OBJECTIVETo observe the changes of bone histomorphometry parameters in condyle of rabbits during mandibular forward positioning.

METHODSForty rabbits with eight weeks of age were simple randomly divided into the experimental group (n = 24) and control group (n = 16).Mandibles of rabbits in the experimental groups were induced to forward position by a functional appliance.The experimental group and control group were sacrificed after 2 and 4, 8, 12 weeks.The specimens from right tempromandibular joints were processed for undemineralised sections.These sections were used for fluorescent microscopy observation and the measurement of bone histomorphometry.

RESULTSAt 8 week bone-volume fraction [(75.83 ± 1.10)%], trabecular thickness [(103.28 ± 2.89) µm], trabecular number [(2.86 ± 0.06) mm(-1)], mineral apposition rate [(2.32 ± 0.02) µm/d] and index of osteoblast [(30.20 ± 0.47) N/mm(2)] in subchondral region of the cartilage in the experimental group were significantly increased compared with the age-matched controls [respectively (64.00 ± 1.54)%, (87.00 ± 1.13) µm, (1.84 ± 0.08) mm(-1), (1.69 ± 0.02) µm/d and (21.07 ± 0.59) N/mm(2)] (P < 0.05). However, trabecular separation [(170.00 ± 2.25) µm] was lower than those in the controls [(241.50 ± 1.57) µm] with significant difference(P < 0.05). There was no significant difference in the bone histomorphometry parameters of the central region between the experimental group and the age-matched control group (P > 0.05).

CONCLUSIONSThe pattern of bone histomorphometry parameters expression in subchondral region has a high correlation with the adaptive remodeling of the condyle after functional appliance.

Animals ; Cartilage ; anatomy & histology ; growth & development ; Female ; Male ; Mandibular Advancement ; Mandibular Condyle ; anatomy & histology ; growth & development ; Orthodontic Appliances, Functional ; Rabbits ; Random Allocation

OBJECTIVETo study c-fos and substance P expression in the central nervous system following mechanical and chemical nociceptive stimulation to the masseters in rats with occlusal interference.

METHODSOcclusal interference was made by bonding a 2 mm long dentin screw in the pulp cavity of the first maxillary molar in the left side. Seven days after occlusal interference, the rats in occlusal interference and mechanical stimulus group and mechanical stimulus control group were light anesthetized and nociceptive mechanical stimulus were applied to the ipsilateral masseter. Pain response was recorded and all the animals were killed 2 hours later. The rats in the other two groups were deep anesthetized and 100 microL 5% formalin was injected into the ipsilateral masseter, killed 2 hours later. The brainstem and cervical spinal cord were processed c-fos and substance P immunoreactivity and data were quantitatively analyzed.

RESULTSBoth mechanical and chemical stimulus to the ipsilateral masseter induced increasing neuronal c-fos expression in the trigeminal nucleus and in the cervical spinal dorsal horn in occlusal interference and mechanical stimulus group and occlusal interference and chemical stimulus group (P < 0.05). Following mechanical stimulation to the ipsilateral masseter, substance P expression in the trigeminal nucleus transition zone was increased in occlusal interference and mechanical stimulus group (P < 0.05).

CONCLUSIONThe central neuronal sensitization in the brainstem may play an important role in the masticatory muscle pain induced by occlusal interference.

Animals ; Masseter Muscle ; Masticatory Muscles ; Pain ; Proto-Oncogene Proteins c-fos ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of apoptosis in the remodeling of temporomandibular joint (TMJ) under pressure.

METHODSSynovial fibroblasts obtained from rat temporomandibular joint were subjected to different hydrostatic pressure for 12 h. Changes of ultrastructure were observed by transmission electron microscope.

RESULTSAt 30 kPa, the ultrastructure of synovial fibroblasts showed no obvious changes. At 60 kPa, the chromatin was condensated, the baryon took on crescent and the mitochondria seemed varicose. At 90 kPa, the apoptosis-like body was wrapped by membrane and embedded in the high density chromatin.

CONCLUSIONSApoptosis-like change took place in ultrastructure of synovial fibroblasts under hydrostatic pressure, and the degree of the change was related to the hydrostatic pressure exerted.

Animals ; Apoptosis ; Fibroblasts ; ultrastructure ; Hydrostatic Pressure ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; cytology ; Temporomandibular Joint ; cytology

ObjectiveTo investigate the regulatory effects of traditional Chinese medicine (TCM) Shufengxuanfeijiedu formula on Janus kinase signal transducer and activators of transcription (JAK-STAT) of lung tissues in mice with influenza viral pneumonia.Methods According to random number table, 60 mice were randomly divided into six groups with 10 mice in each group: normal group (N), model group (M), Tamiflu control group (C) and low (SL), medium (SM), high dose (SH) Shufengxuanfeijiedu formula groups. The mouse model of influenza virus pneumonia was reproduced by dropping of 0.05 mL 4LD50 inflluenza virus FM1 strain which can be adapted to lung tissue into the nose; while the N received nose instillation of 0.05 mL normal saline. After successful modeling for 2 hours, distilled water was given orally (by lavage) to N and M; Duffy (oseltamivir) 2.5 g·mL-1·d-1 was administrated to C; the TCM SL, SM, SH were intragastrically administered with different doses of shufengxuanfeijiedu decoction into the corresponding groups respectively (the ingredients of prescription: chrysanthemum, mulberry leaf, almond, platycodon root, forsythia, bupleurum etc. forming granules), according to the suitable dose of granules used for human body surface, the dose used for mouse surface area was calculated, the high dose means the dose used in the medium dose group doubled, the low dose means 1/2 dose used in medium group, once a day, once 0.2 mL for consecutive 4 days. Afterwards, the lung tissues were collected, the mouse differential gene expressions related to JAK-STAT pathway were detected by gene chip technology, the standards for screening of differential gene expression were as follows: up-regulated gene was P < 0.05, and the log2ratio > 1; down-regulation gene wasP < 0.05, and log2ratio < -1. The levels in lung tissue kinase (JAK) andγinterferon (IFN-γ) mRNA expressions were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR).Results Compared with those in N, the differential expression gene transcription activator, STAT5 [log2 (N/M) = 2.32], interleukin 4 receptor alpha subunit [IL4RA, log2 (N/M) = 4.77], interleukin 12 receptor [IL12R, log2 (N/M) = 1.58], JAK [log2 (N/M) = 2.41] were all obviously up-regulated, and IFN was significantly down-regulated [log2 (N/M) = -1.45] in M. Compared with those in M, C group IFN [log2 (C/M) = 1.51], various TCM dose groups [log2 (SL/M) = 1.46, log2 (SM/M) = 1.72, log2 (SH/M) = 1.40] differential expression gene IFN was significantly up-regulated, STAT5 [log2 (C/M) = -2.06, log2 (SL/M) = -1.41, log2 (SM/M) = -2.10, log2 (SH/M) = -1.89], IL4RA [log2 (C/M) = -2.52, log2 (SL/M) = -1.85, log2 (SM/M) = -2.74, log2 (SH/M) = -1.39), IL12R [log2 (C/M) = -1.48, log2 (SL/M) = -0.10, log2 (SM/M) = -1.58, log2 (SH/M) = -0.53], JAK [log2 (C/M) = -1.44, log2 (SL/M) = -0.88, log2 (SM/M) = -1.74, log2 (SH/M) = -0.53] were significantly down-regulated. In M, the JAK mRNA expression was obviously elevated (2-ΔΔCt: 3.17±0.94 vs. 1.01±0.13,P < 0.05), while the IFN-γ mRNA expression was decreased (2-ΔΔCt: 0.15±0.48 vs. 1.01±0.12,P < 0.05); compared with M, the JAK mRNA expressions in C, SM and SH groups were all obviously decreased (2-ΔΔCt: 2.02±0.63, 1.19±0.30, 1.59±0.67 vs. 3.17±0.94, allP < 0.05); while the IFN-γmRNA expressions in C, SL, SM and SH groups were elevated (2-ΔΔCt: 0.61±0.12, 0.41±0.13, 0.85±0.14, 0.78±0.20 vs. 0.15±0.48, allP < 0.05).Conclusions Shufengxuanfeijiedu formula can ameliorate the mice immune pathological injury of lung tissues induced by influenza virus by regulating JAK-STAT signal pathway and balancing Th1/2 via up-regulating the expression of IFN-γ.

Objective To evaluate the feasibility of 3-(4-~(18)F-fluorobenzyl)-8,9-dimethoxy-1,2,3,4-tetrahydrochromeno [3,4-c]pyridin-5-one ( is F-FDTP) as a potential dopamine D4 receptor PET imaging agent.Methods ~(18)F-FDTP solution in ethanol-physiological saline was incubated with calf serum to test its in vitro stability through the determination of radiochemical purity.Normal rats were injected intravenously with ~(18)F-FDTP and then sacrificed at 2,5,10,15,30,60 and 120 min after anesthesia.Blood,organs and brain tissue samples were collected.All samples were weighed and measured for radioactivity.The uptake of samples was expressed as percentage activity of injection dose per gram of tissue ( % ID/g).Results The stability of ~(18)F-FDTP was satisfactory and its radiochemical purity was above 95% after incubation 120 min at 37℃ in calf serum.The biodistribution showed that ~(18)F-FDTP could penetrate through the blood-brain barrier and selectively accumulate in striatum,hypothalamus,frontal certex,hippocampus,cerebellum,where the D_4 receptor was reportedly located.The radioactivities in hippocampus,hypothalamus,striatum,frontal cortex,cerebellum,pons were (0.42±0.03),(0.46±0.05),(0.54±0.04),(0.39±0.04),(0.45±0.06),(0.35±0.04) %ID/g,respectively,2 min post injection.And there was difference between the normal biodistribution results and the blocking experimental results:(0.36 ±0.05),( 0.33±0.05 ),(0.55±0.05 ),(0.30±0.07 ),(0.34±0.07 ) and (0.32±0.04) % ID/g in hippocampus,hypothalamus,striatum,frontal cortex,cerebellum and pons,respectively.Conclusions ~(18)F-FDTP can penetrate through the blood-brain barrier and selectively accumulate in striatum,hypothalamus,frontal cortex,hippocampus,cerebellum,where the D_4 receptor was known to concentrate.These preliminary results suggest that ~(18)F-FDTP is a potential dopamine D_4 receptor imaging agent and further studies are needed.

Objective To study the influences of mental disorders on female systemic lupus erythematosus(SLE)and analyze the factors. Methods We used symptom check list -90 (SCL-90) as a basis for judging mental disorders disease activity. Disease activity, social support and depreciation - discrimination were used as possible influencing factors. Social support and discomfort – discrimination were possible influencing factors. Multivariate unconditional logistic regression model was used to analyze the influencing factors of mental disorders. Results The total score of SCL-90 of patients with female SLE was significantly higher than that of norm models ［(136.39±48.66) vs (129.96±38.76)］ (P<0.05), in 289 SLE patients, the number of patients with mental disorders was 128 (44.3%). High monthly income(OR=0.770, 95% CI:0.604-0.981, P=0.034) was a protective factor for mental disorders. High disease activity (OR=1.792, 95% CI:1.023-3.138, P=0.042)and high discomfort–discrimination (OR=1.100, 95% CI:1.035-1.169, P=0.002)were risk factors for mental disorders. Conclusions Female SLE patients have a higher risk of mental disorders than the general population. And eliminating self-depreciation, reducing social discrimination, active employment, increasing monthly income, standardizing treatment and reducing disease activity may effectively alleviate mental disorders in SLE patients.

OBJECTIVETo construct the recombinant vectors that express mouse Dishevelled 2 (Dvl2), and to evaluate its expression level in transfected RAW264.7 cells.

METHODSA pair of specific primers were designed according to the mouse Dvl2 cDNA sequence published in GenBank. Total RNA of RAW264.7 cells was extracted, and open reading frame of Dvl2 was obtained by RT-PCR, which was then cloned into pEZ-M29 plasmid. Electrophoresis after macrorestriction and DNA sequence analysis were used to identify the reconstructed plasmids. After transient transfection via liposome, the transfection of RAW264.7 cells was confirmed under a fluorescence microscope, and the expression level of Dvl2 was evaluated by real-time RT-PCR.

RESULTSThe recombinant plasmid containing mouse Dvl2, namely pEZ-M29/ Dvl2, was successfully constructed, and confirmed by DNA sequence analysis. After 48 h of tranfection, the expression of enhanced green fluorescene protein was observed under a fluorescence microscope, and real-time RT-PCR analysis revealed that the Dvl2 mRNA level was prominantly elevated in the transient transfected RAW264.7 cells.

CONCLUSIONThe recombinant plasmids pEZ-M29/Dvl2 are successfully constructed and can elevate the Dvl2 mRNA level in the transient transfected RAW264.7 cells, which can be used in further studies aiming at revealing the functional significance of Dvl2 in the osteoclastogenesis.

Adaptor Proteins, Signal Transducing ; biosynthesis ; Animals ; Cell Line, Tumor ; Dishevelled Proteins ; Genetic Vectors ; Mice ; Phosphoproteins ; biosynthesis ; Plasmids ; RNA, Messenger ; Transfection

OBJECTIVETo study the adaptive alteration in bilaminar zone of rabbits' temporomandibular joint following disc displacement.

METHODSTwenty-six Japanese white rabbits were used in this study. Among these rabbits,6 were used as controls. The right discs of other 20 rabbits were displaced anteriorly by operation. Four of these rabbits were killedatn 1, 2, 4, 6 and 8 weeks respectively after surgery. The TMJS were studied by HE staining, Alcin bluen staining and in situ detection of type II collagen mRNA expression.

RESULTSThere appeared cartilage metaplasia after one week following disc displacement. Typical chondrocytes could be found in the bilaminar zone. The new chondrocytes expressed type II collagen.

CONCLUSIONSThe bilaminar zone of TMJ will be remodeled following disc displacement and become a disc-like tissue to function as a disc.

Animals ; Collagen Type II ; biosynthesis ; genetics ; Female ; Joint Dislocations ; metabolism ; Male ; RNA, Messenger ; biosynthesis ; Rabbits ; Temporomandibular Joint Disc ; metabolism ; pathology ; Temporomandibular Joint Disorders ; metabolism ; pathology

OBJECTIVETo study the p53/p21 fusion gene as a potential fusion gene for the gene therapy of human oral squamous cell carcinoma.

METHODSp21 cDNA was obtained from normal human embryonic lung cells by RT-PCR, fusing with p53 gene. The recombinant plasmid pcDNA-p53/p21 was constructed by inserting the p53/p21 fusion gene into eukaryotic expression vector pcDNA3.1 and subsequently transfected into human oral squamous cell carcinoma cell line (Tca8113) with lipofectamine. RT-PCR and Western blot were used to demonstrate the expression of p53/p21 fusion gene. Using clonal formation experiment and (3)H-TdR incorporation assay were used to evaluate the clonal formation and proliferation ability of Tca8113 cells.

RESULTSIt was observed that p53/p21 fusion gene could inhibit clonal formation and proliferation of human oral carcinoma. RT-PCR and Western blot demonstrated that it was the expression of exogenous p53/p21 fusion gene that led to the above results.

CONCLUSIONSTransfection of p53/p21 fusion gene to Tca8113 cells could inhibit the tumor cell proliferation and clone formation in vitro, and make itself a potential fusion gene for the gene therapy of human oral squamous cell carcinoma.

Carcinoma, Squamous Cell ; therapy ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Gene Fusion ; genetics ; Genes, p53 ; genetics ; Genetic Therapy ; Humans ; Mouth Neoplasms ; therapy

The characteristics of purinoceptors in the membrane of rat trigeminal ganglion (TG) neurons were studied by using whole- cell patch clamp technique. The results showed that most of neurons examined (78.9%, 142/180) were responsive to ATP in a concentration-dependent manner; the others (21.1%, 38/180) were ATP insensitive. Of the ATP-sensitive cells, the majority (95.1%, 135/142) responded to ATP with an inward current, a few (2.1%, 3/142) with an outward current, and the rest (2.8%, 4/142) with biphasic current. Small sized cells (<30 mum) responded to ATP with a rapid desensitizing inward current and were highly sensitive to vanilloid; the medium sized cells (30~40 mum) responded to ATP with slow desensitizing inward current and were not sensitive to vanilloid; while the majority of large sized cells (>40 mum) did not respond to ATP and vanilloid. The waveform of ATP-activated inward currents was related to the cell diameter. The I-V curves for both small and medium sized cells manifested obvious inward rectification. Furthermore, we studied the kinetic features of ATP-activated currents and the effects of P2 purinoceptor agonists and antagonists on I(ATP). The findings suggest that ATP receptor-ion channels are expressed differently among different types of rat TG neurons.
Adenosine Triphosphate ; metabolism ; Animals ; Animals, Newborn ; Neurons ; metabolism ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X ; physiology ; Trigeminal Ganglion ; cytology ; metabolism ; physiology