[Objective]To investigate the rlpairing effect on articular cartilage defects by composite of cocultures of autogenic bone marrow-derived mesenchymal stem cells(BMSCs) and chondrocytes with allogenic fully deproteinized bone(FDB),in order to provide basis for optimizing seeding cells resources.[Method]Seeding cells were collected from two-passaged BMSCsand chondroeytes and then cocultured at the rate of 2 to 1.Full thickness articular cartilage defects in the knee joints of rabbits repaired by cocultured cells seeded into allogenic FDB were served as experimental group A,by simple FDB as control group B and by nothing as blank control group C.Repaired tissues were evaluated with macroscopic views,histological scores and immunohistochemistrical stains at 8 and 16 weeks postoperatively.[Result]Chondrocytes cocultured riched in extracellular matrix and proliferated promptly.In A regenerated tissues represented hyaline-like,smoothness and flat.In group B and C,repaired tissues were fiberous and no repaire in group C.Histological scores of experimental group A excelled group B and C with statistically significant differences(P0.05).Immunohistochemistrical stains showed that cells in the zones of repaired tissues were larger in size,arranged columnnedly,riched in type-Ⅱ collagen matrix and integrated satisfactorily with native adjacent cartilages and subchondral bones in the experimental group A at 16 weeks postoperatively.[Conclusion]Cocultures of autogenic BMSCS with chondrocytes can promote proliferation of chondrocytes and production of chondral matrix.Cocultures as seeding cells can save a number of chondrocytes,shorten culturing periods and reduce subcultured times.Cocultures embedded into FDB can repair articular cartilage defects effectively.

OBJECTIVE:To investigate synergistic effect of carbapenems combined with fosfomycin(FOS)on carbapenems-re-sistant Pseudomonas aeruginosa isolates from urinary tract infections in vitro. METHODS:The minimum inhibitory concentration was detected using agar double dilution method. The fractional inhibitory concentration index was determined by checkerboard meth-od. The effect of carbapenems combined with FOS on biofilm of P. aeruginosa isolates was determined using 96 crystal violet stain-ing. RESULTS:12 strains of carbapenem-resistant P. aeruginosa isolates were highly sensitive to FOS and amikacin,and were com-pletely resistant to imipenem and meropenem. The combination of imipenem with FOS could induce a synergistic effect on 4 strains (33.3%);meropenem combined with FOS could induce a synergistic effect on 5 strains(41.7%);no antagonistic effect of carbap-enems combined with FOS appeared. FOS combined with carbapenems could inhibit the biofilm of carbapenems-resistant P. aerugi-nosa(P<0.05 or P<0.01). CONCLUSIONS:The combination of carbapenems with FOS possesses in vitro synergistic antibacteri-al effect on part of carbapenems-resistant P. aeruginosa isolates,the mechanism of which may be associated with inhibiting the bio-film.

AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet.
METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods.
RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the
model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon.
CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.

Objective To study the biofilm formation ability and related gene distribution of Staphylococcus (S .) aureus iso‐lated from urinary tract infections to provide the theoretical basis for the prevention and treatment of clinical infection .Methods The minimal inhibitory concentration was detected using the agar double dilution method .The bacterial adhesion ability was deter‐mined by flat colony counting method .The biofilm formation ability was analyzed by the 96‐well crystal violet staining method .The biofilm‐associated genes were detected by PCR amplification .Results Eleven clinical strains of S .aureus were high resistant to pen‐icillin and erythromycin ,whereas were all sensitive to vancomycin and nitrofurantoin .All the isolates had a strong ability of adhe‐sion ,but the biofilm formation ability was weak .Among them ,the icaAD and icaBC genes were amplified in 10 S .aureus isolates . Conclusion The adhesion ability and biofilm formation ability of S .aureus isolated from urinary tract infections have the strain differences ,and ica is an important gene of S .aureus biofilm formation .

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics

OBJECTIVETo observe the clinical efficacy of comprehensive Western medical treatment plus Compound Yindan Decoction (CYD) in treatment of acute cholestatic hepatitis (ACH).

METHODSUsing randomized controlled study, 60 ACH patients in line with inclusive criteria were randomly assigned to the treatment group (treated by comprehensive Western medical treatment plus CYD) and the control group (treated by comprehensive Western medical treatment alone), 30 in each group. Scores for symptoms and levels of liver functions [total bilirubin (TBIL), direct bilirubin (DBIL), alkaline phosphatase (ALP), glutamyl transpeptidase (GGT), total biliary acid (TBA)] were observed before and after treatment.

RESULTSCompared with before treatment in the same group, total scores for symptoms decreased in the treatment group and the control group at the end of the 1st and the 4th week after treatment (all P < 0.05). Compared with the control group, total scores for symptoms decreased in the treatment group at the end of the 1st week (P < 0.05). Compared with before treatment, serum levels of TBIL, DBIL, ALP, GGT, and TBA all decreased in the two groups at the end of the 4th week after treat- ment (P < 0.01). Compared with the control group, serum levels of TBIL, DBIL, ALP, GGT, and TBA all decreased in the treatment group at the end of the 1st and the 2nd week after treatment (P < 0.05). Compared with the control group, the average time for TBIL and DBIL decreasing to the level less than five times the normal value was significantly shorter in the treatment group (P < 0.05).

CONCLUSIONCYD could significantly improve clinical symptoms of ACH patients, decrease serum levels of TBIL and DBIL, reduce serum levels of ALP, GGT, and TBA, obviously improve cholestasis, and promote the recovery.

Alkaline Phosphatase ; Bilirubin ; Cholestasis ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Hepatitis ; drug therapy ; Humans ; gamma-Glutamyltransferase

Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.

Objective To observe the effect of modified Qinghao Biejia Decoction ( QBD) on Th17 cells and renal pathology of MRL/lpr mice with spontaneous systemic lupus erythematosus. Methods Thirty-two female MRL/lpr mice aged 8 to 10 weeks were divided into 4 groups: model group, Chinese medicine group, prednisone group, and combination group, 8 mice in each group. Eight female C57BL/6 mice aged 8 to 10 weeks served as normal control. Mice in Chinese medicine group were given concentrated solution of modified QBD (19.25 g·kg-1·d-1), mice in the prednisone group were given water solution of prednisone acetate (8.75 mg·kg-1·d-1) , mice in the combination group were given the above two kinds of medicine, and mice in the model group and normal control group were given physiological saline. After medication for 7 weeks, spleens and kidneys in all of the groups were taken out for the experiment. Th17 cells in splenic mononuclear cell suspension were detected by flow cytometry, the pathological changes of renal tissue were observed under light microscope, and activity index (AI) of renal tissue in lupus nephritis mice was scored. Results The proportion of Th17 cells in the model group was significantly higher than that of normal control group ( P<0.05) . The proportion of Th17 cells in Chinese medicine group and combination group was lower than that of the model group ( P<0.05) , and prednisone group had higher proportion of Th17 cells than Chinese medicine group ( P<0.05) . Compared with the model group, pathological changes of renal tissue were relieved, and AI scores were decreased in Chinese medicine group, the prednisone group and the combination group ( P<0.05) . Except for the normal control group , AI scores in all groups were positively correlated with the proportion of Th17 cells ( r=0.77, P<0.01) . Conclusion Modified QBD can inhibit the expression of Th17 cells and improve the pathological changes of MRL/lpr mice with lupus nephritis.

Sudden sensorineural hearing loss (SSHL),which is a common and frequently encountered disease,is considered to be a medical emergency in otolaryngology.The prevalence of SSHL is increasing in China.The pathogenesis of SSHL is not clear yet.Microcirculatory disorder of inner ear is considered as one of the most important causes of SSHL.In recent years,several reports have found the levels of serum lipids were changed in patients affected by SSHL.The relationship between SSHL and serum lipids was reviewed to provide new ideas for the diagnosis and treatment of SSHL.

To study the protective effect of Arctigenin in goto-kakizaki (GK) rats combined with hypertension macroangiopathy. Six-week-old GK rats were divided randomly according to blood glucose level into four groups: the model group and low, middle and high dose arctigenin groups (12.5, 25, 50 mg x kg(-1)), with Wistar rats as the normal group. All of GK rats were given high-glucose and high-fat diet. After 16 weeks, GK rats were orally administrated with 10 mg x kg(-1) x d(-1) N-Ω-nitro-L-arginine methyl ester for eight weeks. During the modeling, all of arctigenin groups were orally administrated with different dose of arctigenin twice a day; The model group and the normal group were given solvents. At the beginning, mid-term and end of the experiment, blood glucose was measured. At the end of the experiment, efforts were made to detect blood pressure, collect abdominal aortic blood after anesthesia, fix thoracic aorta after bloodletting to make paraffin sections, observe morphological characteristics and detect the expression of VEGF by immunohistochemistry. According to the results, the blood glucose rose in all GK rats, with no significant difference between the drug group and the model group. At the end of the experiment, the blood pressure significantly increased in GK rats, indicating that Arctigenin could notably reduce the blood pressure in GK rats in a dose-dependent manner. The blood routine test showed increases in both the total white blood cell count and differential blood count, MPV and PDW, abnormal blood platelet parameters and decrease in PLT in GK rats, suggesting that Arctigenin could remarkably reduce the total white blood cell count and differential blood count, MPV and PDW. The thoracic aortic morphological observation revealed obvious endangium lesions in GK rats, demonstrating that Arctigenin could ameliorate the lesion extent. VEGF immumohistochemical staining showed a higher VEGF expression in the model group but lower expression in Arctigenin groups. In conclusion, Arctigenin had a protective effect on aorta in GK rats. Its mechanism may be related to blood pressure lowering, anti-inflammation, improvement in blood platelet function and reduction of VEGF expression.
Animals ; Blood Glucose ; metabolism ; Blood Pressure ; drug effects ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; physiopathology ; Diabetic Angiopathies ; etiology ; metabolism ; physiopathology ; prevention & control ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Furans ; administration & dosage ; Humans ; Hypertension ; etiology ; metabolism ; physiopathology ; prevention & control ; Lignans ; administration & dosage ; Male ; Rats ; Rats, Wistar