Investigation of patients’preferences regarding therapeutic drug monitoring of mycophenolic acid in solid organ transplantation / 中国药房

OBJECTIVE To investigate the preferences of patients who underwent solid organ transplantation regarding therapeutic drug monitoring （TDM） of mycophenolic acid （MPA） and explore the factors influencing patients’ decision-making process， so as to provide support for the development of individualized medication guidelines for MPA and improvement of clinical decision-making. METHODS The cross-sectional study was used to design the questionnaire on the patients’ preferences to accept MPA TDM， and involved patients who underwent solid organ transplantation and received MPA treatment at two tertiary hospitals in Beijing from April 14， 2022， to June 27， 2022. The Likert 5-level scoring method was used to score the patients’ preferences to accept MPA TDM， the influencing factors and their correlation of the patients’ preferences to accept MPA TDM were analyzed by Pearson correlation analysis and binary Logistic regression analysis， and the nonparametric test and chi-square test were used to rank and analyze the consistency of the factors affecting patients’ preference decision. RESULTS A total of 140 questionnaires were collected， and the effective recovery rate was 77.35%. The average preference score of 140 patients to receive MPA TDM was （4.01±0.65）， and the overall preference value was high. There were 116 （82.86%） patients agreed or strongly agreed with MPA TDM. Significant differences were observed in preference scores between patients who had previously undergone MPA TDM and those who had never undergone it （[ 4.30±0.53） scores vs. （3.80±0.65） scores， P＜0.001]. Additionally， patients’ preference scores were significantly influenced by their understanding level and attention level （P＜0.001）. The ranking of factors contributing to decision-making exhibited consistency （P＜0.001）. The factors were ranked in descending order of clinical efficacy， safety， comfortability， economy and time cost. CONCLUSIONS The patients who underwent solid organ transplantation hold high preferences towards MPA TDM. The primary factors influencing their decisions are their prior experience， understanding level， and attention level.

Preliminary research on breast cancer organoids as a radiobiological model / 中华放射肿瘤学杂志

Objective:To construct breast cancer organoid culture system and conduct histological characterization and preliminary radiobiological characteristics study.Methods:Different molecular types of breast cancer cell lines and patient-derived tumor cells were cultured in vitro to form breast cancer organoids and characterize their tissue structure. In addition, Ki-67, ER, PR and Her2 markers were evaluated by immunohistochemical staining. Breast cancer organoids were irradiated with 4 Gy and 8 Gy. The numbe and diameter changes of breast cancer organoids at 0, 24, 48 and 96 h after irradiation were observed to evaluate the irradiation-induced damage to the organoids. Results:Breast cancer cell lines and patient-derived tissues formed organoid structures at 6 d. HE staining showed the microstructures, and the expression profile of markers was spatially heterogeneous. The expression patterns of markers were similar between patient-derived organoids and original tumor tissues. Irradiation of MCF-7 breast cancer organoids led to growth arrested, and some of the formed organoids collapsed and the proliferating trend gradually recovered from 48 h to 96 h. MDA-MB-231 breast tumor organoids showed radioresistance, growth arrested, but the structures remained intact, the recovery trend was still not observed at 96 h. The tissue-derived organoids from triple-negative patients also showed radiation tolerance. After irradiation, the organoids continued to grow without significant structural changes, whereas the growth trend was significantly smaller than that in the non-irradiated group.Conclusions:Breast cancer organoids formed by in vitro culture of breast cancer cells from different sources and different molecular types have microstructure and heterogeneity, which can reflect the expression of source tissue markers and show different radioresistance. Organoids derived from triple-negative breast cancer are more resistant to irradiation.

Advances in the diagnosis and treatment of occult inguinal hernia / 国际外科学杂志

With the advent of laparoscopic inguinal hernia repair, occult inguinal hernias not reported by physical examination are frequently found during surgery. Occult inguinal hernia is an early inguinal hernia, which is easy to be ignored because its clinical symptoms are not obvious. Currently, the best diagnostic method for occult inguinal hernia is laparoscopy, Noninvasive tests such as ultrasound and MRI should be performed in patients with first symptoms. With the increase of age, abdominal wall strength decrease, and the proportion of occult inguinal hernia requiring treatment will also increase, doctors need to choose a reasonable treatment plan according to the actual situation of patients. The key to the diagnosis and treatment of occult inguinal hernia lies in making clear its definition and establishing uniform diagnostic criteria. Through reasonable and accurate preoperative examination combined with laparoscopic techniques and the adjustment of surgical approaches, occultic defects can be found and repaired. Individualized treatment for different populations can avoid future symptoms and reduce additional surgical risks.

Analysis of the effect of face-to-face embracing posture in the washing of external auditory canal in children / 中国实用护理杂志

Objective@#To explore a method to improve the flushing degree of the external auditory canal in children (≤6 years old).@*Methods@#A total of 160 children with external auditory canal irrigation were divided into observation group and control group by stratified randomization grouping method and there was 80 cases in each group. The children in the observation group were treated with family and children in a face-to-face sitting posture for external auditory canal irrigation. The children in the control group were in a traditional sitting position, and the children were placed on the family′s legs for external ear canal irrigation. The two groups were combined with psychological care and then compared the two groups treatment effect and the satisfaction rate.@*Results@#In the observation group, 81.25%(65/80) children completed the external auditory canal irrigation treatment in one time. In the control group, 48.75%(39/80) children completed the external auditory canal irrigation treatment in one time (χ2 value was 11.218, P<0.01). The compliance rate of one-time external auditory canal irrigation was 97.5%(78/80) in the observation group and was 83.75%(67/80) in the control group, there was significant difference between the two groups (χ2 value was 8.901, P<0.01). The satisfaction rate of children`s families was 98.75%(79/80) in the observation group and 90.00%(72/80) in the control group, there was significant difference between the two groups (χ2 value was 5.769, P<0.05).@*Conclusions@#The use of face-to-face sitting posture for external auditory canal irrigation has the advantages of high fitness and good safety and improves the efficiency of flushing treatment.

Analysis of the effect of face-to-face embracing posture in the washing of external auditory canal in children / 中国实用护理杂志

Objective:To explore a method to improve the flushing degree of the external auditory canal in children (≤6 years old).Methods:A total of 160 children with external auditory canal irrigation were divided into observation group and control group by stratified randomization grouping method and there was 80 cases in each group. The children in the observation group were treated with family and children in a face-to-face sitting posture for external auditory canal irrigation. The children in the control group were in a traditional sitting position, and the children were placed on the family′s legs for external ear canal irrigation. The two groups were combined with psychological care and then compared the two groups treatment effect and the satisfaction rate.Results:In the observation group, 81.25%(65/80) children completed the external auditory canal irrigation treatment in one time. In the control group, 48.75%(39/80) children completed the external auditory canal irrigation treatment in one time ( χ2 value was 11.218, P<0.01). The compliance rate of one-time external auditory canal irrigation was 97.5%(78/80) in the observation group and was 83.75%(67/80) in the control group, there was significant difference between the two groups ( χ2 value was 8.901, P<0.01). The satisfaction rate of children`s families was 98.75%(79/80) in the observation group and 90.00%(72/80) in the control group, there was significant difference between the two groups ( χ2 value was 5.769, P<0.05). Conclusions:The use of face-to-face sitting posture for external auditory canal irrigation has the advantages of high fitness and good safety and improves the efficiency of flushing treatment.

Survey on the quadrivalent influenza vaccine intention and related factors of health care workers in the Pearl River Delta region from 2015 to 2017 / 中华预防医学杂志

Objective@#To analyze the quadrivalent influenza vaccine intention of 718 health care workers (HCWs) in the Pearl River Delta region from 2015 to 2017.@*Method@#In May 2018, 718 HCWs from the department related to the diagnosis and treatment of influenza in 17 hospitals (6 tertiary hospitals, 5 secondary hospitals and 6 primary hospitals) from Guangzhou, Jiangmen, Zhuhai and Dongguan were selected by using stratified sampling method. Questionnaire survey and face-to-face interview were used to collect the information of influenza vaccination, the intention of the quadrivalent influenza vaccine, the acceptance of free and required vaccination policies, and recommendations for increasing influenza vaccination intentions from 2015 to 2017. The multivariate logistic regression was used to analyze factors associated with the vaccination intention.@*Results@#A total of 718 HCWs were surveyed and 147 of them were interviewed face to face. Among them, the vaccination rate of primary hospitals [17.39%(40/230)] was higher than that of other hospitals (χ2=15.80, P<0.05). If the vaccine could be free, 84.82% (609/718) of HCWs would like to be vaccinated. The multivariate logistic regression showed that the factors, HCWs who were aged ≥50 years (OR=3.44, 95%CI:1.43-8.28), worked in department of prevention and health care (OR=2.35, 95%CI:1.16-4.75), learned about the quadrivalent influenza vaccine (OR=2.94, 95%CI:2.08-4.18), knowed that HCWs are priority (OR=2.33, 95%CI:1.56-3.48), and had a history of trivalent influenza vaccination from 2015 to 2017 (OR=4.70, 95%CI:3.08-7.15), were associated with the vaccination intention.@*Conclusion@#HCWs in the Pearl River Delta region had weak inclination of getting quadrivalent influenza vaccine. HCWs who were age (≥50 years old), worked in department of prevention and health care, learned about the quadrivalent influenza vaccine, knowed that HCWs are priority, and had a history of trivalent influenza vaccination from 2015 to 2017 were factors positively associated with the vaccination intention.

Advances in research on treatment of aberrant crypt foci / 实用肿瘤学杂志

Aberrant crypt foci(ACF)is highly similar to colorectal cancer(CRC)in phenotypic and molecular changes,and is positively correlated with the risk of CRC. It is considered as precancerous lesion of CRC. Many compounds have therapeutic effects on ACF,inhibiting their occurrence and malignant transformation,which can be regarded as a preventive effect on CRC. In this paper,we will review the treatment of ACF by various compounds in different ways in recent years.

Effects of tumor-associated fibroblasts in the development of colorectal cancer and its research progress / 实用肿瘤学杂志

Cancer-associated fibroblasts(CAFs)are one of the most important cellular components in the external microen-vironment of tumors. CAFs can interact with tumor cells and secrete a variety of soluble factors such as growth factors,chemokines,and so on. CAFs are also involved in the proliferation,invasion,migration,metastasis and drug resistance of colorectal cancer through regu-lating multiple signaling pathways that play a key role in the progression of colorectal cancer. Recent studies have found that CAFs re-lated markers and genes can be used as reference indicators for the predicting prognosis of colorectal cancer. Therefore,targeting CAFs may be a key target for early diagnosis,treatment and prognosis of colorectal cancer. This article summarizes the characteristics of CAFs,recruitment and activation process,and its role in the development of colorectal cancer in order to provide a new scientific di-rection for the mechanism of action and clinical application of CAFs in colorectal cancer.

Mechanism for synergistic effect of IRF4 and MITF on tyrosinase promoter / 中南大学学报(医学版)

Objective:To investigate the mechanism for the synergistic effect of interferon regulatory factor 4 (IRF4) and microphthalmia-associated transcription factor (MITF) on tyrosinase (TYR)promoter.Methods:The synergistic transcriptional effect,subcellular localization,and protein-protein interaction for IRF4 and MITF were observed by luciferase assay,immunofluorescence,GST-pull down,and co-immunoprecipitation,respectively.Results:IRF4 and MITF proteins were co-expressed in the cell nucleus.IRF4 augmented the transcriptional function of MITF (but not the mutant MITF) to activate the expression of the TYR promoter,but with no effect on other MITF-specific target promoters.IRF4 alone did not affect TYR promoter significantly.No direct interaction between the two proteins was noted.Conclusion:IRF4 and MITF exert a specifically synergistic effect on activation of TYR promoter through IRF4-mediated upregulation of transcriptional function of MITF.This synergistic effect is mainly regulated by MITF;DNA might be involved in the interaction between the two proteins.

Study of gene mutation and pathogenetic mechanism for a family with Waardenburg syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the pathogenetic mechanism of a family affected with Waardenburg syndrome.</p><p><b>METHODS</b>Clinical data of the family was collected. Potential mutation of the MITF, SOX10 and SNAI2 genes were screened. Plasmids for wild type (WT) and mutant MITF proteins were constructed to determine their exogenous expression and subcellular distribution by Western blotting and immunofluorescence assay, respectively.</p><p><b>RESULTS</b>A heterozygous c.763C>T (p.R255X) mutation was detected in exon 8 of the MITF gene in the proband and all other patients from the family. No pathological mutation of the SOX10 and SNAI2 genes was detected. The DNA sequences of plasmids of MITFand mutant MITFwere confirmed. Both proteins were detected with the expected size. WT MITF protein only localized in the nucleus, whereas R255X protein showed aberrant localization in the nucleus as well as the cytoplasm.</p><p><b>CONCLUSION</b>The c.763C>T mutation of the MITF gene probably underlies the disease in this family. The mutation can affect the subcellular distribution of MITF proteins in vitro, which may shed light on the molecular mechanism of Waardenburg syndrome caused by mutations of the MITF gene.</p>

Clone and prokaryotic expression of staphylococcus aureus drug resistance adenylyltransferase gene / 国际检验医学杂志

Objective To clone and express Staphylococcus aureus drug resistance adenylyltransferase gene in E .coli BL21 ,and to make the foundation for its function research .Methods Primers were designed on the basis of adenylyltransferase gene in gen‐bank ,PCR was used to amplify adenylyltransferase gene using Staphylococcus aureus genomic DNA as template .The obtained PCR production was attatched with pGEX‐4t‐1(+ ) plasmid ,and transformed into E .coli BL21 (DE3) .The recombinant plasmid was di‐gested by double enzyme digestion and identified by gene sequence .The recombinant protein was induced to expression by IPTG and identified by Western‐blotting .Results Using Staphylococcus aureus genome as a template ,the target fragment about 800 bp was successful amplified .After enzyme‐cutting and DNA‐sequencing ,the target fragment showed that the ORF begin with ATG ,end with TAG ,783 bp in length ,the predicted isoelectric point and molecular weight were 7 .75 and 29 × 103 ,and it was homology 99%homology with the reported sequence gene in genbank .SDS‐PAGE and Western‐blot showed the molecular weight of recombinant fusion protein was about 55 × 103 .Conclusion Adenylyltransferase gene of Staphylococcus aureus was successfully cloned and ex‐pressed in E .coli as a fusion protein ,which makes the foundation for the research of its function .

Molecular pathogenesis of Waardenburg syndrome type II resulting from SOX10 gene mutation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the molecular mechanism of Waardenburg syndrome type II (WS2) resulting from SOX10 gene mutation E248fs through in vitro experiment.</p><p><b>METHODS</b>293T cells were transiently transfected with wild type (WT) SOX10 and mutant type (MT) E248fs plasmids. The regulatory effect of WT/MT SOX10 on the transcriptional activity of MITF gene and influence of E248fs on WT SOX10 function were determined with a luciferase activity assay. The DNA binding capacity of the WT/MT SOX10 with the promoter of the MITF gene was determined with a biotinylated double-stranded oligonucleotide probe containing the SOX10 binding sequence cattgtc to precipitate MITF and E248fs, respectively. The stability of SOX10 and E248fs were also analyzed.</p><p><b>RESULTS</b>As a loss-of-function mutation, the E248fs mutant failed to transactivate the MITF promoter as compared with the WT SOX10 (P<0.01), which also showed a dominant-negative effect on WT SOX10. The WT SOX10 and E248fs mutant were also able to bind specifically to the cattgtc motif in the MITF promoter, whereas E248fs had degraded faster than WT SOX10.</p><p><b>CONCLUSION</b>Despite the fact that the E248fs has a dominant-negative effect on SOX10, its reduced stability may down-regulate the transcription of MITF and decrease the synthesis of melanin, which may result in haploinsufficiency of SOX10 protein and cause the milder WS2 phenotype.</p>

The analysis of nystagmus in patients with posterior canal benign paroxysmal positional vertigoin positioning test / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze and summarize nystagmus of patients with posterior canal benign paroxysmal positional vertigo (BPPV) in positioning test,and to improve the diagnosis and treatment of posterior canal BPPV (PSC-BPPV).@*METHOD@#The present study was conducted on 175 patients who had unilateral BPPV of the posterior semicircular canal (PSC). Their positional nystagmus recorded by videnonystagmography in Dix-Hallpike test,roll test and roll over test were analyzed to summarize the characteristics of nystagmus on nystagmograph of PSC-BP-PV.@*RESULT@#Of the 175 patients, lesion was located in the left PSC in 69 (39.4%) patients,the right PSC in 106 (60. 6%)patients. The nystagmus of patients with PSC-canalithiasis showed upward on the vertical phase of nystagmograph and orientated the different side on horizontal phase in the head hangging position. The horizontal phase pointed to the contralateral side in 47(26. 9%) patients, the ipsilateral contralateral side in 100(57. 1%) patients,no significant reverse ingredients in 28(16.0%) patients. When these patients returned to sit,139(79.4%) patients showed down beating positioning nystagmus, whereas 36 (20. 6%) patients with no nystagmus only had a short vertigo or dizziness. The horizontal phase of the 139 patients pointed to the contralateral side in 40(22. 9%) patients,the ipsilateral contralateral side in 68(38. 9%) patients,no significant reverse ingredients in 31(17. 7%) patients. In roll test,12 patients of the right PSC-BPPV presented an up-beating rotatory nystagmus when the head turned to right,and 5 patients of the left PSC-BPPV presented a down-beating rotatory nystagmus when the head turned to left. When the patients changed body from the left lateral position to the right lateral position in the roll over test, 74(42. 3%) patientsshowed vertical positioning nystagmus. In 30 patients who presented an up-beating nystagmus, there were 25(83. 3%) patientscame from the right PSC-BPPV. In 44 patients who presented a down-beating nystagmus, there were 36(81. 8%) patientscame from the left PSC-BPPV. The direction of the vertical nystagmus was highly correlated with the judgment about the side of the PSC-BPPV in roll over test (P<0. 01).@*CONCLUSION@#The patient with PSC-canalithiasis showed an uncertain direction in torsional nystagmus in Dix-Hallpike test,the diagnosis was mainly concern with the vertical nystagmus. When we found a rotatory nystagmus with much more up-beating nystagmus in roll test, it might be PSC-BPPV. We also can use the roll over test to diagnose the location of the otolith in which side of the PSC-BPPV.

The analysis of masking therapy in the early stage of the patients with noise-induced tinnitus / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the effect of masking therapy for the early stage of the patients with noise-induced tinnitus,and imply the treatment for patients with noise-induced tinnitus.@*METHOD@#Sixty-eight cases with tinnitus were studied. All the patients took the audiological examinations and tinnitus tests firstly, and accepted the masking therapy for 6 months. The therapeutic effiency was evaluated according to tinnitus handicap inventory (THI) and subjective visual-analogue scale (VAS). The minimum masking intensity was also evaluated.@*RESULT@#The majority of the patients with noise-induced tinnitus (59 cases, 86. 8%) had tinnitus frequency of 4 kHz,and most of them (44 cases, 64. 7%) had positive residual inhibition tests. Tinnitus completely disappeared in 3 cases after masking therapy, and the efficiency of this treatment is 83. 8%. There was significant difference in the scores of THI and VAS before and after therapy(P<0. 01), and there was also significant difference in the minimum masking intensity (P<0. 01).@*CONCLUSION@#Masking therapy is the most important treatment for the patients in the early stage of noise-induced tinnitus. The therapeutic effiency is significant and should be promoted.

Analysis of nuclear localization and signal function of MITF protein predisposing to Warrdenburg syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the role of dysfunction of nuclear localization signals (NLS) of MITF protein in the pathogenesis of Waardenburg syndrome.</p><p><b>METHODS</b>Eukaryotic expression plasmid pCMV-MITF-Flag was used as a template to generate mutant plasmid pCMV-MITF△NLS-Flag by molecular cloning technique in order to design the mutagenic primers. The UACC903 cells were transfected transiently with MITF and MITF△NLS plasmids, and the luciferase activity assays were performed to determine their impact on the transcriptional activities of target gene tyrosinase (TYR). The oligonucleotide 5'-GAACGAAGAAGAAGATTT-3' was subcloned into pEGFP-N1 to generate recombinant eukaryotic expression plasmid pEGFP-N1-MITF-NLS. The NIH3T3 cells were transfected separately with MITF, MITF△NLS, pEGFP-N1 and pEGFP-N1-NLS plasmids, and their subcellular distribution was observed by immunoflorescence assays.</p><p><b>RESULTS</b>Expression plasmids for the mutant MITF△NLS with loss of core NLS sequence and pEGFP-N1-NLS coupled with MITF△NLS were successfully generated. Compared with the wild-type MITF, MITF△NLS was not able to transactivate the transcriptional activities of promoter TYR and did not affect the normal function of MITF. MITF△NLS was only localized in the cytoplasm and pEGFP-N1 was found in both the cytoplasm and nucleus, whereas pEGFP-N1-NLS was mainly located in the nucleus.</p><p><b>CONCLUSION</b>This study has confirmed the localization function of NLS sequence 213ERRRRF218 within the MITF protein. Mutant MITF with loss of NLS has failed to transactivate the transcriptional activities of target gene TYR, which can result in melanocyte defects and cause WS.</p>

An Analysis of Auditory Brainstem Response in Autistic Children of 6 Years or Younger / 听力学及言语疾病杂志

Objective To investigate the characteristics of auditory brainstem response (ABR) in autistic children of 6 years or younger .Methods A total of 98 children with normal hearing and 22 autistic children were en‐rolled in this study .They were divided into 3 groups :1~2 years age group ,20 .05) .In 80 dB nHL click sound stimulus ,there was also no statistical difference in the values of peak latencies or interpeak intervals of ABR between normal hearing children and autistic children (P> 0 .05) ,whereas ,prolongation of peak latencies or interpeak intervals of ABR occurred in 36 .36%(25 .00% ears) autistic children .The ratio of prolongation of peak latencies of I ,III ,V waves and interpeak inter‐vals of I-III ,III-V ,I-V occurred in 4 .54% ,6 .82% ,13 .64% ,18 .18% ,6 .83% and 15 .91% ,respectively . Conclusion The ABR in some autistic children present abnormality and occur in various forms with the prolongation of interpeak intervals of I -III as the most common .

Construction and analysis of recombinant eukaryotic expression plasmids for SOX10, the causative gene of Warrdenburg syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the exogenous expression and subcellular localization of wild type (WT) and mutant SOX10 proteins in vitro through generation of expression plasmids in order to reveal the pathogenesis of Waardenburg syndrome (WS).</p><p><b>METHODS</b>The plasmids pECE-SOX10 and pCMV-Flag were ligated after they were subjected to double enzyme digestion using molecular cloning technique to generate recombinant eukaryotic expression plasmid pCMV-SOX10-Flag, which was as a template to generate expression plasmids for novel mutations G37fs, G38fs and E248fs of the SOX10 gene. The constructs were verified by direct sequencing. NIH3T3 cells were transiently transfected with the expression plasmids of wide type SOX10, G37fs, G38fs and E248fs, respectively. The exogenous expression of WT SOX10 protein and mutant G37fs, G38fs and E248fs proteins were analyzed using Western blot assay, while their subcellular distribution were observed with an immunofluorescence assay.</p><p><b>RESULTS</b>The DNA sequences of expression plasmids for SOX10 and its mutant G37fs, G38fs and E248f were all correct. Both WT and mutant SOX10 proteins were detected at the expected site. WT SOX10 and E248fs proteins have only localized in the nucleus, whereas G37fs and G38fs proteins showed aberrant localization in both cytoplasm and nucleus.</p><p><b>CONCLUSION</b>Recombinant eukaryotic expression plasmids for the SOX10 gene and its mutants were successfully constructed. Preliminary analysis showed that the mutations have affected the subcellular distribution of WT SOX10 proteins, which has laid a basis for further study of the molecular mechanism of WS caused by SOX10 gene mutations.</p>

The expression and signi ficance of ING3 in colorectal cancer / 实用肿瘤学杂志

Objetcvi e Inhibitor of growth TNG3 is a candidate tumor suppressor gene ,which belongs to the human ING gene family .It has been interested by scientist because of its putative roles as tumor suppressors . There is a low expression level of ING3 in a variety of tumors.However,the relationship of ING3 and the develop-ment of colorectal cancer is unclear yet .The aim of this studyis to examine the expression of ING 3 in colorectal cancer by qRT-PCR.Methods Statistical analysis was performed with the expression of ING 3 mRNA which has been evaluated in 82 matched samples comprising primary colorectal cancer and paired non -cancerous mu-cosa samples.Results In 51%colorectal cancer patients ,the expression level of ING3 was significantly lower in cancer tissue than that in normal tissue .The expression level of ING3 was closely related to pT、pM stage , and TNM stage(P<0.05).The Univariate analysis showed that the pT、pN、pM stage,TNMstage and ING3 expression levels were associated with disease -free survival(P<0.05).CRC patients with low -expression level of ING3 showed shorter disease -free survivals(P<0.05).Conclusion The expression of ING3 might contribute to the CRC development or progression .

Generation and Expression of Recombinant Eukaryotic Expression Plasmids of PAX3 Gene and Its Significance / 听力学及言语疾病杂志

Objective To study exogenous expression and subcellular localization of wild type (WT ) and mu-tant PAX3 proteins in vitro by generating their expression plasmids for further study of pathogenesis of Waarden-burg syndrome (WS) .Methods The plasmids pECE-PAX3 and pcDNA3 .0-HA were ligased after they were cut by double enzyme digestion using molecular cloning technique to generate recombinant eukaryotic expression plasmid pcDNA3 .0-PAX3-HA ,which was as a template to generate expression plasmids pcDNA 3 .0 -H80D -HA and pcDNA3 .0-H186fs-HA of novel mutations H80D and H186fs of PAX3 gene .All constructs were verified by di-rect nucleotide sequencing .NIH3T3 cells were transfected transiently with the expression plasmids of PAX3 ,H80D and H186fs respectively .The exogenous expression of WT PAX3 protein and mutant H80D ,H186fs proteins were analysed using Western blot assay ,while their subcellular distribution were observed using immunofluorescence as-say .Results The DNA sequences of expression plasmids of PAX3 and its mutant H80D ,H186fs were correct . Both WT and mutant PAX3 proteins were detected at the expected size .WT PAX3 and H80D proteins were only lo-calized in the nucleus ,whereas H186fs protein showed aberrant localization in both cytoplasm and nucleus .Conclu-sion We successfully generated the recombinant eukaryotic expression plasmids of PAX 3 gene and its mutants and drew preliminary conlusion of gene mutation having effect on subcellular distribution of WT PAX 3 proteins in vitro , which lays experimental basis for further study of the moceluar mechanism of WS caused by PAX3 gene mutations in China .

Audiological Analysis of Patients with Intact Tympanic Membranes and Conductive or Mixed Hearing Loss / 听力学及言语疾病杂志

Objective To investigate audiological characteristics of patients with intact tympanic membranes and conductive or mixed hearing loss .Methods A retrospective study was carried out among 30 patients (42 ears) with intact tympanic membranes and conductive or mixed hearing loss who underwent exploratory tympanotomy . The preoperative outcomes of pure tone audiometry ,tympanometry ,resonant frequency of middle ear and temporal bone CT scan were analyzed .Results Among 42 ears ,30 ears with otosclerosis and 12 ears with ossicular chain dis-ruption were confirmed in exploratory tympanotomy ,but only 5 ears showed positive findings in CT scan .The mean thresholds of bone conduction ,air conduction and air -bone gap at frequencies of 0 .5 ,1 and 2 kHz were 27 .5 ± 1 .3 dB HL ,67 .0 ± 1 .8 dB HL ,39 .5 ± 1 .1 dB HL ,respectively .An analysis of tympanometric data of all patients re-vealed that 50% of all ears (21/42) were type A tympanograms ,42 .9% (18/42) were type As tympanograms ,and 7 .1% (3/42) were type Ad tympanograms .The mean of the resonant frequency of the middle ear in otosclerositic patients (1 079 .0 ± 67 .4 Hz) was significantly higher than ossicular chain disruption patients (633 .3 ± 43 .6 Hz) . Conclusion Otosclerosis is the most common in the patients with intact tympanic membranes and conductive or mixed hearing loss .The middle ear resonant frequency of otosclerositic patients is significantly higher than that of ossicular chain disruption patients .