BACKGROUND:Orthotopic lung transplantation model in a rat is the key to investigate the chronic rejection after lung transplantation. However, the precise surgical technique and difficult operation limit the application of the model. OBJECTIVE:To improve the process of anesthesia and lung transplantation, and to establish a rapid, safe and reversible rat lung transplantation model. METHODS:A total of 42 rats were used to establish the model, including 21 donor models and 21 receptor models. The donor lung was excised by median sternotomy with dissection of the left lung and implantation of cuffs (intravenous catheters cut into 1.5 mm sections). The left lung was implanted in the recipient by lateral thoracotomy using the cuffs for anastomoses. The duration of surgery and success rate of transplantation were recorded and calculated. RESULTS AND CONCLUSION:The survival rate of rats after lung transplantation was 100%. The time of left donor lung extraction was (35.3±5.1) minutes in average. The time of placing cuff in donor lung was (12.5±4.6) minutes in average. The surgical procedure time of recipient was (50.2±3.3) minutes. The time of arteriovenous and bronchus casing anastomosis was (27.7±6.2) minutes. After pulmonary artery and vein blood flow was disparked, the whole lung turned red rapidly, blood perfusion was sufficient, venous returned unimpeded;after mechanical ventilation resumed, al graft lungs expanded wel . This improved anesthesia and lung transplantation technique in rats can provide a valid, reliable and reproducible animal model for studying immune responses and rejection in lung transplantation.

AIM: To determine differentially expressed genes associated with cell adhesion and immune regulation in peripheral blood eosinophils from asthmatic patients. METHODS: Peripheral blood eosinophils were isolated from the asthmatic patients at the time of exacerbation and after improvement. Total RNA was extracted. Super SMART PCR cDNA was synthesized, suppression subtractive hybridization (SSH) and PCR-select differential screening technology were used to detect expressed genes. The differentially expressed genes were sequenced. RESULTS: High efficiency subtractive cDNA library was constructed successfully. Differential screening identified 15 differentially expressed genes, which were Charcot-Leyden crystal protein (CLC protein; galectin-10), putative pre-mRNA splicing regulator female-lethal (2D), aquaporin 9 (AQP9), IL-8, slingshot 2L (SSH-2L), PP1 catalytic subunit, beta isoform, helicase with zinc finger domain (HELZ), ?2-microglobin (?2-MG) and a gene associated with mitochondrion. CONCLUSION: Increased expression of these genes might be associated with eosinophil migration, adhesion and immune regulation. Intervention of these pathways may provide a theoretical base for future new targeting treatment for asthma.

Epithelial-Mesenchymal Transition ; Humans ; Liver Cirrhosis

Intervertebral disc degeneration is considered as a primary cause of clinical low back pain, however the molecular mechanism is not clear yet. Recently, researches on the molecular basis of intervertebral disc degeneration have become a hotspot. The special structure and biomechanics properties of the disc contribute to its propensity toward degeneration. Intervertebral disc degeneration is associated with the changes of the cytological behavior,including the increase in cell death and the degradation of extracellular matrix. However, the mechanism of cell death including cell apoptosis and autophagy in intervertebral disc degeneration remains unclear. Further study on the molecular mechanism of intervertebral disc degeneration is the foundation of improving and treating the intervertebral disc degeneration in the future. Although some progresses are made in the aspect of biological study, the biological environment of intervertebral disc itself is still a challenge for the development of biological treatment. This article is to review the latest advance on the biological characteristics of normal intervertebral disc and the cell death in the process of the intervertebral disc degeneration.
Animals ; Apoptosis ; Cell Death ; Extracellular Matrix ; metabolism ; Humans ; Intervertebral Disc ; cytology ; metabolism ; Intervertebral Disc Degeneration ; metabolism ; physiopathology

OBJECTIVETo compare the changes of inward rectifying K(+) (Kir) current (I(K1)) density in atrial myocytes of patients with rheumatic atrial fibrillation (RAF) less or longer than 6 months.

METHODI(K1) density was measured with whole-cell patch clamp technique in single myocyte isolated by an enzymatic dissociation method from right atrial appendages in patients with RAF less than 6 months (n = 18) and longer than 6 months (n = 18), RAF patients with normal sinus rhythm (NSR, n = 18) served as control.

RESULTSThe average resting membrane potentials were similar between various groups. I(K1) density in single myocyte at -50 to -100 mV of patients with RAF longer than 6 months was significantly increased compared to that in patients with RAF less than 6 months and NSR patients. I(K1) density in single myocyte at hyperpolarized potential level (-100 mV) was also significantly increased in patients with RAF longer than 6 months (8.94 +/- 0.15) than that in patients with RAF less than 6 months (4.35 +/- 0.49) and NSR patients compared to that in NSR (4.05 +/- 0.96, P < 0.05 vs RAF longer than 6 months).

CONCLUSIONThe data suggest I(K1) current increase might contribute to the electrical remodeling in RAF patients.

Adult ; Atrial Fibrillation ; metabolism ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Myocytes, Cardiac ; metabolism ; Patch-Clamp Techniques ; Potassium ; metabolism ; Potassium Channels, Inwardly Rectifying ; metabolism

Adult ; Female ; Humans ; Middle Aged ; Myxoma ; pathology ; surgery ; Vulvar Neoplasms ; pathology ; surgery

OBJECTIVETo observe the clinical significance of postoperative personalized antithrombotic therapy for patients with hemophilic arthritis (HA) patients after arthroplasty.

METHODSFrom September 2005 to October 2013, 11 cases of arthroplasty for hemophilic arthritis in hip and knee total operation 14 times,including 1 case of double knees (calculated as one operation), operation in left knees 6 times, operation in right knees 5 times, 2 in hip. All the patients were male and the age ranged from 23 to 57 years old,with an average of (36.1 ± 11.0) years old; the average weight was (64.1 ± 8.9) kg. All the patients were preoperatively diagnosed and classified as hemophilic arthritis with the radiological images and laboratory tests. According to the function of joints, the risk of postoperative venous thromboembolism (VTE), and dynamic observation of Factor VIII:C (FVIII:C) activity, patients were treated with personalized antithrombus by adjusting the dosage of recombinant human coagulation factor VIII (Kogenate FS). All the patients were orderly divided into postoperatively distal joints moving group and none-moving group to observe the coagulation function.

RESULTSThe enrolled patients had no postoperative complication of VTE and pulmonary embolism (PE). The APTT and D-2 were different between two groups in the postoperative early stage. Length of hospital day was shorter in the moving group than none-moving group.

CONCLUSIONBecause of the self-coagulation disorder, patients with HA tended to bleed. However it doesn't mean that there is no risk of postoperative thrombosis. Therefore,it's important to determine how to control the balance between postoperative antithrombus, hemostasis,and coagulation factor replacement therapy after arthroplasty for HA. Postoperative moving has proved helpful for HA, especially in reducing the risk of hemostasis and shortening the time in hospital.

Adult ; Arthritis ; surgery ; Arthroplasty ; adverse effects ; Factor XIII ; metabolism ; Hemophilia A ; complications ; Hemostasis ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Thrombosis ; prevention & control ; Young Adult

OBJECTIVETo investigate the effects of Shadu Cao Mixture (SDCM, traditional Chinese medicine) on immune functions of immunosuppression mice.

METHODSFifty BALB/C mice were randomly divided into blank control group, model group, SDCM low-dose, middle-dose and high-dose group. Except the blank control group, other groups were intraperitoneal injected with cyclophosphamide (40 mg/kg) to establish immunosuppression mice model. The blank control group and model group received gavage administration with nonnal saline, while the other groups received gavage administration with different doses of SDCM (10, 20, 40 m/kg for 15 days) respectively. The number of leukocytes and serum levels of interleukin-2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in peripheral blood, spleen index, and the function of NK cells were measured.

RESULTSCompared with the model group , SDCM increased the number of leukocytes and serum concentrations of IL-2, TNF-α and IFN-γ in peripheral blood and improved the spleen index and the function of NK cells significantly (P < 0.05-0.01).

CONCLUSIONSDCM could remarkably enhance the immune functions of immunosuppression mice induced by cyclophosphamide.

Animals ; Cyclophosphamide ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Immunosuppression ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Spleen ; immunology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo observe the antimicrobial activity of topical agents commonly used for burns on Acinetobacter baumannii (AB) in both free and biofilm states, and their synergistic effect with ambroxol on AB within biofilm.

METHODSEleven AB strains were isolated from wound excretion, respiratory tract, and blood of patients hospitalized in our hospital from August 2005 to April 2007. (1) The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of mafenide acetate and chlorhexidine acetate to free AB (including drug-resistant, drug-sensitive, and standard strains) were determined by dilution method. (2) AB was cultured with LB or TSB medium for 12, 24, and 48 h to form biofilm, and it was treated with above-mentioned two topical agents in MBC (mafenide group and chlorhexidine group) for 30 min. Biofilm not treated by topical agent was used as control group. The biofilm thickness was determined with confocal laser scanning microscope. The proportion of living bacteria in biofilm was calculated. AB biofilm in each topical agent group was mixed and inoculated into LB culture dish to observe the growth of bacteria. (3) AB was cultured with LB medium for 48 h to form biofilm, which was respectively treated by above-mentioned two topical agents in MBC (mafenide group and chlorhexidine group) and combination of each topical agent with 3.75 mg/mL ambroxol solution (ambroxol + mafenide group and ambroxol + chlorhexidine group) for 30 min. Biofilm not treated by topical agents was used as control group. Growth of bacteria in biofilm was detected with MTT method (denoted as absorbance value). Data were processed with one-way analysis of variance and LSD-t test.

RESULTS(1) MIC of mafenide acetate and chlorhexidine acetate for free AB was respectively 25.00 mg/mL and 0.03 mg/mL. MBC of both agents for free AB was the same as their MIC. (2) Among three groups, the thickness of biofilm of sensitive AB was thicker than that of drug-resistant bacteria at most of the time points. Compared with those in control group, biofilm thickness and proportion of living bacteria in biofilm were slightly decreased in mafenide and chlorhexidine groups. The growth of bacteria was abundant in each group. (3) Absorbance value of drug-resistant bacteria in control, mafenide, and chlorhexidine groups was respectively 0.776 ± 0.071, 0.625 ± 0.063, and 0.420 ± 0.068. Absorbance value of drug-resistant bacteria in ambroxol + mafenide group (0.174 ± 0.089) was significantly lower than that of control group (t = 11.823, P = 0.000) and mafenide group (t = 9.248, P < 0.01). Absorbance value of ambroxol + chlorhexidine group (0.178 ± 0.044) was significantly lower than that of control group (t = 16.009, P = 0.000) and chlorhexidine group (t = 6.681, P < 0.01).

CONCLUSIONSDrug-resistant AB forms biofilm readily, which prevents topical agents from killing the bacteria inside. Combined use of ambroxol with topical agents gives synergistic effect on killing AB in biofilm in the wound.

Acinetobacter baumannii ; drug effects ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; Biofilms ; drug effects ; Burns ; microbiology ; Chlorhexidine ; pharmacology ; Drug Resistance, Bacterial ; Humans ; Mafenide ; pharmacology ; Microbial Sensitivity Tests

AIMTo investigate the transformation of characteristics of epidermal cells from foreskin which were used to reconstruct male rabbit anterior urethra in combination with acellular collagen matrices.

METHODSIn three rabbits, autologous foreskin epidermal cells were isolated, expanded in vitro, and seeded (inoculated) onto a tubular acellular collagen matrix, acquired from allogeneic rabbit bladder submucosa. A urethral mucosal defect was created, and urethral reconstruction was performed with the tubular acellular collagen matrix seeded with epidermal cells.

RESULTSOn gross examination at 12 months following the procedure, the mucosa of the urethral grafts appeared lubricous and smooth. Urethrography showed that a wide urethral caliber had been maintained without any sign of strictures. Histological examination showed a transitional cell layer in the graft without evidence of a margin between the graft and the host tissue at 12 months postoperatively.

CONCLUSIONEpidermal cells seeded onto acellular collagen matrices can be successfully used to reconstruct urethras that have defects and are transformed to transitional epithelial cells.

Animals ; Cell Transplantation ; methods ; Collagen ; Epidermis ; cytology ; Foreskin ; cytology ; Graft Survival ; Male ; Mucous Membrane ; cytology ; Rabbits ; Reconstructive Surgical Procedures ; methods ; Tissue Culture Techniques ; Tissue Engineering ; methods ; Urethra ; surgery ; Urethral Stricture ; surgery