BACKGROUND: Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Researches suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.OBJECTIVE: To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 (IL-1),IL-6 and tumor necrosis factor alpha (TNF-α) in aqueous humor of rabbits after extracapsular cataract extraction.DESIGN: Randomized controlled animal study.SETTING: Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.MATERIALS: The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes,general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification: SYXK (yue) 2005-006]. Eyes of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawei Biotechnology Company Limited.METHODS: Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mi/kg urethan, and then, 5# needle was punctured from corneal limbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque: horizontally intercepting bladder or waterly separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1st, 3rd, 7th and 14th days after operation, inflammatory reaction of anterior chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meanwhile, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.MAIN OUTCOME MEASURES: Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rabbits in three groups after operation.RESULTS: ① On the 1st, 3rd, 7th and 14th days after operation, numbers of leucocytes were (2.4±0.7)×106/L, (2,2±0.5)×106/L, (2.8±0.8)×106/L and (2.0±0.5)×106/L in aqueous humor; (19.7±7.3)×106/L, (28.1±9.6)×106/L, (14.2±5.6)×106/L and(8.4±3.8)×106/L in traumatic control group; (65.3±14.5)×106/L, (79.8±12.7)×106/L, (21.7±8.2)×106/L and (12.4±4.1)×106/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group (F =22.5, 27.9, 11.6, 8.4;P＜0.05). ② Within 1-14 days after operation, contents of IL-1, IL-6and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group (P＜0.05), and there was a significant difference between traumatic control group and operative group (P＜0.05);however, there was no significant difference among three groups on the operative day (P＞0.05). ③ Contents of cytokines reached peak on the 7th day after operation, decreased gradually, and reached the lowest value on the 14th day.Contents in traumatic control group and operative group were higher than those in normal control group (P＜0.05).CONCLUSION: The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism ; Saponins ; administration & dosage

Objective To analyze the effect of probiotics combined with early enteral nutrition on infection and gastrointestinal dysfunction in patients undergoing mechanical ventilation in intensive care unit (ICU).Methods Prospective cohort study was adopted to select patients who admitted to ICU in a hospital from February 2016 to October 2017, they were randomly divided into three groups:A, B, and C.Group A received early enteral nutrition combined with probiotics, group B received early enteral nutrition, and group C received early parenteral nutrition.Infection condition, level of infection indicators (on the 3 rd, 7 th and 14 th day after treatment), occurrence of gastrointestinal dysfunction, acute physiology and chronic health evaluation II (APACHE II) on the 14 th day after treatment were compared among three groups.Results Incidences of infection in group A, B, and C were 6.00％, 20.00％, and 22.00％ respectively, difference among three groups was significant (χ2=8.57, P=0.01).C-reactive protein (CRP) and white blood cell count (WBC) in group A on the 7 th and 14 th day were both lower than those in group B and C;procalcitonin (PCT) in group A and B on the 3 rd day were both lower than that in group C;PCT in group A on the 7 th and 14 th day were both lower than those in group B and C;difference were all statistically significant (all P＜ 0.05).Incidence of abdominal distension (8.00％), diarrhea (4.00％) and gastric retention (4.00％) in group A were the lowest among three groups.APACHE II score in group A on the 14 th day after treatment was lowest.Conclusion Early enteral nutrition combined with probiotics for treatment of ICU patients with mechanical ventilation can effectively reduce the incidence of infection and gastrointestinal dysfunction, promote rehabilitation, which is worth promoting the application.

Ten compounds were isolated from the barks of Jasminum giraldii by means of various of chromatographic techniques such as silica gel, Sephadex LH-20 and Rp-HPLC. Their structures were identified by spectroscopic data analysis as (+)-medioresinol (1), (+) -syringaresinol (2), syringaresinol-4'-O-beta-D-glucopyranoside (3), oleanic acid (4), 3-methoxy-4-hydroxy-trans-cinnamaldehyde (5), trans-sinapaldehyde (6), syringaldehyde (7), 1-(4-methoxy -phenyl) -ethanol (8), trans-cinnamic acid (9), and 4-(1-methoxyethyl) -phenol (10). Among them, compounds 1-3, 5-8 and 10 were isolated from the J. genus for the first time and compounds 4 and 9 were obtained from J. giraldii for the first time. In the DPPH free radical scavenging assay, compound 1 exhibited significant activity (IC50 55.1 micromol x L(-1)), compared with vitamin C(IC50 59.9 micromol x L(-1)); and compound 2 showed moderate activity (IC50 79.0 micromol x L(-1)), compared with 2, 6-di-tert-butyl4-methylphenol (IC50 236 micromol x L(-1)).
Antioxidants ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Jasminum ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the effect of carbon disulfide (CS(2)) on oxidation-antioxidation function of rat nerve tissues.

METHODSThirty male Wistar rats were randomly divided into the control group, the low-dosage exposure group and the high-dosage group, 10 rats each. The rats of the two exposure groups were administered with CS(2) by gavage at a dosage of 300 or 500 mgxkg(-1)xd(-1), 5 times every week for continuous 12 weeks. The alterations in glutathione (GSH), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), hydrogen peroxidase (CAT) and total anti-oxidation (T-AOC) in cerebrum, spinal cord, and sciatic nerve of CS(2)-treated animals were assayed.

RESULTSThe results showed that the contents of MDA and ROS in nerve tissues of CS(2)-treated groups increased significantly except ROS in spinal cord and sciatic nerve of low dose group. The content of MDA was increased by 20.7% and 33.6% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 18.5% and 23.3% respectively in the spinal cord, and by 20.7% and 53.0% respectively in the sciatic nerve, The content of MOS was increased by 20.1% and 34.9% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, and by 14.1% and 15.4% respectively in the spinal cord and the sciatic nerve of the rats of the high-dosage group (P < 0.05 or P < 0.01). Furthermore, the activities of SOD, GSH-Px, CAT and T-AOC decreased significantly except GSH-Px and SOD in cerebrum of low dose group. The content of GSH was decreased by 17.2% and 26.5% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 26.4% and 31.2% respectively in the spinal cord, and by 15.1% and 20.0% respectively in the sciatic nerve. The content of T-AOC was decreased by 11.1 and 26.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 15.1% and 38.4% respectively in the spinal cord, and by 35.6% and 42.3% respectively in the sciatic nerve. The activity of SOD was decreased by 12.1% and 25.4% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 16.4% and 30.3% respectively in the sciatic nerve. The activity of GSH-Px was decreased by 17.3% and 32.5% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 17.1% and 21.5% respectively in the sciatic nerve. The activity of GSH-Px and SOD was decreased by 12.6% and 30.1% respectively in the cerebrum of the rats of the high-dosage group. The activity of CAT was decreased by 17.5% and 39.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 25.2% and 31.3% respectively in the spinal cord, and by 17.1% and 36.9% respectively in the sciatic nerve (P < 0.05 or P < 0.01).

CONCLUSIONSubchronic exposure to CS(2) can induce significant changes of oxidation-antioxidation function in rat nerve tissues, which might be related to CS(2)-induced neurotoxicity.

Animals ; Antioxidants ; metabolism ; Carbon Disulfide ; Lipid Peroxidation ; drug effects ; Nerve Tissue ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo observe beta-catenin expression of Wnt signaling pathway in rats with knee osteoarthritis, and influence of Bushen Huoxue decoction on beta-catenin and MMP-7 expression of synoviocytes in rats with knee osteoarthritis (OA).

METHODSRats model with knee osteoarthritis were established by Hulth method. Primary synoviocytes and OA synoviocytes were cultured with collagenase digestion method. The cultured synoviocytes were divided into normal group, OA model group and Bushen Huoxue decoction group. Western blotting method was used to detect beta-catenin, MMP-7 protein expression of synoviocytes after acting by Bushen Huoxue decoction for 48 h; ELISA method was used to detect MMP-7 expression of synovial supernatant.

RESULTSOA synoviocytes were cultured successfully. Western blotting showed that beta-catenin, MMP-7 expression in OA synoviocytes was significantly higher than normal group, Bushen Huoxue decoction could significantly reduce beta-catenin, MMP-7 expression; ELISA results showed that MMP-7 expression of OA synovial supernatant was significantly higher than normal synoviocytes supernatant, Bushen Huoxue decoction significantly regulated the level MMP-7 down.

CONCLUSION(1) High expression of beta-catenin in OA synoviocytes indicates that Wnt classical signal pathway is activated in rat with knee osteoarthritis; (2) High expression of MMP-7 expression in OA synoviocytes confirms the MMP-7 is downstream genes of Wnt/beta-catenin signal pathway; (3) Activation of Wnt signal pathway and increase of MMP-7 may cause degradation of articular cartilage, and promote the formation of osteoarthritis; (4) Bushen Huoxue decoction can reduce expression of MMP-7, and promote cartilage repair, which may be one of mechanisms of osteoarthritis.

Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Male ; Matrix Metalloproteinase 7 ; analysis ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Synovial Fluid ; chemistry ; cytology ; beta Catenin ; analysis

OBJECTIVETo explore the influence of knock-down of miR-21 expression on the radiosensitivity of radioresistant glioma SHG-44 cells and its mechanism.

METHODSRadioresistant cell line SHG-44(R) cells were established from radiosensitive parental SHG-44 glioma cells. Then the expression levels of miR-21 in SHG-44(R) and SHG-44 were determined by quantitative real-time PCR. The effect of miR-21 on the radiosensitivity was assessed in SHG-44(R) with miR-21 inhibitor to decrease the miR-21 expression.

RESULTSmiR-21 was up-regulated 1.49-fold in SHG-44(R) cells relative to the SHG-44 cells. But after transfection with As-miR-21, the miR-21 expression in SHG-44(R) cells was knocked down significantly. As-miR-21 combined with radiation could synergistically enhanced mitotic death and apoptosis of SHG-44(R) cells, with SER of D(0) and D(q) being equal to 1.48 and 1.54, respectively. Expression levels of caspase-3 in the radiation group, AS-miR-21 transfection group and combination group were 2.24 ± 0.14, 2.05 ± 0.19 and 5.72 ± 0.45, respectively, and its expression in the combination group was higher than that in the other two groups.

CONCLUSIONSmiR-21 may be involved in the formation of radioresistance of glioma cells and as a potential target for enhancing the response of radioresistant-glioma cells to radiotheapy.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Glioma ; genetics ; metabolism ; pathology ; Humans ; MicroRNAs ; genetics ; metabolism ; Oligonucleotides, Antisense ; genetics ; metabolism ; Radiation Tolerance ; Transfection

Osteoarthritis is a degenerative joint disease,with the characters of degradation of articular cartilage, the formation of the joint marginal osteophyte and synovium lesions. Previous studies have focused on the treatment of articular cartilage lesions. In recent years, new research in shows synovial inflammation plays an important role in OA. Synovium lesions and synovial inflammation-related factors induced the degradation and destruction of articular cartilage, and promoted the development of osteoarthritis. The role of synovial lesions in osteoarthritis is increasingly prominent, and the treatment for synovial lesions will become a new target. So this paper reviews the various manifestations of synovial in osteoarthritis.
Humans ; Osteoarthritis ; pathology ; Synovial Membrane ; pathology

OBJECTIVETo evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms.

METHODSSixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-β1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing.

RESULTSIntraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-β1 and collagen I expressions in the ischemic tissues.

CONCLUSIONSIntraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-β signaling pathway.

Animals ; Cecum ; injuries ; pathology ; Collagen Type I ; metabolism ; Fibrinolysis ; Injections, Intraperitoneal ; Peritoneum ; injuries ; pathology ; Phenanthrenes ; pharmacology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Postoperative Complications ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Adhesions ; prevention & control ; Tissue Plasminogen Activator ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing