OBJECTIVETo observe the effect of Tetramethyl pyrazine (TMP) on the cytokines and inflammatory mediators in the serum and the synovial fluid of collagen-induced arthritis (CIA)rats, and further to investigate its possible mechanisms for treating rheumatoid arthritis (RA).

METHODSType II CIA rat model was established. Rats in the TMP group were administered with TMP at 50 mg/kg and 100 mg/kg, once daily. Dexamethasone at 2.0 mg/kg was intramuscularly injected to those in the Dexamethasone treated group, once daily. Normal saline at 2 mL/kg was given to those in the normal control group and the model group, once daily. All medication was started from the 7th day, lasting to the 35th day. CIA rats' foot swelling degree was observed. Contents of serum IL-1, IL-6, IL-2, NO and PGE2in the synovial fluid were detected by radioimmunoassay and nitrate reduction method.

RESULTSCompared with the normal group, the foot swelling obviously increased, contents of NO and PGE2 in the synovial fluid were obviously elevated in the model group (P < 0.01). Compared with the model group, the foot swelling could be obviously inhibited by 100 mg/kg TMP and Dexamethasone; serum levels of IL-1 and IL-6 obviously decreased, serum IL-2 level obviously increased, contents of NO and PGE, decreased (P < 0.01). TMP 50 mg/kg could obviously inhibit the foot swelling of CIA rats (P < 0.01). There was no statistical difference in other indices (P > 0.05).

CONCLUSIONSTMP at 100 mg/kg showed obvious inhibition on CIA rats. Its inhibitory effect might be correlated to inhibiting activities of endogenous cytokines and the generation of inflammatory mediators in inflammation local regions, improving contents of anti-inflammation cytokines, and inducing the balance of the inflammatory cytokine network.

Animals ; Arthritis, Experimental ; blood ; metabolism ; Dinoprostone ; metabolism ; Female ; Interleukin-1beta ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Nitric Oxide ; metabolism ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Synovial Fluid ; metabolism

The scientific after-class activities in pediatrics can increase the synthetic ability of the medical students,including the manners of occupation,professional level,the skill of communication,clinical practice,serving community,the management of information and the critical thinking.

Objective: To observe the influence of electroacupuncture with different frequencies on rats swimming endurance and sift out the optimal frequency parameter.Methods: Wistar rats were randomly allocated to control group, non-acupoint group and electroacupuncture with low frequency group (EA with 2 Hz group), electroacupuncture with medium frequency group (EA with 15 Hz group), and electroacupuncture with high frequency group (EA with 100 Hz group), acupoints Zusanli (ST 36) and Taichong (LR 3) were punctured and rats load swimming time was used as an index, and the change of swimming endurance time before and after acupuncture was observed. Results: After electro-acupuncture, swimming time was increased in 15 Hz and 100 Hz groups respectively, with the significant differences between before and after acupuncture (P＜0.05). There was also a significant difference in the percentage of increase in swimming time between 15 Hz group and other groups (P＜0.05).Conclusion: It is suggested that electroacupuncture at acupoints Zusanli (ST 36) and Taichong (LR 3) with a frequency of 15 Hz could enhance rats's swimming endurance, and 15 Hz might be an optimal frequency parameter during the electrical stimulation.

Objective To evaluate the possible molecular pathogenesis of intractable temporal lobe epilepsy. The potassium ion channel gene KCNJ4 encodes one of the subfamilies of Kir channels, Kir2.3 subunit, which may play an important role in modulating neuronal excitation. Interference in the function or expression of this gene would cause disturbance of ionic concentrations, thus leading to seizure activity. Methods Reverse transcription polymerase chain reaction (RT-PCR) and Western-blot analysis were used to measure the expression alterations of KCNJ4 mRNA as well as its protein product Kir2.3 channel in temporal cortex samples from patients who had undergone temporal lobectomy for intractable epilepsy (n=12). Tissue from 10 subjects who did not have epilepsy served as controls. Results The expression of KCNJ4 mRNA (0.438?0.178) and its protein Kir2.3 (M 50=0.063) were significantly decreased in epileptic brain compared with the controls (P

Objective To know the mental status of spouses of infertility women and the influence of different conditions on spouse of infertility women, so as to pertinently do the health education well. Methods A questionnaire survey was conducted among spouses of 107 infertility women using the SAS (self-rating anxiety scale). Results The score of SAS of spouses of infertility women was (37.68 ± 7.70). The difference had statistical meaning compared to the normal model in China. There were statistical meanings in anxiety status of spouses of infertility women with different ages, educational degree and the ability of economic acceptance (P < 0.05). Conclusions The individual mental nursing should be given to the infertility women and their spouses according to the different situations of individuals, thereby reducing their anxiety emotion and making them actively cooperate with the therapy and nursing.

OBJECTIVEMutations of the iduronate-2-sulfatase (IDS) gene is the ultimate cause of Hunter syndrome. Clarification of the nature of mutations will create a necessary premise for prenatal gene diagnosis. A mucopolysaccharidosis (MPS) type II patient and his parents from an ethnic minority in Yunnan province were studied to identify their possible mutation in IDS gene to establish the basis for prenatal gene diagnosis.

METHODSThe patient was a boy, 6 years and 10 months old. Urine glycosaminoglycans (GAGs) assay was used for preliminary diagnosis of the patient and his parents with the disease. The three related persons' DNA was extracted and the concentration and purity of the DNA were measured after the urine test results confirmed the diagnosis. Polymerase chain reaction-denaturing high performance liquid chromatography (PCR-DHPLC) analysis was performed to detect the position of the mutation around the hot spots of mutation in exon 9, 3, 8 of the IDS gene. DNA bidirectional direct sequencing was applied to analyze the mutation detected by PCR-DHPLC.

RESULTSThe results of GAGs test showed that in the child with MPS, dermatan sulfate (DS) was positive (+++), heparan sulfate (HS) (+++), chondroitin sulfate (CS) and keratan sulfate (KS) were negative (-); while in his parents none of DS, HS, CS and KS was positive. Abnormal peaks in exon 9 of IDS gene shown by PCR-DHPLC were found in the patient. His mother had heterozygotic peaks. A new frame-mutation (1343-TT) in exon 9 of IDS gene of this patient was confirmed by DNA sequencing. The position where mutation occurred was inside codon 407 (TTT), that means two "T" deleted at position 1343 base pair (1343-TT) in cDNA of the IDS gene, caused a new frame-mutation. It caused elongation of the amino acid chain to a terminal codon TGA at position 429. Thus the peptide chain was shortened from 550 to 428 amino acids. The patient is a hemizygote of the mutation and his mother is a heterozygote.

CONCLUSIONA new frame-mutation (1343-TT) on the IDS gene was identified in this study. The patient is a hemizygote and his mother is a heterozygote. The mutation (1343-TT) resulted in loss of 122 amino acids, which probably caused seriously decreased enzyme activity of IDS, and the authors speculate that this mutation may be the pathological basis of the disease. So, if the mother becomes pregnant again, a prenatal gene diagnostic test for the same mutation should be performed. Furthermore, PCR-DHPLC followed by DNA sequencing are effective methods for diagnosis, including prenatal diagnosis of MPS II.

Amino Acid Sequence ; Asian Continental Ancestry Group ; Base Sequence ; Child ; China ; Chromatography, High Pressure Liquid ; Genotype ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis II ; diagnosis ; genetics ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction

Objective To observe the influence of coal burning fluorosis on learning and memory ability in rats and reveal its possible mechanisms. Methods Healthy 48 SD rats were divided into control, low-fluoride and high-fluoride group. All rats in fluoride exposed groups were fed with the eom polluted by drying processes with burning coal containing high level of fluoride obtained from the endemic fluorosis area to produce the animal model of fluorosis. The experiment period were 3,6 mouths, respectively. The ability of leaning and memory was measured by Morris test and cholinesterase activity detected by photometric method at 3 or 6 month after experiment, respectively. Results Fluoride contents signifieantlly influenced the escape latency, the numbers of crossing the platforms and the time of staying the platforms(the value of F was 29.29,6.47,6.50, respectively, P<0.01).In addition, the numbers of crossing the platforms and the time of staying the platforms were influenced by the exposed time(the value of F was 16.11,45.59, P<0.01). Furthermore, the fluoride contents and the exposed time had an interaction between the numbers of crossing the platforms and the time of staying the platforms (the value of F was 4.67,5.68, P<0.05 or<0.01). Three months after the experiment, the mean values of escape latency [(14.71± 4.85)s] of rats in highly fluoride exposed group were significantly prolonged as compared with controls [(9.28±4.22)s]; 6 month after the experiment, the mean values of escape latency[(12.42±8.03)s, (17.48± 8.05)s] of rats in both groups exposed to fluoride were significantly prolonged as compared to controls [(7.04± 3.29)s, P<0.05]. The decreased numbers of crossing the platforms[(1.62±0.87)number] and the declined time of staying the platforms[(16.70±5.02)s] were found in the rats exposed to high fluoride as compared to controls [(3.53±1.67 )number, (23.33±5.35)s, P<0.05]. The fluoride contents obviously influenced the activities of acetylcholinesterase and butylcolinesterase (the value of F was 12.83,13.27, P<0.01). On the other hand, the times of breeding also influnced the activities of butylcolinesterase (the value of F was 16.26, P<0.01). In 3 months of the experiment, the activities of butylcolinesterase [(0.55±0.12)kU/g] in low fluoride exposed group were significantly decreased in comparison with controls[(0.73±0.10)kU/g, P<0.05]. The activities of acetylcholinesterase[(0.62±0.42)kU/g] and butylcolioesterase[(0.58±0.10)kU/g] in high fluoride group were significantly decreased as compared to eontrois[(1.41±0.52), (0.73±0.10)kU/g, P<0.05]. The correlation analysis showed that there was a negative correlation between the cholinesterase and the escape latency(r=-0.68, P< 0.01), and a positive correlation between the cholinesterase and the time of staying the platforms(r=0.57, P< 0.01). Conclusions The ability of learning and memory in rats with coal buring fluorosis was decreased, which might be connected to the decreased activity of cholinesterase in a dose-effect correlation.

BACKGROUNDBased on in vivo research on the effect of the coating of the extracellular matrix composition of pedicle screws on the conduction and induction of bone formation in young sheep, the aim of this study was to investigate the application of coated pedicle screws in sheep with scoliosis whose spines are under constant development.

METHODSFour groups of pedicle screws were randomly implanted into bilateral L2-L5 pedicles of 2.5- to 3-month-old sheep. A static experiment was performed on one side and a loading test was performed on the other side by implanting connecting rods at the L2-L3 and L4-L5 segments. The changes in the force on the coated screws and the combination of the surface of the coated screws with the surrounding bone in the growth process of young sheep's spines with aging were observed. After 3 months, the lumbar vertebrae with the screws were removed and examined by micro-CT, histological, and biomechanical analyses.

RESULTSUnder nonloading conditions, there is bone formation around the surfaces of coated screws. The bone forming on the surface of collagen/chondroitin sulfate/hydroxyapatite coating of pedicle screws is the most, the one of the collagen/chondroitin sulfate coating and hydroxyapatite coating is followed, and no significant difference between the two groups. In terms of the trabecular bone morphology parameters of the region of interest around the surface of the pedicle screws, such as bone mineral content, bone mineral density, tissue mineral content, tissue bone mineral density, bone volume fraction, and connection density, those associated with collagen/chondroitin sulfate/hydroxyapatite coatings are largest and those unassociated with coatings are smallest. Under nonloading conditions, the pullout strength of the collagen/chondroitin sulfate/hydroxyapatite-coated screws was largest, and that of the uncoated screws was minimal (P < 0.01). Under loading conditions, the maximum pullout strength of each group of pedicle screws was less than that of the pedicle screws in the nonloading state (P < 0.01) with no significant difference between the groups (P > 0.05).

CONCLUSIONSUnder nonloading conditions, the coatings of both organic and inorganic components of the extracellular matrix of titanium pedicle screws can conduct or induce bone formation around the surface of the screws. The ability of collagen/chondroitin sulfate/hydroxyapatite coatings to induce bone formation is stronger; under loading conditions, a large amount of connective tissue formed around the surfaces of the screws in each group. No significant differences were found between the groups.

Animals ; Biomechanical Phenomena ; Bone Screws ; Chondroitin Sulfates ; chemistry ; Collagen ; chemistry ; Durapatite ; chemistry ; Rats ; Sheep ; X-Ray Microtomography

OBJECTIVETo investigate flap survival after transfection using gene encoding vascular endothelial growth factor (VEGF).

METHODSIn 30 Sprague-Dawley rats, the anterior abdominal skin flap supplied by the epigastric vessels was created. The animals were divided into three groups, with ten of each. The first group was treated with a mixture of liposomes and the cDNA encoding the 165-amino acid isoform of VEGF; the second group was treated with control blank plasmid DNA and liposome transfection medium; the third group was treated with physiological saline. Four days after injection, the epigastric artery and vein were ligated and the blood flow in the flap was evaluated by intraperitoneal injection of fluorescence solution. Seven days later, the survival area of the flap was measured by planimetry. After the animals were killed, specimens were harvested from the anterior abdomen skin flap for immunohistological evidence of VEGF expression and for hematoxylin and eosin staining of microvascular growth.

RESULTS30 minutes after pedicle ligation the average fluorescence staining planimetry of the three groups (PCD-VEGF165, PCD and physiological saline) was 60.64%, 30.15% and 29.89% respectively. Tissue survival planimetry of the three groups was 92.3%, 30.5%, 31.8%. There was significant difference between the first group and the latter two (P < 0.05). Immunohistochemical staining documented increased deposition of VEGF cDNA in the first group compared to the control groups (P < 0.05). Normal staining documented that the average vessel number of the three groups was 101.72, 91.35 and 89.85 (P < 0.05), the average vessel lumen diameter was 26 microns, 31.09 microns and 32.51 microns(P < 0.05).

CONCLUSIONThrough administration, PCD-VEGF165 can transfect the anterior abdominal skin flap and enhance its survival. There was express of VEGF protein in the treated flap.

Animals ; Biomarkers ; metabolism ; DNA, Complementary ; administration & dosage ; Epigastric Arteries ; Genetic Therapy ; Graft Survival ; physiology ; Liposomes ; Plasmids ; Rats ; Rats, Sprague-Dawley ; Surgical Flaps ; blood supply ; physiology ; Time Factors ; Transfection ; methods ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the effects of high temperature stress on the root vitality and leaf biochemical indexes in populations of Pinellia ternate.

METHODThe leaf activity of SOD, contents of MDA and free praline and root vitality were determined after a treatment of high temperature stress (35 degrees C/25 degrees C) was given to different populations of P. ternate.

RESULT AND CONCLUSIONBy stress time increasing, the leaf SOD and free praline rose firstly and then dropped, the content of MDA increased while the root ability decreased in all populations. And the response of populations of P. ternate to high temperature was significantly different.

China ; Ecosystem ; Heat-Shock Response ; physiology ; Hot Temperature ; Malondialdehyde ; metabolism ; Pinellia ; metabolism ; physiology ; Plant Leaves ; metabolism ; physiology ; Plant Roots ; physiology ; Plants, Medicinal ; metabolism ; physiology ; Proline ; metabolism ; Superoxide Dismutase ; metabolism