Adult dental epithelial stem cell-derived organoids deposit hydroxylapatite biomineral / 国际口腔科学杂志·英文版

Ameloblasts are specialized cells derived from the dental epithelium that produce enamel, a hierarchically structured tissue comprised of highly elongated hydroxylapatite (OHAp) crystallites. The unique function of the epithelial cells synthesizing crystallites and assembling them in a mechanically robust structure is not fully elucidated yet, partly due to limitations with in vitro experimental models. Herein, we demonstrate the ability to generate mineralizing dental epithelial organoids (DEOs) from adult dental epithelial stem cells (aDESCs) isolated from mouse incisor tissues. DEOs expressed ameloblast markers, could be maintained for more than five months (11 passages) in vitro in media containing modulators of Wnt, Egf, Bmp, Fgf and Notch signaling pathways, and were amenable to cryostorage. When transplanted underneath murine kidney capsules, organoids produced OHAp crystallites similar in composition, size, and shape to mineralized dental tissues, including some enamel-like elongated crystals. DEOs are thus a powerful in vitro model to study mineralization process by dental epithelium, which can pave the way to understanding amelogenesis and developing regenerative therapy of enamel.

LIMP-2 enhances cancer stem-like cell properties by promoting autophagy-induced GSK3β degradation in head and neck squamous cell carcinoma / 国际口腔科学杂志·英文版

Cancer stem cell-like cells (CSCs) play an integral role in the heterogeneity, metastasis, and treatment resistance of head and neck squamous cell carcinoma (HNSCC) due to their high tumor initiation capacity and plasticity. Here, we identified a candidate gene named LIMP-2 as a novel therapeutic target regulating HNSCC progression and CSC properties. The high expression of LIMP-2 in HNSCC patients suggested a poor prognosis and potential immunotherapy resistance. Functionally, LIMP-2 can facilitate autolysosome formation to promote autophagic flux. LIMP-2 knockdown inhibits autophagic flux and reduces the tumorigenic ability of HNSCC. Further mechanistic studies suggest that enhanced autophagy helps HNSCC maintain stemness and promotes degradation of GSK3β, which in turn facilitates nuclear translocation of β-catenin and transcription of downstream target genes. In conclusion, this study reveals LIMP-2 as a novel prospective therapeutic target for HNSCC and provides evidence for a link between autophagy, CSC, and immunotherapy resistance.

Clinical analysis of twin-to-twin transfusion syndrome combined with necrotizing enterocolitis / 中国小儿急救医学

Objective:To investigate clinical characteristics and high-risk factors of prognosis of twin-to-twin transfusion syndrome(TTTS) combined with necrotizing enterocolitis(NEC).Methods:The clinical data of 102 children with TTTS admitted to the NICU at the Third Affiliated Hospital of Zhengzhou University from January 2017 to January 2020 were collected.Fifty-one pairs(102 cases)of twins without TTTS who were hospitalized at the same time and in the same gestational age were selected as the control group, and the relevant case data were collected and compared.The clinical data of 14 children with NEC in TTTS group were analyzed retrospectively.Results:(1)The average gestational age of TTTS group was(32.24±2.12)weeks, and that was (32.47±1.84) weeks in control group, with no statistical significance( P>0.05). The average birth weight of TTTS group was(1 547.63±523.80)g, which was lower than that of control group(1 658.71±454.13)g( P<0.05). There were 14 children in TTTS group with NEC, with an incidence of 13.7%(14/102), and seven children in the control group with NEC, with an incidence of 6.9%(7/102)( P<0.05). The proportion of very low birth weight infants, NEC occurrence within 2 weeks and mortality in TTTS group were higher than those in control group( P<0.05). (2)Compared with the non-NEC group, the NEC group of TTTS children had lower birth weight, the incidence of intrauterine distress and severe postnatal asphyxia, and the rate of sepsis were significantly higher than those in non-NEC group( P<0.05). (3)Among TTTS children, NEC was diagnosed in ten donors(71.4%) and four recipients(28.6%), with statistically significant difference between two groups( P<0.05). (4)The early clinical symptoms of TTTS complicated with NEC were mainly bloody stools, abdominal distension, poor response, apnea, and vomiting. Conclusion:TTTS is one of the risk factors for NEC, which the occurrence time of TTTS combined with NEC is not completely consistent with the classic NEC, which is more likely to occur within 2 weeks after birth.Children with TTTS complicated with NEC mostly occur in donor infants, and fetal distress in utero, severe asphyxia and sepsis are the high risk factors.The early clinical symptoms of TTTS combined with NEC are not significantly different from those of common NEC, mainly including bloody stools, abdominal diste, poor response, apnea, and vomiting.Vigilance should be raised when similar digestive symptoms appear in children.

Current Opinion in Molecular Pathology of Ameloblastoma: A Literature Review

Ameloblastoma is the most representative epithelial odontogenic tumor in the craniofacial region. Through several studies on Ameloblastoma that have been conducted so far, we have been able to get closer to the reality of Ameloblastoma. However, groundbreaking insight into the pathophysiology of Ameloblastoma has not yet been provided.This review assessed three aspects of five recently published papers on Ameloblastoma: cancer stem cells, calcium signaling, and tumor microenvironment, and compared them with previous studies on tumor physiology, including cancer. In addition, the characteristics of Ameloblastoma revealed by the experimental methods presented in the currently published five papers provide the possibility of Ameloblastoma as a study model in general tumor or cancer studies. Furthermore, the mechanisms of action of the chemicals identified in the studies support their potential as candidates for the second-line treatment of Ameloblastoma.

Clinical analysis of 23 cases of carbapenem-resistant klebsiella pneumoniae sepsis in premature infants / 中国新生儿科杂志

Objective To study the clinical manifestations, risk factors, treatment and prognosis of carbapenem-resistant klebsiella pneumoniae (CRKP) sepsis in premature infants. Method A retrospective analysis was done for the premature infants diagnosed with klebsiella pneumoniae sepsis and admitted to the neonatal wards of the Hospital from April 2015 to March 2018. According to the results of drug sensitive test, the infants was assigned to CRKP group and non-CRKP group. The perinatal factors, clinical manifestations, treatment, and prognosis of the two groups were analyzed. Furthermore, high risk factors for CRKP group were analyzed. Result A total of 39 premature infants with KP sepsis were included in our study. There were 23 cases in the CRKP group and 16 cases in the non-CPAP group. In CPKP group, the gestational age was (29.5 ± 0.6) weeks, the birth weight was (1177 ± 272) g. In non-CRKP group, the gestational age was (30.0 ± 0.5) weeks, the birth weight was (1387 ± 220) g. Univariate Logistic regression analysis showed that low birth weight was a risk factor for CRKP sepsis in premature infants (OR=1.203, 95％CI 1.068～1.355, P=0.002). The proportion of that required combination treatment with antibiotics and the incidence of intracranial hemorrhage after infection in the CPKP group were both higher than that in the non-CRKP group (P<0.05). The proportion and duration of antibiotics used in the first week before the onset of infection in infants with CRKP sepsis and combined antibiotic treatment were significantly higher than those in infants with CPKP sepsis and single antibiotic treatment. The use of antibiotics in the first week before the onset of infection was an independent risk factor for the combined drug treatment of premature infants with CRKP sepsis (OR=10.500, 95％CI 1.015～108.577, P=0.049). In the CRKP group, the improvement rate was 87.0％(20/23), 2 cases were withdrew, and 1 case deceased. In the non-CPKP group, the improvement rate was 87.5％(14/16), and 2 deceased. Conclusion The lower the birth weight, the greater the risk of infection with CRKP sepsis. The proportion of need combination treatment with antibiotics is high in infants with CRKP sepsis. The use of antibiotics in the first week before the onset of infection is a risk factor for combined antibiotic treatment in premature infants with CRKP sepsis .

Analysis of clinical application value combined diagnosis of solitary pulmonary nodules marked tumor / 国际检验医学杂志

Objective To investigate the combined detection of serum cancer antigen(CEA)blank,carbohydrate antigen 125 (CA125)and cytokeratin 19 fragment(CYFRA21-1),neuron specific enolase(NSE)in the diagnosis of solitary pulmonary nodules. Methods 150 cases of patients with solitary pulmonary nodules diagnosed in our hospital from January 2015 to March 2017 were selected as the research subjects,including 51 lung cancer patients(Lung Cancer Group)and 51 benign lung disease patients(benign group).60 healthy people in the same period were selected as control group.The serum levels of CEA,CA125,CYFRA21-1 and NSE were detected in three groups of patients,and the clinical value of the combined detection of various indexes in the differential diagnosis of solitary pulmonary nodules was analyzed.Results The serum levels of CEA,CA125,CYFRA21-1 and NSE in patients with lung cancer were significantly higher than those in the benign and control groups(P<0.05);The sensitivity of CEA,CA125, CYFRA21-1 and NSE in differential diagnosis of malignant solitary pulmonary nodules was 32.32%,27.27%,33.33% and 40.40%,respectively.The specificity was 62.75%,52.94%,60.78%,70.59,respectively;The sensitivity of CEA + NSE+ CY-FRA21-1+CA125 in the differential diagnosis of malignant solitary pulmonary nodules was 81.82%,the specificity was 88.24%, the rate of missed diagnosis was 18.18%,the misdiagnosis rate was 11.76%,the positive predictive value was 93.10%,and the negative predictive value was 71.43%.Conclusion The combined detection of serum CEA,CA125,CYFRA21-1 and NSE has im-portant clinical value in the differential diagnosis of malignant solitary pulmonary nodules.

Effect of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced RSC-364 / 中国药理学通报

Aim To observe the influence of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced rat fibroblast-like synovial cell line RSC-364.Methods The secretion levels of MMP-1, MMP-3, MMP-9 and TIMP-1 were determined using ELISA;MMP-3 and MMP-9 mRNA expressions were detected by RT-PCR.Results MMP-3 and MMP-9 could not be examined in RSC-364 incubated with CCK-8 and unstimulated RSC-364, which was able to product a little MMP-1, TIMP-1 and express even less MMP-3,-9 mRNA.CCK-8 inhibited the increase in MMP-1, MMP-3, MMP-9 secretion and MMP-3,-9 mRNA expression in TNF-α-induced RSC-364.TIMP-1 production was also increased in TNF-α-induced RSC-364.CCK-8 had no effect on TIMP-1 production in TNF-α-induced RSC-364, but was able to reduce the ratios of MMP-1, MMP-3, MMP-9 to TIMP-1.Conclusion The inhibitory effect of CCK-8 on MMPs activity may be related to the decrease of MMPs mRNA expression, MMPs secretion and the ratios of MMPs to TIMP-1 in TNF-α-induced RSC-364, which indicates that CCK-8 might be a possible regulator in the pathogenesis of rheumatoid arthritis.

Mutation analysis of autosomal and X chromosomal STR in gynecologic and breast cancer / 中国法医学杂志

Objective To explore the mutation types and disciplines of STR commonly used in forensic in gynecologic and breast cancerand investigate the application of microdissection in forensic practice involving tumor tissue. Methods DNA of tumor tissues, adjacent normal tissues and peripheral blood from 62 patients with breast cancer, 62 patients with gynecologic cancer and 10 patients with benign gynecologic tumor were amplified by PowerPlex 21 System kit and Argus X-12 kit. Capillary electrophoresis of PCR products was carried out on an ABI 3130 Genetic Analyzer to obtain genotypes. Some tumor tissues with STR variation were microdissected. Results The genotype of peripheral blood in cancer patient was consistent with that of corresponding normal tissue. 4 types of STR variations were found in 46.77% gynecologic cancer tissues, compared with that in benign tumor tissues and breast cancer, the difference of STR variation was significant(P<0.01,P=0.009). The genotype of stromal cells separated by microdissection was consistent with that of corresponding adjacent normal tissue. Conclusion The STR loci detected in the study with poor stability are not suitable for forensic cases involving gynecologic cancer tissues. The genotype of stromal cells separated accurately from tumor tissues by microdissection could represent the normal DNA genotype of the individual with cancer. Microdissection is an effective solution in forensic cases with tumor tissues.

Comparison of detection of SNP and STR in formalin-fixed tissue / 中国法医学杂志

Objective The purpose of this study was to detect the degradation degree of long-term formalin fixed tissue and to compare the detection rate of STR with SNP. Methods DNA was extracted from 24 formalin-fixed tissues stored at -20 ℃ for five years, and the concentration and degradation index of DNA was quantified with Quantifiler? Trio DNA Kit. A 55-SNP multiplex SNaPshot assay and PowerPlex? 21 system were used to amplify SNP and STR loci, respectively. Results The results showed that the degradation indexes of 24 specimens were ranged from 1~8. The SNP genotypes of the 24 specimens were completely consistent with the non-degraded DNA from the same individuals and the successful genotyping rate was 100%. However, 33 allele dropouts were observed with STR genotyping in 8 samples, of which the degradation index was higher than 2.6, and the fragment size of the 75.8% allele was longer than 300bp. The likelihood ratio based on 16 typable STR loci in the sample was close to that onthe basis of 54 SNPs. However, likelihood ratio based on more than 17 STR loci was over that accord to 54 SNPs. There was a negative correlation between the fragment size of STR and the allele detection rate, and a negative correlation also observed between the degradation index of samples and the allele detection rate except for two samples with mild degradation. Conclusion This study validated that the long-term formalin-fixed tissues were susceptible to degradation, and the SNP was more suitable for detecting these tissues than STR typing system. However more SNP loci are needed to test in order to increase the discrimination power.

Levels of 25-(OH)D in 0 ～ 12 years Old Children in Kunming / 昆明医科大学学报

Objective To learn the levels of serum 25-(OH)D in 0 12 years old chindren in Kunming.Methods Serum 25-(OH)D levels of children (4 498) were measured by using Roche cobas 8 000 / E602 automatic electrochemical luminescence immunoassay.Results As the increase of age,the levels of serum 25-(OH)D were gradually decreased,different age groups had significant difference (P＜0.05).The levels of 25-(OH)D between boys and girls had no significant difference (P＞0.05);The average level of sernm 25-(OH)D was 83.48 nmol/L,the level of serum 25-(OH)D was distributed in 0 ～ 1 years old group,and most concentrated in the 6 ～ 9 years old group.Conclusion The levels of 25-(OH)D in 0 ～ 12 years old chindren are in a good status,but with the increase of age,the level of 25-(OH)D is decreasing gradually,especially the levels of 25-(OH)D decreased significantly after the age of three,in shortage or lack of status,this should cause everybody's attention.

Study of genetic polymorphisms of 12 short tandem repeats on X chromosome in ethnic Han population from Hebei Province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate genetic polymorphisms of 12 X chromosome short tandem repeat (X-STR) loci in ethnic Hebei Han population using an Investigator Argus X-12 amplification kit.</p><p><b>METHODS</b>DNA was extracted for 198 unrelated individuals (96 males and 102 females) and amplified with a fluorescence labeled multiplex PCR system. PCR products were separated and genotyped with capillary array electrophoresis.</p><p><b>RESULTS</b>Only DXS10103 and DXS10101 showed significant linkage disequilibrium at the 12 X-STR loci. One hundred and forty-eight alleles, including 22 off-ladder (OL) alleles, were observed at the 12 X-STR loci in the population. The heterozygosity and polymorphic information content (PIC) were 0.5074-0.9143 and 0.4377-0.9079, respectively. The power of discrimination (PD) was 0.5074-0.9143 in males and 0.6876-0.9863 in females. The mean exclusion chance was 0.4377-0.9079 in the trios cases and 0.2984-0.8373 in the duo cases, respectively.</p><p><b>CONCLUSION</b>The Investigator Argus X12 amplification system is highly polymorphic in ethnic Han population from Hebei and is useful for personal identification and paternity testing.</p>

Effects of CCK-8 and its receptor antagonists on expression of CREB and pCREB in prefrontal cortex and hippocampus of morphine withdrawal rats / 中国病理生理杂志

AIM:To observe the effects of cholecystokinin octapeptide (CCK-8) and its receptor antagonists on cAMP response element binding protein ( CREB) and phosphorylated CREB ( pCREB) expression in frontal cortex and hippocampus of morphine withdrawal rats , which aim to explore the post-receptor mechanism through which CCK-8 regu-lates morphine withdrawal .METHODS: After the morphine dependence and naloxone-precipitated withdrawal animal models were established, the effects of CCK-8, L-364718 (CCK1 receptor antagonist) and LY-288513 (CCK2 receptor an-tagonist) pretreatment on CREB and pCREB expression in frontal cortex and hippocampus were observed by Western blot -ting and immunohistochemistry .RESULTS:In rat frontal cortex neuron , CREB was expressed in both cytoplasm and nu-cleus, but pCREB was only highly expressed in the nucleus .In the pyramidal cell layer of hippocampal CA 1 region, CREB showed high expression in the cytoplasm and low expression in the nucleus , while pCREB was only expressed in the nu-cleus.No obvious change of CREB was observed after either chronic morphine treatment or naloxone withdrawal .The pCREB expression was increased after chronic morphine treatment and further increased after naloxone withdrawal .Com-pared with the withdrawal group , chronic pretreatment with CCK-8, L-364718 and LY-288513 had no effect on CREB expression in the frontal cortex , but obviously decreased the pCREB expression .In the hippocampus , pretreatment with L-364718 and LY-288513 decreased CREB and pCREB expression , but only the pCREB expression was decreased after CCK-8 treatment.CONCLUSION:CCK-8 and CCK receptor antagonists may alleviate morphine withdrawal symptoms by regulating CREB , with specificity in different brain regions .

Critical Amino Acids Analysis of Epitopes in Pen a1 / 分析化学

AscreeningmethodforcriticalaminoacidsinEpitopesusinganti-Epitopeantibodywasdeveloped. The amino acids' frequency of occurrence in Epitopes of shrimp antigen Pen a1 was calculated using MEGA5 software and their conservative property of tropomyosin in SDAP database bank was analyzed using DNAMAN software. Potential critical amino acids based on these two methods were identified, namely Epitope 1 ( K, E, N) , Epitope 2 ( K, L, E) , Epitope 3 ( E, R, D, L, Q) , Epitope 5 ( K, L, Q) . The mutated polypeptides corresponding to these Epitopes in which these critical amino acids were substituted with alanine were synthesized. The IgG binding ability of these mutants was analyzed by competitive immunodot-blot method and indirect ELISA using Epitope antibody to screen the critical amino acids. Experimental results showed that the critical amino acids of the four Epitopes were as follows: Epitope 1 ( Gln ) , Epitope 2 ( Leu and Glu ) , Epitope 3 ( Leu and Asp) , Epitope 5 ( Leu) . The feasibility of this screening method was proved and it also offered a theoretical foundation for research on sensitization mechanism of Pen a1 and desensitization using gene modification.

Keyhole limpet hemocyanin induced Th1/Th2 imbalance in splenocytes of Balb/C mice / 中国组织工程研究

BACKGROUND:Establishing a characteristic.stable and repeatable model of Th1/Th2 imbalance in animals,is the key of studying the mechanism of Th1/Th2 imbalance.OBJECTIVE:To observe the characteristics of Th1/Th2 imbalance in splenocytes derived from Balb/C mice immnnized by keyhole limpet hemocyanin(KLH).DESIGN:A randomized control exploratory experiment.SETTING:Hebei Provincial Forensic Laboratory.Institute of Basic Medicine,Hebei Medical University.MATERLALS:The experiment was carried out in the Hebei Provincial Forensic Laboratory,Institute of Basic Medicine,Hebei Medical University from September 2005 to January 2007.Balb/C mice were adopted in this study.and all the disposals were in accordance with the guidance of animal ethics.METHODS:Balb/C mice were immunized with KLH emulsified in complete Freund's adjuvant(CFA),splenocytes were acquired,and the peak of cytokine secretion was determined in 3 groups:KLH groups of 6.25 mg,kg.12.5 mg,kg and 25 mg/kg.According to the immunizing dose and immunizing frequency.mice were divided into 7 groups:KLH groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,secondary immunity groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,as well as control group.According to the determined levels of IgG1 and IgG2a in blood serum.mice were divided into KLH group of 6.25 mg/kg and control group.MAIN OUTCOME MEASURES:Mice splenocytes supematant was detected with enzyme linked immunosorbent assay (ELISA)for the production of Th1 cytokines interferon (IFN)-γ,interleukin(IL)-2.IL-12 p40 and Th2 cytokines IL-4 and IL-5.The levels of Th1 antibody IgG2a and Th2 antibody IgG1 in blood serum were also detected by ELISA.RESULTS:The spleen derived from KLH-immunized mice appeared hypertrophy,and the number of splenocytes was manifold.Splenocytes restimulated with KLH in vitro produced much more IFN-γ,IL-2,IL-4,IL-5,but not IL-12p40.IL-2 secretion was obviously elevated after incubated for 24 hours and achieved pinnacle at 48 hours;productions of IL-4,IL-5 and IFN-γ were elevated after 24 hours,and increased gradually to 96 hours;IL-12p40 production was very low at every time point.Using different doses of KLH inlmunity once or twice,could all lead to the elevated productions of IL-2,IL-4,IL-5,IFN-γ,and the elevation of IL-4/IFN-γ ratio,but the secondary immunity group of 6.25 mg/kg KLH showed obviously higher levels than other groups(P＜0.01).The level of KLH specific antibody IgG2a and IgG1,especially IgG1 was elevated in blood serum of KLH-immunized mice.CONCLUSION:Balb/C mice immunizad with KLH emulsified in CFA can indce a Th2 predominant imbalance in splenocytes.

Effects of cholecystokinin octapeptide on TNF- α- induced IL- 6 expression and its possible molecular mechanismin rat synovial cell strain RSC-364 / 中国病理生理杂志

AIM: To investigate the effect of sulfated cholecystokinin octapeptide (CCK -8 ) on TNF -α induced IL - 6 mRNA expression, NF - κB activation in the rat fibroblast - like synovial cell strain RSC - 364 and its possible receptor mechanisms. METHODS: RSC -364 cells were stimulated with TNF - α( 10 μg/L) in the presence or absence of sCCK- 8( 10-8 - 10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L). IL -6 and CCK receptor A/B (CCK- AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction (RT- PCR) at 3 h after stimulation, and nuclear factor - κB (NF - κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at lh after stimulation. At 30 min of stimulation the IκB protein level in cytoplasma was measured by Western blotting. RESULTS: Both CCK - AR and CCK - BR were constitutively expressed on RSC - 364. sCCK - 8, at concentrations from 10-8 mol/L to 10 -6 mol/L, significantly increased IL - 6 mRNA expression, CCK - AR and CCK - BR mRNA expression, NF - κB binding activity and IκB protein degradation. The effects of sCCK - 8 on NF - κB activity and IκB degradation level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: sCCK - 8 upregulats TNF - α- induced IL - 6 mRNA expression by NF - κB pathway through its receptor on rat synoviocytes, suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.

Diallyl trisulfide inhibites interleukin-1? expression induced by lipopolysaccharide through inhibiting NF-?B activation in murine lung / 基础医学与临床

Objective To investigate the role of nuclear factor-kappa B(NF-?B)in the modulation of diallyl trisulfide(DATS)on interleukin-1?(IL-1?)expression induced by lipopolysaccharide(LPS)in mice with acute lung injury(ALI).Methods Mice were randomly divided into Control group,ALI group,DATS group,DATS prevention group and DATS treatment group.The expression of IL-1? mRNA in the lung tissue was detected by reverse transcription PCR(RT-PCR).NF-?B activity in the lung tissue was detected by electrophoresis mobility shift assay(EMSA).The expression of phospho-I?B and I?B were assayed by Western blot.Results The expression of IL-1? mRNA,NF-?B activity and the phospho-I?B expression in lung tissues increased significantly at ALI group(P

Effects of DAG-PKC signaling pathway on cAMP-PKA signaling pathway in anti-inflammatory effects of CCK-8 / 中国药理学通报

0.05).But PMA increased and SC-3088 decreased cAMP content and PKA activity in LPS-stimulated rat PIMs(P

Effect of CCK-8 on IL-1?-induced RSC-364 proliferation / 中国药理学通报

Aim To investigate the effects and mechanisms of CCK-8 on IL-1? induced proliferation of RSC-364, a rat fibroblast-like synovial cell line. Methods MTT colorimetric assay and Western blot were used to measure cell proliferation and p38MAPK phosphorylation level to elucidate the mechanism of CCK-8 in IL-1? induced RSC-364 proliferation. Results CCK-8 significantly inhibited IL-1?-induced RSC-364 proliferation at 10 -12 , 10 -10 , 10 -8 , 10 -6 mol ? L -1 , and IL-1?-activated p38MAPK activity at 10 -10 , 10 -8 , 10 -6 mol?L -1 in a dose-dependent manner. The effect of CCK-8 was blocked by CR1409 (a CCKA-receptor antagonist) and CR2945 (a CCKB-receptor antagonist). Conclusion CCK-8 inhibits IL-1?-induced RSC-364 proliferation, probably by reducing p38MAPK activity through CCKA and CCKB receptors.

Application of case-based PBL teaching method in forensic toxicology / 中华医学教育探索杂志

Forensic toxicology is a specialized subject which forensic medical appraisers and students must learn and master. As a new teaching mode,problem based learning method is attrcting a lot of attention. Applying "case" based PBL teaching method is an important method to train innovative and high technical ability forensic talents.

Cholecystokinin octapeptide inhibits the in vitro expression of CD14 in rat pulmonary interstitial macrophages induced by lipopolysaccharide / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of cholecystokinin octapeptide (CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophages (PIM) in vitro.</p><p><b>METHODS</b>PIM were isolated and cultured in the presence or absence of LPS, CCK-8, proglumide (the antagonist of CCK receptors) and vehicle. The expression of membrane CD14 (mCD14) protein was assayed by flow cytometry and soluble CD14 (sCD14) in the supernatant was analyzed semi-quantitatively by Western blot. TNF-alpha in the supernatant was detected with ELISA.</p><p><b>RESULTS</b>CCK-8, at concentrations of 10(-7) mol/L and 10(-6) mol/L, significantly inhibited the expression of mCD14. Release of sCD14 and TNF-alpha in the supernatant was up-regulated by LPS (1 microg/ml) but reduced by CCK-8. The effect of CCK-8 was inhibited by proglumide.</p><p><b>CONCLUSION</b>CCK-8 negatively modulated several functions of LPS-stimulated PIM through CCK receptors. This may be one of the mechanisms for CCK-8 to alleviate inflammation in lung tissue during endotoxemia.</p>