Clinical Application of CT Navigation in treatment of Lumbar Spondylolisthesis with Minimally Invasive Surgery - Transforaminal Lumbar Interbody Fusion.

OBJECTIVE: To explore the clinical effect of the application of CT navigation in the treatment of lumbar spondylolisthesis with minimally invasive surgery - transforaminal lumbar interbody fusion (MIS-TLIF). METHODS: A retrospective study was conducted on 30 patients with lumbar spondylolisthesis who were continuously treated in linyi central hospital from May 2018 to March 2019.The patients were divided into two groups,15 patients treated with MIS-TLIF with the aid of CT navigation during the operation were included into an observation group. Another 15 patients were treated with open transforaminal lumbar interbody fusion as the control group. The baseline information, including gender, age and course of disease, perioperative period and imaging conditions, and VAS and ODI scores of patients in the two groups were collected and analyzed. RESULTS: Fifteen patients were included into the observation group, including 9 male and 6 female patients with an average age of 52.60 ± 6.31 and a course of disease of 16.33 ± 6.00 months. The other 15 patients were included into the observation group, including seven male and eight female patients with an average age of 52.87 ± 7.38 and a course of disease of 19.13 ± 9.89 months. The difference in the gender, age and course of disease between the two groups had no statistical significance (P > 0.05). However, the difference in the duration of operation and intraoperative blood loss between the two groups had statistical significance (P< 0.05). There were no statistically significant differences in wound complications, neurological complications, preoperative slippage rate, postoperative slippage rate, slippage reduction rate and screw placement accuracy (P > 0.05). VAS scores of the two groups were statistically significant from six months after surgery (P < 0.01). There was no significant difference in ODI between the two groups at any time point (P >, 0.05). VAS and ODI scores were improved at each time point compared with those before surgery. CONCLUSION: The minimally invasive transforaminal lumbar fusion performed with the aid of CT navigation during the operation shortens the duration of operation and the amount of bleeding, reduces the back pain, is beneficial to the early postoperative functional exercise, and speeds up the postoperative recovery.

Common single nucleotide polymorphisms in cyclooxygenase-2 and risk of severe chronic periodontitis in a Chinese population.

AIM: Several common single nucleotide polymorphisms (SNPs) of the cyclooxygenase-2 (COX-2) gene have been reported to be functional. The association between -1195GA, -765GC and 8473TC of COX-2, and severe chronic periodontitis (CP) in a Chinese population was investigated. MATERIAL AND METHODS: 148 cases of healthy controls (control group) and 146 cases of severe CP were recruited in this study. Genotypes of -1195GA, -765GC and 8473TC were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The distributions of genotypes and haplotypes were compared by chi(2) test and the odds ratios (ORs) were calculated by logistic regression analysis. RESULTS: The prevalence of the -1195A was more prevalent in CP group (60.62%) than control group (51.35%), and the distributions of the -765C and 8473C were higher in control group (6.76% and 21.96%) compared with CP group (3.08% and 15.07%). Only genotype distribution of -1195GA was significant when p-value was corrected for multiple testing (p(c)=0.033). The adjusted ORs for the -1195AA/GA, -765GC and 8473CC/TC were 2.49 (95% CI=1.33-4.69, p=0.005), 0.45 (95% CI=0.20-1.04, p=0.061) and 0.67 (95% CI=0.41-1.11, p=0.118). Subjects with the haplotype AGT had a significantly higher risk of periodontitis than those with the most common haplotype GGT (OR=1.91, 95% CI=1.32-2.76, p(c)<0.001). CONCLUSIONS: It suggests the -1195A variant is associated with an increased risk for severe CP.

[Impacts of periodontitis on visceral organ weight and weight percentage in obese mice].

OBJECTIVE: This study aimed to explore the impacts of periodontitis on the visceral weight and weight percentage of obese animal models. METHODS: A total of 64 C57BL/6J mice were divided into the following diet groups: high-fat diet (HFD) group (n=36), which was fed with high-fat diet to induce obesity, and low-fat diet (LFD) group (n=28), which was fed with low-fat diet as the control. After 16 weeks on diet, each diet group was divided into periodontitis (P) and control (C) groups. The P groups were induced for periodontitis by ligation with Porphyromonas gingivalis-adhered silk for 5 or 10 days, and the C groups were sham-ligated as the control. Visceral organs were resected and weighed. The organ weight percentage was calculated. RESULTS: Compared with the LFD group, the HFD group significantly upregulated the weight and weight percentage of visceral adipose tissue and spleen (P<0.05), upregulated the weight of liver and kidney (P<0.05), and downregulated the weight percentage of liver and kidney (P<0.01). In the HFD group, the weight and weight percentage of spleen were downregulated in the P group (P<0.05), but were upregulated in the 10-day group compared with the 5-day group (P<0.05). CONCLUSIONS: Periodontitis can affect the general morphology of the viscera (especially spleen) in obese animal models. Pathological indications in terms of immunometabolism might be present in the correlation between obesity and periodontitis.

[Beneficial effect of periodontal therapy on insulin resistance and lipid metabolism in obese rats with periodontitis].

OBJECTIVE: To investigate the effect of periodontal therapy in controlling periodontitis and on insulin resistance and lipid metabolism in obese rats with periodontitis. METHODS: Sprague-Dawley rats were randomized into normal group (group C), obese group (group O), periodontitis combined with obesity group (group P) and periodontal treatment group (group T). The obese rats in groups P and T were subjected to ligation of the maxillary second molar with silk thread to induce experimental periodontitis, and the rats in group T received periodontal therapy after the ligation. All the rats were sacrificed at the age of 24 weeks for measurement of blood lipids, insulin and blood glucose levels, and insulin resistance index (HOMA-IR) was calculated. The expressions of insulin receptor substrate-1 (IRS-1) and IRS-2 in the liver tissues were detected using real-time quantitative polymerase chain reaction (RT-PCR). RESULTS: Compared with the obese rats in group O, the rats in group P showed significantly higher HOMA-IR and LDL-C and lower expressions of IRS-1 and IRS-2 mRNA expression and HDL-C level (P<0.05). Compared with those in group P, the mRNA expressions of IRS-1 and IRS-2 and HDL-C level were significantly increased and LDL-C level, TC level and HOMA-IR were all decreased in group T (P<0.05), but the level of TG was comparable between the two groups. Pathological examination revealed lessened inflammatory cell infiltration and tissue destruction in the upper jaw of the rats in group T; the rats in group P presented with the most obvious upper jaw destruction and steatosis and inflammatory cell infiltration in the liver. CONCLUSION: Periodontal inflammation can downregulate the expression of IRS-1 and IRS-2 and increase insulin resistance and dyslipidemia in obese rats. Periodontal therapy produces a beneficial effect in improving insulin resistance and reducing dyslipidemia in obese rats.

[Effects of non-surgical periodontal therapy on serum inflammatory markers and metabolic level in obese rats].

OBJECTIVE: To investigate the effects of non-surgical periodontal therapy on serum inflammatory factors and metabolism levels in obese rats with experimental periodontitis. METHODS: Sixteen obese rats with experimental periodontitis were randomly divided into treatment group and control group with non-surgical periodontal therapy and no treatment, respectively. Oral glucose tolerance test was performed before treatment and 2 weeks after the treatment. All the rats were sacrificed 2 weeks after treatment and the orbital vein blood was taken to detect fasting blood glucose, fasting insulin, and serum level of C-reactive protein (CRP). Results Two weeks after periodontal treatment, fasting blood glucose (t=2.445, P=0.034) and beta cell function index (t=-2.543, P=0.027) were significantly lower in the treatment group than in the control group. Compared with those in the control group, CRP level (t=2.388, P=0.028) and the area under the curve in the oral glucose tolerance test (t=12.053, P=0.000) decreased significantly in the treatment group. CONCLUSION: Non-surgical periodontal treatment can reduce serum CRP level and improve glucose metabolism in obese rats.

[The effect of lead exposure in utero on the teeth eruption and enamel development of rat offspring.].

OBJECTIVE: To investigate the effects of lead exposure at different levels in utero on the teeth eruption and enamel development of rat offsprings. METHODS: 27 pregnant SD rats were divided into three groups randomly: high level lead group (HLG), low level lead group (LLG) and control group with nine rats in each group. The three groups from the gestation day to the end of the gestation were given either deionized water in control group or deionized water containing 200 mg/L Pb2+ as lead acetate through drinking method in high level lead experimental group and 50 mg/L Pb2+ as lead acetate through drinking method in low level lead experimental group. The incisors of newborn rats were marked at the level of the gingival papilla on the 26th day after birth. On the 36th day, the incisors of newborn rats were marked again at the same level. Then the rat offsprings were killed and their blood was collected for lead analysis. The mandible incisors of rat offspring were separated and the content of Pb in incisors was determined by using a graphite furnace atomic absorption spectrometric method. The teeth of rat offspring were observed and the distance between two marks were measured by means of stereomicroscope. The ratio of calcium to phosphate of enamel of rat offspring was compared by electron probe microanalyses. RESULTS: The level of blood lead in 200 mg/L, 50 mg/L treated rat offspring groups was higher than that in control group. The tooth lead of 200 mg/L, 50 mg/L treated rat offspring groups [(77.3 +/- 6.3), (27.8 +/- 4.5) microg/g] were higher than the control [(6.6 +/- 0.8) microg/g, P < 0.01]. Compared with the control group, the teeth of lead exposure experimental groups were smaller and severity of attrition was obvious and pulpal perforations were often observed. These appearances was more distinct in rats of high level lead experimental group. The incisors of lead-treated rat offspring erupted [(0.25 +/- 0.08), (0.30 +/- 0.09) mm/d] more slowly than control ones [(0.39 +/- 0.09) mm/d, P < 0.01]. The ratio of calcium to phosphate (Ca/P) decreased with the increase of lead exposure. It was found that Ca/P in lead exposure experimental groups (1.68 +/- 0.54), (1.37 +/- 0.47) was significantly lower than that in the control group (2.14 +/- 0.33). CONCLUSION: Lead exposure in utero affects the normal eruption of teeth and enamel formation and the degree was related with the lead exposure level.

[Construction of eukaryotic expression clone for human amelogenin].

OBJECTIVE: To construct-the eukaryotic expression clone for human amelogenin. METHODS: Total RNA was isolated from human fetal tooth buds. RT-PCR was used to amplify the amelogenin encoding region, and the amplified fragment for human amelogenin was inserted into eukaryotic expression vector PcDNA 3.1. The positive clones were selected and analyzed by restriction endonuclease mapping and DNA sequencing. RESULTS: 570 bp fragment was produced by RT-PCR; it was of the same size as expected based on human ameloginin mRNA encoding area length. The sequence of the inserted fragment from the recombinant clone PcDNA 3.1-AMG was consistent with that of AMELX from GenBank with one mismatch on 485 from G to C, without affecting the amino acid sequence. CONCLUSION: The eukaryotic expression clone PcDNA 3.1-AMG was successfully constructed with the properly inserted DNA sequence encoding mature human amelogenin.

[Attenuation of cardiac ischemia/reperfusion injury by transient low hydroperoxide pretreatment in rat].

OBJECTIVE: To investigate how transient low dose of hydroperoxide pretreatment prevents cardiac ischemia/reperfusion injury. METHODS: SD rats were divided into 4 groups: sham operation (Sham), standard ischemia/reperfusion (I/R), ischemic preconditioning (IPC) and IR preceded by low H2O2 treatment. Cardiac function and injury parameter were compared among groups. RESULTS: IPC protected reperfusion injury and improved cardiac function. Low H2O2 treatment played a role in cardioprotection similar to IPC. Low H2O2 was indeed generated in the early phase of simulated ischemia and attenuated cytochrome c release induced by high Ca2+ in isolated mitochondria. CONCLUSION: Low H2O2 plays a critical role in cardioprotection probably by inhibiting mitochondrial permeability transition.

[Meta analysis of association between TNF-α-308 polymorphism and periodontitis in Chinese Han population].

PURPOSE: To assess the association between TNF-α-308 polymorphism and periodontitis in Chinese Han population. METHODS: Articles published between 1994 and 2011 were searched in the fulltext database of CNKI, Chinese Biomedical Literature Database, Wanfang Database and PubMed. Case-control studies on TNF-α-308 polymorphism and periodontitis were searched up to Feb.,2011, including articles in Chinese and in English. Meta analysis was performed to evaluate the prevalence of Allele 2 between case group and control group. RESULTS: Five studies were entered into Meta analysis,with 494 cases and 501 controls . No significant heterogeneity was found among the studies(P=0.38); the pooled OR value of allele 2 was 2.12(95% CI 1.57-2.86), with significant difference(P<0.01). CONCLUSION: Meta analysis showed that TNF-α-308 polymorphism was associated with periodontitis in Chinese Han population.

[A national survey on dentin hypersensitivity in Chinese urban adults].

OBJECTIVE: To investigate the prevalence of dentin hypersensitivity in Chinese urban adults aged between 20 - 69 years old and the factors related to dentin hypersensitivity. METHODS: The Chinese national survey on dentin hypersensitivity was conducted in 20 - 69 years old adults in six representative cities, Beijing, Shanghai, Guangzhou, Wuhan, Chengdu, and Xi'an in 2008. A multi-stage stratified randomizing sampling method was used. Subjects were recruited from 36 urban survey sites in 6 cities. A structured questionnaire and a clinical examination on dentin hypersensitivity were used in the survey. The dentin hypersensitivity was diagnosed by a subject self-perceived short, sharp pain in response to a blast of cold air from a triple syringe administered to a tooth surface in 1 cm. RESULTS: In total, 7939 twenty to sixty-nine years old subjects completed a structured interview and underwent a clinical examination on dentin hypersensitivity. Among them, 40.7% (3230/7939) of the subjects reported being suffered from teeth sensitivity. When confirmed using a blast of air from a triple syringe and by ruling out other causes of sensitivity, such as caries, the prevalence was 29.7% (2354/7939), and the mean number of sensitive teeth was 1.4. The highest prevalence of dentin hypersensitivity [39.1% (622/1592)] was found in 50 - 59 years old group. The commonest teeth affected were the premolar teeth and the commonest initiating factor was cold drinks. Female, low education level, with gingival recession, attachment loss, and with the history of acidic substances derived from the stomach was related to dentin hypersensitivity. CONCLUSIONS: Dentin hypersensitivity was common in 20 - 69 years old Chinese urban adults. Dental professionals should give further emphasis to it.

[Differentiation characteristics of human periodontal ligament cell population in vitro].

OBJECTIVE: To explore the multi-differentiated capability of human periodontal ligament cell population (hPDLP), and provide a theoretical basis for the periodontal regeneration by tissue engineering technique. METHODS: hPDLP was cultured from periodontium of human tooth by the outgrowth method. STRO-1 and CD 146 expression were investigated by flow cytometry. hPDLP was induced to odontogenic/osteogenic-like and adipogenic-like cell. The multilineage differentiation capacities of hPDLP were evaluated by alizarin red stain, oil red O stain, anti-CD146 and STRO-1 immunocytochemistry, and reverse-transcriptase polymerase chain reaction analysis. RESULTS: hPDLP was isolated from human periodontium and most of the cells retained their fibroblastic spindle shape. hPDLP can be induced into osteoblast-like cells and adipocyte-like cells, and calcium deposition and lipid droplets were detected perspectively. And the eighth generation of hPDLP had weaker potential into adipocyte-like cells than the first passage, however, there was no difference to the aspect of calcification ability between the two passages. CONCLUSION: hPDLP cultured in vitro can differentiate into adipocytes and osteoblasts, and the first to third passage cells may have the predominance of differentiation potential.

[Expression and purification of human amelogenin mature peptide in Escherichia coli].

OBJECTIVE: To establish the expression and purification route for human amelogenin mature peptide in Escherichia coli and obtain the purified amelogenin (AMG) mature peptide. METHODS: Recombined plasmid pGEX-4T-1-AMG was transformed to Escherichia coli BL21. After expression, AMG was purified with glutathione S-transferase fusion protein purification system (GSTrapFF) column. RESULTS: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting hybridization results showed that 45,000 GST-AMG fusing protein and 19,000 target AMG mature peptide were obtained successfully. CONCLUSIONS: pGEX-4T-1-AMG-BL21 system is used successfully to express and purify human AMG mature peptide.

[Culture and identify the human umbilical vein endothelial cells and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains in the cells].

OBJECTIVE: To study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs. METHODS: HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry. RESULTS: HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%. CONCLUSION: Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.

[Construction of the eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 and green fluorescent protein].

OBJECTIVE: To construct an eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and enhanced green fluorescent protein (EGFP) controlled by cytomegalovirus promoter using pIRES-EGFP vector. METHODS: The mtHSP70 gene fragment was amplified by PCR from pVAX-mtHSP70-HSV2gD using specific primers. The PCR product was cloned into the vector pMD 18-T vector, and the correct clone was selected according to DNA sequence analysis. The interested mtHSP70 gene fragment was subcloned into pCMV-IRES-EGFP vector with XhoI and EcoR I digestion. The recombinant plasmid was transfected into mouse melanoma B16 cell line, and the green fluorescent cells were detected by fluorescence microscopy and mtHSP70 expression was detected by Western blotting. RESULTS: The recombinant plasmid obtained was confirmed by enzyme digestion. The transfected mouse melanoma B16 cells exhibited green fluorescence under fluorescence microscopy and expressed mtHSP70 protein as demonstrated by Western blotting. CONCLUSION: The eukaryotic coexpression vector PCMV-mtHSP70-IRES-EGFP has been established to allow further investigation of the role of mtHSP70 gene in tumor immunotherapy.

[Effects of lactational dioxin exposure to development of alveolar bone in SD rat offspring].

OBJECTIVE: To study the influence of lactational dioxin exposure (2, 3, 7, 8-tetrachlorodibenzo-p-dixon, TCDD) on development of alveolar bone in SD rat offspring. METHODS: The rats of TCDD exposure group and control group were sacrificed and the alveolar bone with molars of PD60 rats in the two groups were embedded in resin. The sections were observed by fluorescent microscope. The alveolar bone formation was evaluated by histological examination, tetracycline fluorescence marker and quantitative histomorphometry. The indices of quantitative histomorphometry were compared. RESULTS: The trabecular structure of alveolar bone was looser in TCDD exposure group than in the control group. The tetracycline fluorescence markers were more disorganized in TCDD group. The indices of quantitative histomorphometry of alveolar bone between two groups showed significantly different. CONCLUSIONS: Lactational 2,3,7,8-TCDD exposure decreased the quality and quantity of alveolar bone in SD rat offspring. It is suggested that dioxins exposure may interrupt the spatial configuration.

[Expression and purification of human amelogenin in Escherichia coli].

OBJECTIVE: To establish the expression and purification route for the gene encoding human amelongenin (AMG) mature peptide in Escherichia coli (E. coli). METHODS: Recombined plasmid pGEX-4T-1/AMG was identified by double endonuclease digestion electrophoretogram and DNA sequence analysis. The recombined plasmid was transformed to E. coli BL21. The inducing time, isopropyl-beta-D-thiogalactopyranoside (IPTG) concentration and inducing temperature were optimized for the express system. Under the optimized condition, the target fusing protein in superatant, periplasm, plasm and inclusion body was analyzed separately. A great amount of target fusing protein was found in the dissoluble protein. AMG fusing protein was purified by the GSTrapFF affinity column. RESULTS: Double endonuclease digestion electrophoretogram and DNA sequence analysis were done to identify the recombined vector pGEX-4T-1/AMG. The results were consistent with the anticipation. The optimum inducing time was 14.5 hours. The optimum IPTG concentration was 1.0 mmol/L. The optimum inducing temperature was 20 degrees C. Under this condition, the target protein was expressed to a maximum. Plentiful target protein was expressed in plasm and inclusion body under the optimized condition. A mount of plasm protein was obtained and purified by the GSTrapFF affinity column. The purified liquid was collected and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PACE). The protein electrophoresis map showed that AMG fusing protein was purified successfully. After twice elution, high pure fusing protein was obtained. CONCLUSION: pGEX-4T-1/AMG system is used successfully to express human AMG fusing protein.

[Study on the inhibition of vascular endothelial growth factor expression in Tca8113 cell by inducible nitric oxide synthase gene RNA interference].

OBJECTIVE: To investigate the effect of inducible nitric oxide synthase (iNOS) on vascular endothelial growth factor (VEGF) expression. METHODS: The vector containing short hairpin RNA of iNOS was transfected into Tca8113 cells using the RNA interference (RNAi) technique. The gene and protein expression of iNOS and VEGF was examined by RT-PCR and Western blot. RESULTS: The expression of iNOS, VEGF gene in Tca8113 cells was significantly different between the experimental and control groups 24 h and 48 h after transfection (P < 0.05). The protein expression of iNOS was different between the two groups 36 h and 48 h after transfection, and of VEGF was also different between the two groups 48 h after transfection (P < 0.05). CONCLUSIONS: The expression of VEGF could be down-regulated by silencing the iNOS gene in Tca8113 cells.