Efficacy and safety of salmeterol/fluticasone compared with montelukast alone (or add-on therapy to fluticasone) in the treatment of bronchial asthma in children and adolescents: a systematic review and meta-analysis / 中华医学杂志(英文版)

BACKGROUND@#Despite the recommendation of inhaled corticosteroids (ICSs) plus long-acting beta 2-agonist (LABA) and leukotriene receptor antagonist (LTRA) or ICS/LTRA as stepwise approaches in asthmatic children, there is a lack of published systematic review comparing the efficacy and safety of the two therapies in children and adolescents aged 4 to 18 years. This study aimed to compare the safety and efficacy of salmeterol/fluticasone (SFC) vs. montelukast (MON), or combination of montelukast and fluticasone (MFC) in children and adolescents aged 4 to 18 years with bronchial asthma.@*METHODS@#A systematic search was conducted in MEDLINE, EMBASE, the Cochrane Library, China BioMedical Literature Database, Chinese National Knowledge Infrastructure, VIP Database for Chinese Technical Periodical, and Wanfang for randomized controlled trials (RCTs) published from inception to May 24, 2021. Interventions are as follows: SFC vs. MON, or combination of MFC, with no limitation of dosage or duration. Primary and secondary outcome measures were as follows: the primary outcome of interest was the risk of asthma exacerbation. Secondary outcomes included risk of hospitalization, pulmonary function, asthma control level, quality of life, and adverse events (AEs). A random-effects (I2 ≥ 50%) or fixed-effects model (I2 < 50%) was used to calculate pooled effect estimates, comparing the outcomes between the intervention and control groups where feasible.@*RESULTS@#Of the 1006 articles identified, 21 studies met the inclusion criteria with 2643 individuals; two were at low risk of bias. As no primary outcomes were similar after an identical treatment duration in the included studies, meta-analysis could not be performed. However, more studies favored SFC, instead of MON, owing to a lower risk of asthma exacerbation in the SFC group. As for secondary outcome, SFC showed a significant improvement of peak expiratory flow (PEF)%pred after 4 weeks compared with MFC (mean difference [MD]: 5.45; 95% confidence interval [CI]: 1.57-9.34; I2 = 95%; P = 0.006). As for asthma control level, SFC also showed a higher full-controlled level (risk ratio [RR]: 1.51; 95% CI: 1.24-1.85; I2 = 0; P < 0.001) and higher childhood asthma control test score after 4 weeks of treatment (MD: 2.30; 95% CI: 1.39-3.21; I2 = 72%; P < 0.001) compared with MFC.@*CONCLUSIONS@#SFC may be more effective than MFC for the treatment of asthma in children and adolescents, especially in improving asthma control level. However, there is insufficient evidence to make firm conclusive statements on the use of SFC or MON in children and adolescents aged 4 to 18 years with asthma. Further research is needed, particularly a combination of good-quality long-term prospective studies and well-designed RCTs.@*PROSPERO REGISTRATION NUMBER@#CRD42019133156.

1,25-(OH)2D3 inhibits lipopolysaccharide-induced expression of interleukin-13 and interleukin-17 in cord blood CD4+T cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of 1,25-(OH)2D3 on lipopolysaccharide (LPS)-induced expression of interleukin-13 (IL-13) and interleukin-17 (IL-17) in cord blood CD4(+)T cells, providing theoretical basis for clinical reasonable application of vitamin D and prevention of asthma and allergic diseases.</p><p><b>METHODS</b>Mononuclear cells (MNCs) were isolated from umbilical cord blood (50 mL) of 12 normal eutocia term newborns by gravity centrifugation. CD4(+)T cells were isolated using magnetic beads, which was cultured with following three kinds of stimulus for 72 hours: natural state (blank group), LPS (10 μg/mL)stimulation alone and LPS(10 μg/mL)+1,25-(OH)2D3 (10(-8) mmol/L)stimulation. Levels of IL-13 and IL-17 in the culture supernatant and mRNA expressions in cord blood CD4(+)T cells were detected using ELISA and real Time-PCR respectively.</p><p><b>RESULTS</b>Compared with the blank group, levels of IL-13 and IL-17 in the culture supernatant and mRNA expression of IL-13 and IL-17 in the cord blood CD4(+)T cells increased in the LPS stimulation alone group (P<0.01). When co-stimulation of 1,25-(OH)2D3 with LPS, levels of IL-13 and IL-17 in the culture supernatant and mRNA expression of IL-13 and IL-17 in the cord blood CD4(+)T cells decreased compared with LPS-stimulated alone group (P<0.05), but remained higher than the blank group (P<0.01).</p><p><b>CONCLUSIONS</b>LPS can promote expression of IL-13 and IL-17 in cord blood CD4(+)T cells. 1,25-(OH)2D3 inhibits the expression of IL-13 and IL-17, suggesting that vitamin D intake may provide protective effects in the development of atopy-predisposing immune responses in early life.</p>

Effect of 1,25-(OH)2D3 supplementation during gestation and lactation on TGF-β1 and Smad3 expression in lungs of rat offspring with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of 1,25-(OH)2D3 supplementation during gestation and lactation on TGF-β1 and Smad3 expression in lungs of rat offspring with asthma.</p><p><b>METHODS</b>Thirty-two female Wistar rats were randomly divided into four groups: low-, medium- and high-dose 1,25-(OH)2D3 supplementation and control groups (n=8 each). From the 7th day of gestation, the three 1,25-(OH)2D3 supplementation groups were administered with 2,10 and 20 μg/mL of 1,25-(OH)2D3 respectively every other day until weaning (rat offspring: 21 days old). The control group received normal saline instead. Then, bronchial asthma was induced in rat offspring from the 4 groups. The protein and mRNA expression of TGF-β1 and Smad3 in the lung tissue was measured by immunochemistry and RT-PCR.</p><p><b>RESULTS</b>Eosinophil cell infiltration and airway inflammation decreased in rat offspring from the low- and medium-dose 1,25-(OH)2D3 groups, but increased in rat offspring of the high-dose 1,25-(OH)2D3 group compared with the control group. Immunohistochemistry of lung tissues showed that the expression of TGF-β1 protein and pSmad3 decreased in rat offspring from the low- and medium-dose 1,25-(OH)2D3 groups (P<0.05), but increased significantly in rat offspring from the high-dose 1,25-(OH)2D3 group compared with the control group (P<0.05). PCR showed that the expression of TGF-β1 and Smad3 mRNA in the lung tissue decreased in rat offspring from the low- and medium-dose 1,25-(OH)2D3 groups (P<0.05), but increased significantly in rat offspring from the high-dose 1,25-(OH)2D3 group compared with the control group (P<0.05).</p><p><b>CONCLUSIONS</b>1,25-(OH)2D3 supplementation plays a role in regulating the immune system in asthmatic rats. Its mechanism may be associated with regulation of the expression of TGF-β/Smad signal pathway-related proteins through the vitamin D receptor signal pathway.</p>

Effect of montelukast sodium on TGF-beta(1) of peripheral blood mononuclear cells from children with mild persistent asthma / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the role of transforming growth factor beta(1) (TGF-beta(1)) in the pathogenesis of bronchial asthma in children and assess the effect of montelukast sodium (leukotriene receptor antagonist) on TGF-beta(1) levels.</p><p><b>METHOD</b>A 12 weeks single-blind, placebo-controlled trail was conducted in 60 children with mild persistent asthma [aged 5 - 14 years, mean (7.10 ± 0.27) years]. Patients were randomly assigned to receive 5 mg montelukast sodium or placebo for 12 weeks. And 30 healthy control children [aged 5 - 14 years, mean (7.60 ± 0.25) years] were also recruited in this study from Sep. 2009 to Sep. 2010. Clinical effects and pulmonary function were evaluated before and 12 weeks after treatment. The mRNA expression of TGF-beta(1) in the peripheral blood mononuclear cells was detected by using RT-PCR with beta-actin as internal control. The percentage of the different subpopulations of Foxp(3)(+)CD4(+) T cells was assayed by 4-color flow cytometric analysis system and the levels of TGF-beta(1) in plasma by ELISA.</p><p><b>RESULT</b>(1) The basic characteristics between asthma group and healthy group had no significant difference. (2) Following treatment, there was significant increase in pulmonary function in asthmatic children. The effect in the group of montelukast sodium was superior to that of placebo group (P < 0.05). (3) The serum expression of TGF-beta(1) in asthmatic children was lower than that in control group (q = 20.01, P < 0.01); after 12 weeks of treatment, the mean expression of TGF-beta(1) was (20.03 ± 1.14) ng/L for montelukast sodium group and (12.10 ± 3.91) ng/L for placebo group (P < 0.05). (4) The mRNA expression of TGF-beta(1) in asthma children was lower than that in control group (0.31 ± 0.07 vs 0.61 ± 0.2, q = 8.97, P < 0.05); after 12 weeks of treatment, the mean expression of TGF-beta(1) was (0.46 ± 0.13) for montelukast sodium group and (0.32 ± 0.04) for placebo group (q = 8.25, P < 0.05). (5) It was shown that the total Foxp(3)(+)CD(4)(+) cell percentage was higher in asthmatic children than those of control group (8.30% ± 1.30% vs 6.05% ± 1.80%); the proportion of the three subpopulation was different between groups: CD(45) RA(+)Foxp(3)(lo) was higher in asthmatic group (4.60% ± 1.04% vs 3.27% ± 1.03%) and CD(45) RA(-)Foxp(3)(hi) was lower (0.75% ± 0.13% vs 0.93% ± 0.26%); while CD(45) RA(-)Foxp(3)(lo) had no significant difference among groups (2.40% ± 0.83%, 1.61% ± 1.10%). After 12 weeks of treatment, the percentage of CD(45) RA(-)Foxp(3)(hi) was increased in montelukast sodium group compared with placebo group (1.16% ± 0.24% vs 0.89% ± 0.22%). (6) Spearman correlation analysis revealed that TGF-beta(1) levels had no correlation with the levels of pulmonary function.</p><p><b>CONCLUSION</b>The protein and mRNA expression level of TGF-beta(1) was low in those asthmatic children. Insufficient secretion of TGF-beta(1) and the defective ability of activated regulatory T cells (CD(45) RA(-)Foxp(3)(hi)) in Foxp(3)(+)CD(4)(+) Treg cells might play an important role in pathogenesis of asthma. Up-regulation of the expression of TGF-beta(1) and induction of the expression of CD(45) RA(-)Foxp(3)(hi) in Foxp(3)(+)CD(4)(+)Treg cells by montelukast sodium may be one of the immunomodulatory mechanisms in asthma.</p>

Inhibition effects of tumor necrosis factor-alpha gene transduced tumor drainage node of lymphocytes from tongue cancer on SCID mice transplanted tumor established with human tongue carcinoma cell lines / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the in vivo inhibition effects of tumor necrosis factor-alpha (TNF-alpha) gene transduced tumor drainage node of lymphocytes (DNL) from tongue cancer on SCID mice transplanted tumor.</p><p><b>METHODS</b>15 human tongue carcinoma models were established in SCID mice by subcutaneously injection of squamous cell carcinoma line Tca8113. TNF-alpha gene introduced DNL, combined with low dose Pinyancin (PYC), were locally injected into tumor site. The inhibition rate was determined by the weights at the 8th week after tumor dissection and fresh specimens were prepared and subject to histopathologic examination under transmission electron microscope, and in situ TUNEL was used to detect apoptosis.</p><p><b>RESULTS</b>The TNF/DNL and rIL-2 group, and the TNF/DNL and rIL-2 and PYC group both exerted a strong inhibition effect on the implanted tumor. Treated tumors of the TNF/DNL and rIL-2 and PYC group were significantly reduced in comparison with those of the TNF/DNL and rIL-2 group (P < 0.05). The apoptosis of tumor in the TNF/DNL and rIL -2 group was evidenced based on transmission electron microscope and TUNEL analysis, and the apoptosis index was higher than that of control group (P < 0.05).</p><p><b>CONCLUSION</b>Local injection of DNL modified with TNF-alpha gene, combined with low dose PYC, exert a synergistic antitumor effect. Apoptosis may be an important mechanism of squamous cell carcinoma killed by TNF/DNL.</p>

Effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells.</p><p><b>METHODS</b>Oral leukoplakia cell line (Leuk-1) transfected with pCMV-Podoplanin (A4-1) or pCMV (B4-1) was used in this study. The levels of mRNA and protein of Podoplanin were detected by real-time PCR and Western boltting in A4-1, B4-1 and Leuk-1 cells. The localization of Podoplanin was detected by confocal microscope. Cell growth and proliferation was examined by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry (FCM).</p><p><b>RESULTS</b>The value of Podoplanin mRNA level in A4-1 cells was 0.022, which was significantly higher than the values in B4-1 (0.001) and Leuk-1 cells (0.002), P < 0.05. The gray scale of Podoplanin protein in A4-1 cells was significantly higher than in the control groups (P < 0.05). The expression of Podoplanin was observed in cell plasm and membrane of A4-1, B4-1 and Leuk-1 cells. But the expression level of Podoplanin in A4-1 cells was higher than in control groups. A4-1 cells grew faster than Leuk-1 cells. The proliferation rate after 3 days of culture in A4-1 cells was 40.4% higher than that in B4-1 cells (P < 0.05). The G₂-M phase (24.89 +/- 4.55)% and PI (0.57 +/- 0.06) of A4-1 cells were significantly higher than that in B4-1 cells [(4.13 +/- 5.24)%, (0.41 +/- 0.04)] and Leuk-1 cells [(4.69 +/- 7.42)%, (0.40 +/- 0.02)], P < 0.05.</p><p><b>CONCLUSIONS</b>Over expression of Podoplanin accelerated the growth and proliferation of oral leukoplakia cells. Podoplanin may be one of key genes in the malignant transformation of oral leukoplakia.</p>

Inflammatory factors promote oral squamous cell carcinoma cells metastasis, via nuclear factor kappa B signal pathway in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the roles of inflammatory factors and nuclear factor kappa B (NF-kappaB) signal pathway in metastasis of oral squamous cell carcinoma.</p><p><b>METHODS</b>The oral squamous cell carcinoma cell lines with highly metastasis potential (Tb) and lower metastasis potential (Tca8113) were used in this study. The levels of NF-kappaB activity in oral squamous cell carcinoma cell lines were determined by Western blotting and luciferase reporter assay. pBalphabe-IkappaBalpha-SR expression vector or NF-kappaB inhibitor pyrolidinedithiocarbamate (PDTC) was used to inhibit NF-kappaB, and cell migration was examined by transwell assay. The secretion of tumor necrosis factor-alpha (TNF-alpha), IL-1alpha, IL-6, IL-8 and GM-CSF proinflammatory cytokines was determined by ELISA when Tb cells were transfected with pBalphabe-SR-IkappaBalpha or treated with PDTC.</p><p><b>RESULTS</b>Western blotting showed that the levels of phosphorIkappaBalpha and phosphor-p65 were highly expressed in Tb cells. Tb cells had high level of constitutive NF-kappaB activity and were more sensitive to TNF-alpha. The migration of highly metastatic Tb cells, either transfected with dominant-negative mutant inhibitor pBalphabe-SR-IkappaBalpha or treated with PDTC, was suppressed when determined by transwell assay. The secretion of proinflammatory cytokines, including TNF-alpha, IL-1alpha, IL-6, IL-8 and granulocyte-macrophage colony stimulating factor (GM-CSF), was inhibited by pBalphabe-SR-IkappaBalpha transfection or PDTC treatment.</p><p><b>CONCLUSIONS</b>The inflammatory factors such as TNF-alpha could promote oral squamous cell carcinoma cell metastasis via NF-kappaB signal pathway.</p>

Effect of vitamin D supplementation in early life on the expression of interleukin-10 and intercellular adhesion molecule-1 in rat asthma model / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the effect and mechanism of vitamin D supplementation in early life on rat asthma model.</p><p><b>METHOD</b>Thirty two sex-mature, female Wistar rats were randomly divided into a control group (n = 8), a low dose group (n = 8), a medium dose group (n = 8) and a high dose group (n = 8). From the seventh day of pregnancy on, the rats in each group were given different doses of vitamin D by intragastric administration, until the offspring rats were 21 days old. The rats in the control group were fed with DMSO-PBS. After the offsprings were weaned, 8 rats were randomly selected from each group. The number of male and female rats was equal. The rats were sensitized to ovalbumin (OVA) and challenged with aerosol OVA to establish the asthma model. The lung tissues were examined for pathologic changes after HE staining. ELISA was used to determine the concentrations of IL-10 in serum and BALF. Immunohistochemical staining methods were used to measure the expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissues.</p><p><b>RESULT</b>(1) Pathologic changes of lung tissues: compared with the control group, light microscope (LM) showed that eosinophil cells infiltration and the airway inflammation decreased in the low dose and medium dose groups, but increased in the high dose group. (2) The concentrations of IL-10 in serum and BALF: In serum, compared with the control group [(18.7 +/- 4.7) pg/ml], the concentrations of IL-10 in the low dose group [(30.2 +/- 2.8) pg/ml, P < 0.05] and the medium dose group [(51.5 +/- 6.6) pg/ml, P < 0.05] were significantly increased. And the IL-10 level of medium dose group was higher than that of the low dose group (P < 0.05). In BALF, compared with the control group [(59.1 +/- 14.4) pg/ml], the concentrations of IL-10 in the medium dose group [(90.0 +/- 14.3) pg/ml, P < 0.05] was significantly increased. There were no significant changes in the low dose group [(58.1 +/- 3.4) pg/ml, P > 0.05], whereas in the high dose group [(45.3 +/- 6.5) pg/ml, P < 0.05] the level significantly decreased. (2) The expression of ICAM-1 in lung tissues: compared with the control group, there were no significant changes in the low dose group (P > 0.05). The expression of ICAM-1 was significantly decreased in the medium dose group (P < 0.05). In the high dose group, the expression of ICAM-1 was significantly increased (P > 0.05).</p><p><b>CONCLUSION</b>Adequate intervention with 1,25(OH)2D3 in the early life could alleviate the inflammation in the lung tissues, reduces eosinophil cell infiltration in rat asthma model. However, overdose might play a detrimental role. Its mechanism may be associated with the effect of 1,25(OH)2D3 on IL-10 secretion and the expression of ICAM-1.</p>

Benzo (a) pyrene induced tumorigenesity of human immortalized oral epithelial cells: transcription profiling / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The present study was designed to examine and analyze the global gene expression changes during the tumorigenesis of a human immortalized oral epithelial cell line, and search for the possible genes that may play a role in the carcinogenesis of oral cancer associated with benzo (a) pyrene.</p><p><b>METHODS</b>The human immortalized oral epithelial cells, which have been established through transfection of E6/E7 genes of human papillomavirus type 16 and proved to be non-tumorigenic in nude mice, were treated with benzo (a) pyrene. Tumorigenicity of the treated cells were examined through nude mice subcutaneous injection. The global gene expression profiles of immortalized cells and the tumorigenic cells were acquired through hybridization of a microarray of Affymetrix U133 plus 2.0. The data were analyzed using Spring 7.0 software and treated statistically using one-way analysis of variance (ANOVA). The differentially expressed genes were classified using a Venn diagram and annotated with gene ontology. Several highlighted genes were validated in cells using a real-time polymerase chain reaction.</p><p><b>RESULTS</b>There were 883 differentially expressed genes during the tumorigenesis and most of them changed expression in the early stage of tumorigenesis. These genes mainly involved in macromolecule metabolism and signal transduction, possessed the molecular function of transition metal ion binding, nucleotide binding and kinase activity; their protein products were mainly integral to membranes or localized in the nucleus and cytoskeleton. The expression patterns of IGFBP3, S100A8, MAP2K, KRT6B, GDF15, MET were validated in cells using a real-time polymerase chain reaction; the expression of IGFBP3 was further validated in clinical oral cancer specimens.</p><p><b>CONCLUSIONS</b>This study provides the global transcription profiling associated with the tumorigenesis of oral epithelial cells exposed to benzo (a) pyrene; IGFBP3 may play a potential role in the initiation of oral cancer related to benzo (a) pyrene exposure.</p>

DNA methylation in salivary adenoid cystic carcinoma cell lines / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the promoter methylation status of tumor suppressor gene and the relationship between promoter methylation and mRNA, protein expression of tumor suppressor gene in salivary adenoid cystic carcinoma (ACC) cell lines.</p><p><b>METHODS</b>Promoter methylation status of E-cad, p16, RASSF1A, DAPK, and MGMT was determined by methylation-specific polymerase chain reaction (MSP) in ACC cell lines, ACC-2, ACC-3, and ACC-M. E-cad, p16 protein and mRNA expression was also examined by IHC and RT-PCR in 3 ACC cell lines.</p><p><b>RESULTS</b>All the three salivary ACC cell lines exhibited E-cad, p16 promoter methylation, but no methylation of RASSF1A, DAPK, and MGMT was found. There was p16 protein and mRNA expression but no E-cad expression in 3 ACC cell lines.</p><p><b>CONCLUSIONS</b>The results suggest that in ACC cell lines, promoter methylation of E-cad, p16 is a common event, and promoter methylation may be one of the major mechanism for inactivation of E-cad.</p>

Synthesized small interfering RNA inhibits the expression of DNA methyltransferase 1 in salivary adenoid cystic carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To select the effective siRNA which could inhibit the expression of DNA methyltransferase 1 (DNMT-1) in adenoid cystic carcinoma (ACC) and discuss the time-, and dose-dependent effect of RNA interference (RNAi).</p><p><b>METHODS</b>Four pairs of siRNA were designed, synthesized and transfected through oligofectamine reagent into ACC cell lines ACC-2, ACC-3 and ACC-M. 24, 48 and 72 h after transfection, total RNA and protein were harvested respectively. mRNA and protein expression level of DNMT-1 were detected by real time PCR, RT-PCR and Western blot, and then the effective siRNA was subsequently selected. ACC-3 as also transfected by different concentration of siRNA and the dose-dependent effect of RNAi was discussed. Cy(3) labelled GAPDH siRNA was used as a positive control.</p><p><b>RESULTS</b>Two of 4 pairs of siRNA inhibited the mRNA expression of DNMT-1 in three ACC cell lines and the expression of DNMT-1 was downregulated by 67%, 86%, 92% and 76%, 76%, 86% respectively. The gene inhibition was detected at 24 h or 48 h after transfection, maintained only 1 - 2 days and showed direct relationship to the concentration of siRNA. The change of protein expression level of DNMT-1 was consistent to the changes of mRNA.</p><p><b>CONCLUSIONS</b>The effective siRNA which could inhibited the expression of DNMT-1 of ACC were achieved. The inhibition effect of RNAi was time-dependent and dose-dependent.</p>

Establishment and identification of biological characteristics of rat monoclone cell line Rca-B / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To establish a monoclone cell line of squamous cell carcinoma (SCC) in rat buccal mucosa and to study its biological characteristics.</p><p><b>METHODS</b>SCC in rat oral mucosa was induced by adding 4-nitroquinoline-1-oxide (4NQO) into the SD rats' drinking water, and the cancer cells were then cultured to obtain mixed cells in vitro. The mixed tumor cells were purified by mono cell cloning method. The biological characteristics of the cells were studied by microscope and electronic microscope observation, chromosome analysis, Methyl thiazolyl tetrazolium (MTT) test, flow cytometry assay and immunohistochemistry staining. Hypodermic inoculations of the cells in nude mice and injection of the cells by nude mice tail veins were performed to observe the tumor formation and long distance metastasis.</p><p><b>RESULTS</b>The morphology proved that the cell line was squamous cell carcinoma cells, which were cultured from one cell. The population doubling time for passage 65 cells was 25.44 hours. The cells in S-phase accounted for 20.13% of the cell cycle. The chromosome modal number was 84. All the cells expressed the proteins of cytokeratin and vimentin. The xenograft rate and the tumor metastatic rate to the lung were 100% in nu/nu BALB/C mice, but the homograft rate was zero in SD Rats.</p><p><b>CONCLUSIONS</b>Rca-B was a typical oral squamous cell carcinoma cell line derived from Sprague-Dawley rat buccal mucosa carcinoma, and the cell line has high metastatic potential and its biological characteristics were well ascertained.</p>

Study on the pharmacokinetic profile of telbivudine / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the pharmacokinetic profile of telbivudine in healthy Chinese subjects after oral administration of single and multiple doses.</p><p><b>METHODS</b>Forty-two healthy adult male and female subjects 18-40 years of age were randomized into four telbivudine dosing groups of 200 mg, 400 mg, 600 mg and 800 mg. Subjects in the 600 mg group received both a single dose and once daily multiple doses for 8 consecutive days. Telbivudine concentrations in plasma and urine samples collected at different time points before and after the drug administration were measured using HPLC-MS/MS. Pharmacokinetic parameters were calculated by using the non-compartmental approach.</p><p><b>RESULTS</b>After a single dose of 200 mg, 400 mg, 600 mg and 800 mg, tmax (median) were 2.50, 2.00, 2.00 and 2.50 hours respectively; t1/2 were (43.3 +/- 15.2) h, (49.1 +/- 14.4) h, (39.4 +/- 12.1) h and (46.7 +/- 20.8) h respectively; Cmax were (1,753.2 +/- 389.0) ng/ml, (2,586.7 +/- 871.4) ng/ml, (3,703.6 +/- 1,219.0) ng/ml and (3454.6 +/- 953.9) ng/ml respectively; AUC(0-infinity) were (12,843.2 +/- 2,925.6) ng.h(-1).ml(-1), (22,948.9 +/- 5,721.0) ng.h(-1)/ml(-1), (26,440.5 +/- 8,938.1) ng.h(-1).ml(-1) and (28, 820.9 +/- 7 912.9) ng.h(-1).ml(-1) respectively, and CL(R) (600 mg) was (6,545.6 +/- 1 504.4) ml/h. The AUCss from multiple doses was (1,088.5 +/- 299.8) ng/ml; Cmax and AUC accumulation ratio were 1.02 +/- 0.21 and 1.23 +/- 0.26 respectively, which implicated moderated accumulation.</p><p><b>CONCLUSION</b>Pharmacokinetic parameters of telbivudine in Chinese healthy subjects were determined.</p>

The establishment of a carcinogenesis model of oral squamous cell carcinoma in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To transform HPV E6/E7 immortalized human oral epithelial cell (HIOEC) line cells by benzo(a)pyrene [B(a)P] in vitro, and to establish a carcinogenesis model of oral squamous cell carcinoma.</p><p><b>METHODS</b>HIOEC cells were treated with 0.1 mg/L-1.2 mg/L B(a)P for 6 months. The cells were cloned at 18th passage, and then the culture medium was changed into DMEM containing 10% FBS at 21th passage. Cells were cultured in vitro for half and one year and the cell line was named HIOEC-B(a)P. The morphological changes of the cells were observed with differential interference contrast microscope and HE staining. The soft agar colony forming ability and tumorigenicity of the cells in nude mice were identified to confirm the malignant characteristics of HIOEC-B(a)P cells.</p><p><b>RESULTS</b>(1) After HIOEC cells were treated with B(a)P for 6 months, HIOEC-B(a)P cells could grow well in DMEM medium containing 10% FBS and physical concentration of calcium. (2) When HIOEC cells were treated with chemical carcinogens, the morphology of the cells was changed. Cells showed the character of polygon epithelial cells with much atypical mitosis. (3) The 93th passage of HIOEC-B(a)P cells had soft agar colony formation ability. (4) The 55th passage of HIOEC-B(a)P cells could develop parakeratosis mass. The 69th passage of HIOEC-B(a)P cells could develop typical well-differentiated squamous cell carcinoma. The 74th and the 96th HIOEC-B(a)P cells developed I-II grade squamous cell carcinoma-like clinical lesions in nude mice.</p><p><b>CONCLUSIONS</b>B(a)P may induce HIOEC cells to be oral squamous cell carcinoma (OSCC) carcinogenetic cells. It will provide a multiple factors, multistage carcinogenesis model of OSCC for the further research.</p>

Experimental study of hydroxycamptothecine and theprubicine in the treatment of adenoid cystic carcinoma lung metastases / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate anticancer effect of hydroxycamptothecine and theprubicine in the treatment of established human adenoid cystic carcinoma (ACC) lung metastasis of nude mice and to determine the best therapy sequence of the two drugs.</p><p><b>METHODS</b>The model of ACC lung metastasis (60 animals) was established by injecting the ACC-M cells into the nude mice from their tail veins. Animals were divided into six groups: v acuity control group, HCPT group, THP group, HCPT and THP group 1, HCPT and THP group 2, HCPT and THP group 3 (No.1 - No.6). Different treatment sequence was performed on these groups. The lung samples were collected and observed after HE staining. The area of metastases and lung tissue were analyzed by the images analysis system.</p><p><b>RESULTS</b>The size of the metastatic lesion in the treatment groups were smaller than that in the control group (P < 0.01). The treatment effects between the group with simultaneous administration of HCPT and THP and the group with single drug administration were not different (P > 0.05). No.5 and No.6 groups showed better effect of treatment than the other treatment groups (P < 0.05).</p><p><b>CONCLUSIONS</b>There are two recommended sequences: (1) HCPT should be used after THP; (2) THP is used 48 hours, After HCPT is administered. Because using two drugs together will decrease the anticancer effect, HCPT and THP should not be used simultaneously.</p>

Inhibition effect on Tca8113/CDDP by antisense oligonucleotides of cyclin D1 combined with cis diamminedichloroplatinum / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To clarify the relationship between cyclin D1 and cisplatin resistance of Tca8113/cis diamminedichloroplatinum (CDDP) in vitro and in vivo.</p><p><b>METHODS</b>We applied the transfection method with plasmids pcDNA3.1-antisense-cyclin D1 by Lipofectamine 2000. Tca8113/CDDP cells were used as control. MTT assay was used to identify the proliferation and sensibility of those cells to cisplatin. Subsequently, 18 nude mice were subcutaneously injected by those cells and divided into 3 groups with 6 mice in each group. Every mouse was treated by cisplatin with 5 mg . kg(-1) . d(-1) for 5 days. The sizes of tumor were measured every other day and were described with the growth curves. After 20 days, tumors were anatomized and weighed. The tumor inhibition ratios were calculated and the HE slides were observed to determine the cell sensibility to cisplatin.</p><p><b>RESULTS</b>The transfected cells with pcDNA3.1-antisense-cyclin D1grew more slowly than other cells and showed higher sensibility to cisplatin in vitro. The tumors developed by cells with pcDNA3.1-antisense-cyclin D1 were smaller than the The tumor inhibition ratio was 74% (P < 0.05). The necrosis area was larger in the tumors developed by the transfected cells with pcDNA3.1-antisense-cyclin D1 than other groups.</p><p><b>CONCLUSIONS</b>Antisense oligonucleotides of cyclin D1 can improve the sensibility of Tca8113/CDDP cells to cisplatin and inhibit the growth of tumors.</p>

Effect of Yishenqinghuo compound on biological characteristics of rat bone marrow stromal cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of Yishenqinghuo recipe on biological characteristic of rat bone marrow stromal cells (BMSCs) and search for the function and mechanism of this recipe in treatment of periodontitis.</p><p><b>METHODS</b>12- to 15-month-old SD rats were allocated into 5 groups. Group A: control group (with no periodontitis model, fed with the same dosage of saline as Group D); Group B: model group (with periodontitis model, fed with the same dosage of saline as Group D); Group C: high dosage group (with periodontitis model, fed with a high dosage of medicine); Group D: middle dosage group (with periodontitis model, fed with a middle dosage of medicine); and Group E: low dosage group (with periodontitis model, fed with a low dosage of medicine). Ten days later, serums were collected from all the five groups for in vitro cultivation of BMSCs.</p><p><b>RESULTS</b>There was displayed a similarity in effect between the serum collected from Group C1 (serum collected 0.5 h after the last gavage of Group C) and that from Group A after the last gavage, the effect being the best in terms of proliferation of BMSCs. A comparison with the other groups had revealed a striking difference (P < 0.01), with Group B having turned out to be the worst. The serum from Group C2 (serum collected 1 h after the last gavage of Group C) after the last gavage could best enhance the generation and activity of ALP, having demonstrated a significant difference (P < 0.01) in comparison with the other groups.</p><p><b>CONCLUSION</b>Yishenqinghuo recipe can improve the proliferation of BMSCs and facilitate the differentiation to osteoblast.</p>

In vitro gene silencing by siRNA of cyclin D1 on tongue squamous cell carcinoma cell line resistant to cisplatin / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To interfere in the Tca8113-CDDP cell line with siRNA of cyclin D1 and to investigate time and dose dependent gene silencing effect of siRNA of cyclin D1.</p><p><b>METHODS</b>siRNA of cyclin D1 was transfected into Tca8113-CDDP cells Fluorescent CY3 dye labeled siRNA GAPDH was used as the control. The transient transfecting efficiency was examined at 4, 24, 48 and 72 h. The relative quantity of the target RNA of cyclin D1 was analyzed with SYBR Green fluorescent dye kit by the Real-time PCR assay. The protein level of cyclin D1 was examined with Western blot. The changes of cisplatin sensitivity after treatment with siRNA cyclin D1 were examined with methyl thiazolyl tetrazolium (MTT) assay.</p><p><b>RESULTS</b>The optimized transfecting efficiency with CY3 labeled siRNA GAPDH in Tca8113-CDDP cells was over 90%. The silencing rate of cyclin D1 siRNA was 81.6% at 24 h, 80.7% at 48 h and 94.3% at 72 h. Dose-dependent manner of gene silencing effect was observed when the siRNA concentration was lower than 100 nmol/L, however, gene silencing effect reached its platform when the concentration was higher than 100 nmol/L. The protein levels of cyclin D1 at 24, 48 and 72 h after transfection decreased significantly, and so did the growth of cells. Inhibition rates of cell growth induced by cisplatin after administration with or without cyclin D1 siRNA were 58.4% and 34.8%, respectively.</p><p><b>CONCLUSIONS</b>Chemical synthesized cyclin D1 siRNA effectively silenced the expression of cyclin D1 gene in Tca8113-CDDP cells in vitro, with a time- and dose-dependent manner and target gene silence in cells increased its sensitivity to cisplatin.</p>

The effect of intraperitoneal heparin on distribution of ACC-M in lung and liver / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of intraperitoneal heparin on the adhesion of highly lung metastasic adenoid cystic carcinoma cell line (ACC-M) in lung.</p><p><b>METHODS</b>3HTdR labeled ACC-M cells were injected by intravenous infusion after intraperitoneal injection with 200 units heparin. 4 mice of each group were killed at 2 h, 6 h and 18 h after infusion. The relative radioactivity (CPM) in lung and liver was detected.</p><p><b>RESULTS</b>3H-activity per gram in lung of heparin group was lower than control at the same time. There was significant difference between the two groups (P < 0.001). There was no difference between the two groups in liver (P > 0.05).</p><p><b>CONCLUSION</b>The results of this study suggest that the highly lung metastasis characteristic of ACC-M may be partially inhabited by the use of intraperitoneal heparin.</p>

Significance of C-Reactive Protein Monitoring to Guide the Course of Treatment with Antibiotic in Neonatal Bactenal Infection / 实用儿科临床杂志

0.05). There was only 2/109 cases (5.8%) need a second course of antibiotics because of likely infection and 102/109 cases (93.5%)need not any moor antibiotics. The mean period of antibiotic treatment in group Ⅰ, group Ⅱa and group Ⅱb were (1.2?0.5) days,(4.8?0.8) days and (9.3?1.8) days,respectively.There were significant differences(all P