their different tolerances to trypsin and culture characteristics. The new method and the traditional method were compared by the proliferation of the cells with cell counting and MIT assay. Results The purity of the cells got with this new method is up to 100% . The cells proliferation is as same as that cultured by traditional methods. Conclusion With this method, one can get two kinds of different cells from the original in one time, it is easier to practice than the traditional.

OBJECTIVE To study the antibiotic sensitivities and characterization of ?-lacamase activity and the antibiotic resistance mechanisms.METHODS The extended-spectrum ?-lactamases were detected.Agar dilution method was used to determine the minimum inhibitory concentration(MIC)of severel antibiotics.The plasmids of clinical isolates were cloned into competent cells(K-12).RESULTS Isoelectric focusing(IEF):The last 5 clinical isolates have all got an activity at a high pH 9 which could be presumptively identified as AmpC.The last two clinical isolates had a weak activity with pH of about 8.0,which belonged to SHV type ?-lactamases.All the isolates had a group of activities at the lower pH range of the gel(pH5).These could be presumptively identified as TEM ?-lactamases.CONCLUSIONS The data suggest that each isolate clearly originate from the same population but possess mutations that leads to antibiotic resistance.The combinations of mutations in same isolates suggest that an earlier strain have infected other compartments and become the founder for later antibiotic resistant variants.

Objective To investigate the MRI findings of sinonasal inverted papilloma so as to promote the diagnostic accuracy.Methods All 36 cases of sinonasal inverted papilloma were verified by pathology.Their MRI findings were analyzed retrospectively.Results The lesion occurred in the lateral nasal wall in 25 cases,in the maxillary sinus in 4 cases,in the ethmoid sinus in 2 cases,in the frontal and ethmoid sinuses in 2 cases, in the ethmoid and sphenoid sinuses in 2 cases and in the frontal sinus in one case.Of the 36 cases,11 cases had recurrent lesions and 3 cases were associated with malignant change.All lesions showed well-defined margin,with lobulate configuration in 26 cases and irregular shape in 10 cases.The diameter of the lesions ranged from 22 to 82 mm (mean,38 mm).On MR T1WI,sinonasal inverted papilloma revealed isointense signal compared to adjacent muscle in 32 cases and slight hyperintense signal in 4.On T2 WI,the lesions showed heterogeneous hyperintense signal in 34 cases and inhomogeneous isointense signal in 2 cases with malignant change.Postcontrast MR imaging demonstrated marked inhomogeneous enhancement compared with nasal septum mucosa in 34 cases.Regular fence-like pattern was found on MR T2WI and enhanced T1WI in 32 cases,while irregular ones were found in three cases with malignant change.The time-intensity curve (TIC)of dynamic contrast enhancement of MRI showed rapid enhancing and slow wash-out type in 7 cases,while rapid enhancing and wash-out pattern was detected in one ease with malignant change.Conclusion Regular fence-like pattern on MR T2WI or enhanced T1WI was reliable characteristics for the diagnosis of sinonasal inverted papilloma.Irregular ones may suggest malignant change.

OBJECTIVETo explore the rapid neutrophil engraftment and long-term hematopoietic reconstitution.

METHODSMononuclear cells (MNCs) were isolated from 5-Fu treated male BDF1 mouse bone marrow and CD(34)(+)/c-kit(+) cells were selected from the MNCs by using MoFlo Cell Sorter. MNCs and CD(34)(+)/c-kit(+) cells were co-cultured with mouse bone marrow-derived mesenchymal stem cells (MSCs) in a two-step expansion. The expanded cells were then transplanted into sublethally irradiated female BDF1 mice.

RESULTSCo-culture with MSCs resulted in 10.8-, 4.8-, 65.9- and 38.8-fold increases yields of median total nucleated cells, CD(34)(+) cells, GM-CFC and HPP-CFC, respectively, as for the MNCs culture, and 76.1-, 2.9-, 71.7- and 51.8-fold increase respectively for the CD(34)(+)/c-kit(+) cell culture. The expanded cells could rapidly engraft in the sublethally, irradiated mice, reconstitute their hematopoiesis, and be detected in the recipients bone marrow 2 months after transplantation.

CONCLUSIONSHematopoietic stem/progenitor cells co-cultures with MSCs in two-step expansion could increase expansion yields of total nucleated cells, GM-CFC and HPP-CFC. The availability of increased numbers of expanded cells may result in more rapid engraftment of neutrophils following infusion to transplant recipients.

Animals ; Antigens, CD34 ; analysis ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; physiology ; Male ; Mice

OBJECTIVETo analyze the long-term limb sparing outcomes of various reconstructive procedures and complications associated with extensive excision in bone tumours of the shoulder girdle.

METHODSShoulder reconstruction following resection of bone tumors of the shoulder girdle was reviewed retrospectively for 32 patients at the Wesley Hospital or Princess Alexandra Hospital. The reconstructions were including arthrodesis (8 cases), allograft-prosthetic composite (7 cases), spacer (6 cases), no bone reconstruction (5 cases), prosthesis (3 cases), vascular fibular graft (2 cases) and allograft (1 case).

RESULTSThe average duration of follow-up was 81 months for the 23 patients who were still alive at the time of the latest follow-up examination. Functional results were related to the type of resection and the method of shoulder reconstruction. After intra-articular resection of the proximal humerus with loss of the abductor mechanism, arthrodesis resulted in 87% functional score and more strength was found after reconstruction with prosthesis or allograft-prosthetic composite. Allograft-prosthetic composite had better function (Score 79%) than prosthesis alone after intra-articular resection of the humerus because reconstruction of the deltoid and the rotator cuff could be performed without increased prevalence of complication related to the allograft. After extra-articular resection of the glenoid cavity and the proximal humerus with abductor mechanism, reconstruction with a functional spacer frequently resulted in superior subluxation of the implant and only fair function (Score 66%) of the shoulder. With two teen-aged patients, a free fibular graft inserted after intra-articular resection of the proximal humerus led to fair function (Score 73%), to be followed by secondary arthrodesis when growth was complete.

CONCLUSIONSIndications for the method of reconstruction depend on type of resection, age, gender, occupation, the expected functional level and restriction of activity. After resection of the abductor mechanism, arthrodesis results in more strength and is the preferred option for the young active demanding patients. If the abductor mechanism is preserved, allograft-prosthetic composite gives good results.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; surgery ; Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Middle Aged ; Orthopedic Procedures ; methods ; Osteosarcoma ; surgery ; Retrospective Studies ; Shoulder Joint ; surgery ; Treatment Outcome

OBJECTIVETo explore the biological characteristics of mesenchymal stem cells (MSC) derived from umbilical cord blood (UCB) and their supporting capacities in ex vivo expansion of hematopoietic stem/progenitor cells (HSPCs).

METHODSLow-density mononuclear cells (MNCs) from UCB were cultured in IMDM containing 20% FBS to form confluent adherent cells through 15 passages. Some cytokines in the conditioned medium were determined with ELISA. UCB-derived adherent cells were displayed with antibodies and analyzed with flow cytometry. The supporting capacity of UCB-derived adherent cells for ex vivo expansion of CD34(+) cells was assayed by co-culture in a two step culture. UCB-derived adherent cells were induced for chondrogenic differentiation with chondrogenic medium, and the induced cells were analyzed for the type II pro-collagen gene expression with RT-PCR.

RESULTSThe mean number of adherent fibroblast like colonies derived from UCB was (3.5 +/- 0.7)/10(6) MNCs. UCB-derived MSCs could survive for at least 15 passages of expansion. In their undifferentiated status, UCB-derived MSCs were CD13(+), CD29(+), CD90(+), CD105(+), CD166(+), SH2(+), SH3(+), SH4(+), CD45(-), CD34(-), and CD14(-). Stem cell factor (SCF), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) could be detected in the supernatant of the cultures. The MSCs cultured in chondrogenic media could differentiate into chondrogenic cells and express type II pro-collagen mRNA. UCB-derived MSCs could support the proliferation and differentiation of UCB CD34(+) cells in vitro.

CONCLUSIONUCB-derived MSCs are similar to those derived from adult bone marrow and can support the proliferation of hematopoietic stem/progenitor cells.

Antigens, CD34 ; metabolism ; Cell Adhesion ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Collagen Type II ; genetics ; Cytokines ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo characterize the different varieties of Pseudostellaria heterophylla during cultivation.

METHODUsing systematic selection in the main productive areas, the techniques of random design, all varieties were observed for 3 years.

RESULTThe biological and 425 productive characteristics of P. heterophylla var. macrophylla, P. heterophylla var. Foliolum, and P. heterophylla var. anvense were significantly different (P < 0.01). There were also differences in ecological adaptability, plant characteristics, pollen granule, chromosomes, and isoenzyme of the three cultivars.

CONCLUSIONThe strain types of P. heterophylla was denominated for the first time. The characteristics and productivity index system of P. heterophylla varieties were determined.

Caryophyllaceae ; anatomy & histology ; enzymology ; genetics ; Catechol Oxidase ; analysis ; Chromosomes, Plant ; Ecosystem ; Flowers ; anatomy & histology ; Lipase ; analysis ; Peroxidase ; analysis ; Plant Leaves ; anatomy & histology ; Plant Roots ; anatomy & histology ; Plants, Medicinal ; anatomy & histology ; enzymology ; genetics

INTRODUCTIONDeveloping effective exercise programmes for the paediatric population is a strategy for decreasing obesity and is expected to help in eventually limiting obesity-associated long-term health and societal impact. In this study, the effects of a 12-week twice weekly additional exercise training, which comprised a combination of circuit-based resistance training and aerobic exercises, in additional to typical physical education sessions, on aerobic fitness, body composition and serum C-reactive protein (CRP) and lipids were analysed in 13- to 14-year-old obese boys contrasted with a control group.

MATERIALS AND METHODSBoth the exercise group (EG, n = 12) and control group (CG, n = 12) participated in the typical 2 sessions of 40-minute physical education (PE) per week in schools, but only EG participated in additional 2 sessions per week of 45 to 60 minutes per session of exercise training, which comprised a combination of circuit-based resistance training and aerobic exercises maintained at 65% to 85% maximum heart rate (HRmax = 220 - age). Body composition was measured using dual energy X-ray absorptiometry (DEXA). Fasting serum CRP and blood lipids were analysed pre- and postexercise programme. Aerobic fitness was measured by an objective laboratory submaximal exercise test, PWC170 (Predicted Work Capacity at HR 170 bpm).

RESULTSExercise training significantly improved lean muscle mass, body mass index, fitness, resting HR, systolic blood pressure and triglycerides in EG. Serum CRP concentrations were elevated at baseline in both groups, but training did not result in a change in CRP levels. In the CG, body weight increased significantly at the end of the 12-week period.

CONCLUSIONThis study supports the value of an additional exercise training programme, beyond the typical twice weekly physical education classes, to produce physiological benefits in the management of obesity in adolescents, including prevention of weight gain.

Adolescent ; C-Reactive Protein ; analysis ; metabolism ; Exercise Therapy ; Humans ; Lipids ; analysis ; blood ; Male ; Obesity ; physiopathology ; therapy ; Physical Fitness ; physiology

Aim To investigate the eorrelation between angiotensin II (Ang II ) level and clinical indicators in patients with rheumatoid arthritis ( HA) , and to determine the therapeutic effect of angiotensin receptor blockers ( ARBs).Methods Plasma samples and personal information were collected from HA patients admitted to our hospital from 2019 to 2021.The level of Ang II in plasma was determined by ELISA to elucidate the correlation between plasma Ang II level and the severity of HA.The pathological changes of synovi-al tissues and T eells subtype in different groups of HA patients were determined by pathological examination and flow cytometry.A rat model of collagen-induced arthritis (CIA) was established and the pathological examination was used to confirm that valsartan could alleviate the disease course in the CIA animal model.Results Compared with control group, the plasma level of Ang II in HA patients significantly increased.After therapy with oral ARBs plasma Ang H levels and anti - cyclic citrullinated peptide antibody ( CCP) titre were significantly lower than those untreated HA patients.The level of Ang II in plasma was positively correlated with CCP and the number of monocytes, but negatively with number of RBC and hemoglobin content.Staining of synovial tissue with HE and Masson found that patients with HA had significant synovial proliferation, pannus formation , and numerous inflammatory cell infiltrates compared with control patients.Immunohistochemical results showed significant infiltration of CD4 4 T cells in synovial tissues of HA patients.Western blot and immunofluorescence analysis showed that the expression of angiotensin type 1 receptor ( ATI R ) was significantly up-regulated in CD4 + T cells and synovial tissues of HA patients.The results of animal experiments showed that valsartan harl therapeutic effect on CIA rats and could delay the disease process of CIA.Conclusions Plasma Ang II level is positively correlated with CCP level and HA severity.ARBs can down-regualte CCP level and delay disease progression in HA patients.Animal experiments showed that valsartan blocks the combination of Ang H and ATI R and has therapeutic effect on a CIA rat model.This study provides the theoretical and experimental basis for ARBs to become the preferred antihypertensive drugs for HA patients with hypertension.

Aim To explore the important role of HDAC5 in P-gp expression in rats in high-altitude low oxygen environment and its effect on phenytoin sodium pharmacokinetics. Methods Wistar rats were transported to Batang, Yushu, Qinghai, at an altitude of 4010 m, with 6 rats in each group, divided into 1 d and 3 d groups. Different groups were given phenytoin, phenytoin combined with hypericin, and phenytoin combined with verapamil. Plasma and liver tissues were collected at different time after taking the drug in the plateau area. The concentration of phenytoin sodium in plasma was determined by UFLC-MS method. Changes in protein expression were detected by Western blot. Results The results of UFLC-MS showed that the AUC