Department of Clinicopathological analysis of papillary thyroid carcinoma with Hashimoto’s thyroiditis / 中华内分泌外科杂志

Objective:To discuss the effect of Hashimoto’s thyroiditis (HT) on papillary thyroid carcinoma (РТС) .Methods:The clinical features and pathological characteristics of 682 patients who underwent surgical treatment for the first time from Sep. 1st,2019 to May. 1st, 2021 in Department of Thyroid, Breast and Hernia Surgery, and confirmed by postoperative pathology as papillary thyroid carcinoma were retrospectively analyzed. There were 189 male patients, and 493 female patients, 529 patients < 55 years old and 153 patients ≥55 years old. 476 patients were classified as PTC group and 206 patients as PTC combined with HT group. Chi square test was used to compare the difference between two groups in gender, age, thyroglobulin antibody, thyroid stimulating hormone, thyroid peroxidase antibodies, thyroid peroxidase, number of lesions, metastasis lymph node in central region, thyroid stimulating hormone receptor antibody, carcinoembryonic antigen, whether microcarcinoma, vascular invasion, glandular outside violation, capsule and lateral transfer analysis, ultrasonic calcification, etc. At the same time, all patients were divided into the group without central lymph node metastasis (345 cases) and the group with central lymph node metastasis (337 cases) . The χ 2 test was used to compare the differences between the two groups in terms of sex, age, number of lesions, microcarcinoma, vascular invasion, extradular invasion, capsular invasion, lateral cervical lymph node metastasis, ultrasonic calcification and so on, so as to analyze the differences in clinical characteristics between the two groups. Results:There were 206 cases (30.21%) in PTC combined with HT group and 476 cases (69.79%) in PTC without HT group. There were significant differences in gender (12/194 vs 177/299) ( P=0.000) , age (175/31 vs 354/122) ( P=0.002) , TgAb (115/91 vs 455/21) ( P=0.000) ,TSH (13/175/18 vs 33/429/14) ( P=0.004) , TPOAb (90/116 vs 422/54) ( P=0.000) , number of lesions (114/92 vs 325/151) ( P=0.001) and lymph node metastasis in central area (87/119 vs 250/226) ( P=0.014) between the two groups ( P < 0.05) , but there were no significant differences in TRAb (196/10 vs 461/15) ( P=0.171) , CEA (205/1 vs 469/7) ( P=0.478) , microcarcinoma (136/70 vs 309/167) ( P=0.781) , vascular invasion (4/202 vs 16/460) ( P=0.446) , extraglandular invasion (52/154 vs 108/368) ( P=0.470) , capsule invasion (149/57 vs 358/118) ( P=0.429) , lateral neck lymph node metastasis (31/175 vs 72/404) ( P=0.979) or ultrasonic calcification (157/49 vs 392/84) ( P=0.063) . Compared with PTC group, PTC combined with HT group had the characteristics of more women, younger age, high TgAb, high TSH, high TPOAb, multiple lesions and high proportion of non central lymph node metastasis. There were 345 cases (50.59%) without central lymph node metastasis and 337 cases (49.41%) with central lymph node metastasis. Gender (71/274 vs 118/219) ( P=0.000) , age (246/99 vs 283/54) ( P=0.000) , exadular invasion (66/279 vs 94/243) ( P=0.007) , number of lesions (240/105 vs 199/138) ( P=0.004) , microcarcinoma (259/86 vs 186/151) ( P=0.000) , calcification on ultrasound (250/95 vs 299/38) ( P=0.000) , and HT (119/226 vs 87/250) ) ( P=0.014) had statistical significance ( P<0.05) but had no statistical significance in capsule invasion (250/95 vs 257/80) ( P=0.256) or vascular invasion (10/335 vs 10/327) ( P=0.958) . In addition, patients in the group with central lymph node metastasis were more male, younger, with multiple lesions, exadenocarcinoma, less microcarcinoma, and calcification on ultrasound without hashimoto. Univariate analysis showed that gender, age, number of lesions, extraglandular invasion, calcification, microcarcinoma and Hashimoto had significant effects on lymph node metastasis in the central region; Multivariate analysis showed that the presence of microcarcinoma, ultrasonic calcification, Hashimoto and the number of lesions were independent risk factors for central lymph node metastasis. Conclusion:HT may promote the occurrence of PTC, but at the same time inhibit its development, so that PC patients with HT have a better prognosis.

Ferroptosis-related gene and its relationship with prognosis of differentiated thyroid carcinoma patients / 中华内分泌外科杂志

Objective:To analyze the marker of ferroptosis-related genes in differentiated thyroid carcinoma (DTC) based on TCGA database.Methods:The mRNA expression profiles and survival information of thyroid cancer patients and normal thyroid samples were downloaded from the TCGA database. The genetic difference analysis added 653 normal thyroid samples from the GETx database. Twenty-two ferroptosis-related genes were selected for univariate Cox regression analysis. Genes associated with prognostic in the TCGA cohort were further screened and prognostic models using LASSO regression was constructed. Adjusted P<0.05 and | log2FC>1" as the threshold, 22 differentially expressed genes were selected. The genes were screened by multivariate Cox regression analysis for prognosis-related genes and displayed in a line diagram. Results:22 of the 24 ferroptosis-related genes were differentially expressed between the tumor and normal tissues, with13 high expression, 9 low expression, 1 gene expression without difference and 1 gene not expressed in half of the samples. Univariate Cox regression analysis found that DPP4 and TFRC were associated with the degree of disease risk (HR was<1 and>1, respectively) . When integrating GPX4, TFRC and DPP4 into the LASSO model screening, it was found to be related to prognosis after dividing the patients into risk groups according to lambda. min=0.0027, Riskscore= (0.7316) * TFRC+ (-0.2539) *DPP4 (Log rank P=0.00635. Multivariate Cox regression analysis found that DPP4 and TFRC were related to the degree of disease risk (HR were<1 and>1, respectively) . Conclusion:The model of TFRC and DPP4 constructed by ferroptosis-related differential expression genes may be potential predictive markers of DTC patients, which still needs further verification and will provide theoretical basis for further clinical treatment.

One case of recurrent parathyroid carcinoma complicated with lung metastasis after parathyroid adenoma operation and literature review / 中华内分泌外科杂志

This paper reports the clinical data of a patient with recurrent metastatic parathyroid carcinoma. The causes, clinical manifestation, diagnose, treatment and prognosis of parathyroid carcinoma were discussed in order to perfect the experience of diagnosis and treatment and improve the survival rate of such patients.

Effect of annexin A1 on the proliferation, migration and invasion of papillary thyroid carcinoma / 中华内分泌外科杂志

Objective:To investigate the effect of ANXA on biological behavior of papillary thyroid carcinoma (PTC) cells by interfering with the expression of annexin A1 (ANXA1) in PTC cell lines by short hairpin RNA (shRNA) .Methods:The shRNA with specific and high efficiency was designed to specifically interfere with the expression of ANXA1 in TPC-1 and BCPAP cell lines, and transfect the TPC-1 and BCPAP cell lines respectively, including specific ANXA1 interference and negative control virus transfection, and they were divided into shANXA1 group and negative control virus group. Semi-quantitative reverse transcription PCR (Q-PCR) and Western Blot were employed to verify gene expression. The shANXA1 group was used as the experimental group, the untransfected virus group and the negative control virus group were set as the control groups. The expression levels of ANXA1 in the three groups were compared and the shRNA interference efficiency was verified. The effects of ANXA1 knockdown on the proliferation, migration and invasion of TPC-1 and BCPAP cell lines were investigated by scratch, CCK8 and Transwell invasion experiments. Independent sample t test was used to compare the means between the two groups, and one-way analysis of variance was employed to compare multiple groups, with P<0.05 as statistically significant. Results:shRNA could efficiently silence the expression of ANXA1 at the transcription and translation level in PTC cell lines. Compared with the negative control cells, the cells proliferated after successful lentiviral transfection of TPC-1 and BCPAP (BCPAP, 24h: F= 25.15, P<0.001; 48h: F=6.44, P<0.001; 48h: F=46.94, P<0.001; TPC-1, 24h: F=207.50, P<0.001; 48h: F=202.45, P<0.001; 48h: F=55.89, P<0.001) , its migration (BCPAP, F=12511.10, P<0.001; TPC-1, F=3966.10, P<0.001) and invasion ability (BC-PAP: F=94.65, P<0.001; TPC-1: F=681.74, P<0.001) significantly decreased. Conclusion:After shRNA knock-down of ANXA1 gene, the proliferation, migration and invasion ability of TPC-1 and BCPAP cell lines decreased significantly, indicating that silencing this gene can reduce tumor aggressiveness, and initially reveals that ANXA1 may be an important potential in PTC biotherapy Target.

Clinical diagnosis and treatment of primary thyroid lymphoma: 19 cases / 中华内分泌外科杂志

Primary thyroid lymphoma is rare in the population, and with the improvement of the diagnostic efficiency of the needle biopsy technique, the effect of surgical treatment of primary thyroid lymphoma is reduced, but the surgical biopsy is still irreplaceable when the pathological subtype is not clear. The prognosis of primary thyroid lymphoma is mainly related to the pathological type and the stage of the disease.

Progress in diagnosis and treatment of Riedel's thyroiditis / 中华内分泌外科杂志

Riedel's thyroiditis (RT) is a rare form of thyroid disease,and the rate of misdiagnosis is high due to the low incidence of RT and the lack of understanding of the disease by clinicians.There is currently no standard and guidance for the diagnosis and treatment of RT.Improving the understanding of RT is of great significance for standardizing diagnosis and treatment and improving the life quality of patients.

MicroRNA-16 inhibits cell proliferation and migration by targeting heat shock protein 70 in heat-denatured dermal fibroblasts

BACKGROUND/AIMS@#This study aimed to investigate the precise mechanism and function of miR-16 in heat-denatured primary human dermal fibroblasts.@*METHODS@#Primary human dermal fibroblasts were separated from normal human skin samples. Under heat stress, the levels of miR-16 and heat shock protein 70 (HSP70) were detected in primary human dermal fibroblasts by quantitative real-time polymerase chain reaction (qRT-PCR). Next, heat-denatured cells were transfected with synthetic scrambled negative control (NC) RNA (NC group), miR-16 mimics, miR-16 inhibitor or miR-16 inhibitor accompanied by small interfering RNA targeting HSP70, then the mRNA level of HSP70 was detected by qRT-PCR, cell proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and AlamarBlue assay, cell migration was examined by Transwell assay and cell apoptosis was assessed by transferase dUTP (deoxyuridine triphosphate) nick end labeling (TUNEL) assay. In addition, cell apoptosis-related proteins, Bax and Bcl-2, were detected by Western blotting.@*RESULTS@#Heat stress significantly reduced miR-16 level and increased the mRNA level of HSP70 compared with untreated cells (p < 0.05). Overexpressed miR-16 reduced the mRNA level of HSP70, suppressed cell proliferation (p < 0.05 or p < 0.01), migration (p < 0.05), and promoted cell apoptosis (p < 0.001) compared with the NC group. Down-regulated miR-16 exerted an opposite effect on primary human dermal fibroblasts with heat-denaturation. Furthermore, effects of miR16 down-regulation on cell proliferation and migration were reversed by HSP70 silence.@*CONCLUSIONS@#MiR-16 might have an inhibitory effect on cell proliferation and migration in heat-denatured human dermal fibroblasts, and HSP70 might be associated with the cell proliferation and migration as a target gene of miR-16.

The necessity and strategy of multidisciplinary team in management of thyroid carcinoma / 中华内分泌外科杂志

Thyroid carcinoma is the most common malignancy in the endocrine system.Most patients cannot be cured with operation alone.The concept and implementation of multidisciplinary team (MDT) in clinical oncology attributes to improved diagnosis and treatment of cancer.MDT plays an important role in diagnosis,treatment,and post operational management of thyroid carcinoma,as well as in the application of novel techniques and translational medicine.MDT can maximize the expertise of various disciplines,strengthen inter disciplinary cooperation,and provide standardized and individualized comprehensive treatment for patients with thyroid cancer.The most important benefit of MDT is that individual patient gets the most appropriate treatment decision made by a team of experts,including endocrinologists,nuclear medicine specialists,pathologists,radiologists,radiation therapists,and surgeons.This will be of great significance to improve quality of life and prognosis,at the same time,avoid over-treatment of thyroid cancer.

Expression of Pendrin gene (SLC26A4) and protein in multinodular goiter / 中华内分泌外科杂志

Objective To explore the expression of the Pendrin gene (SLC26A4) and protein in multinodular goiter.Methods Thyroid tissues were obtained from 40 multinodular goiter patients undergoing surgery while the control group were obtained from 40 nomal thyroid tissues.RT-PCR was used to test SLC26A4 gene while western blot and immunohistochemistry were used to test Pendrin protein expression and distribution.Results SLC26A4 mRNA expression in multinodular goiter tissue was significantly increased in comparison with normal nodular tissues (t=2.663,P=0.011).Pendrin protein expression in multinodular goiter group was higher than that in normal tissue (t=2.286,P=0.026).The immunohistochemistry results showed that the Pendrin protein in multinodular goiter was mainly located in cytoplasm.There was positive expression in 24 patients (60％) in multinodular goiter group,while it was in 14 patients (35％) in the normal control group.The difference was significant (X2=5.013,P=0.025).Pendrin protein mainly expressed in cytoplasm in multinodular goiter tissue while it was mainly in cytomembrane in the normal control group.Conclusion SLC26A4 mRNA and its coding protein Pendrin expression are increased in multinodular goiter group,and mainly located in cytoplasm,indicating that iodide transporter function may be damaged when multinodular goiter occurs.

Inhibitive effect of IL-35 on cardiac allograft rejection in mice / 中华器官移植杂志

Objective To investigate the action mechanism of IL-35 gene transfection ameliorating cardiac allograft rejection and prolonging allograft survival.Methods pEBI3-L-p35-Fc plasmid was amplified by polymerase chain reaction.In vitro plasmid DNA pEBI3-L-p35-Fc or pSec-L-Fc was,respectively,transfected into HEK293 cells using Lipofectamine 3000.At 48 and 72 h after transfection,IL-35 concentration in culture supernatant of transfected HEK293 cells was detected by ELISA.Balb/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of IL-35,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with IL-35 plasmid (50μg) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.The IL-35 expression in the blood,CD4+ CD25+ Tregs proportion in the blood and spleen,and the survival and the histopathologic changes of the cardiac grafts were also observed.Results In vitro the transfected HEK293 cells expressed IL-35.IL-35 enhanced the proliferation of CD4+ CD25+ Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (16 days) was significantly longer than in control group (7 days) (P＜0.01).As compared with control group,CD4+ CD25+ Tregs proportion was significantly increased (P＜0.01),CD8+ T cells proportion was decreased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited in the experimental group.Conclusion IL-35 could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25+ Tregs and inhibition of proliferation of CD8 + effector T cells.

Expressions of miRNA-21 and PTEN and the clinical significance in colorectal carcinoma / 中华普通外科杂志

Objective To explore the correlations between expression of miRNA-21 and PTEN and the invasion, metastasis of colorectal carcinoma.Methods The mRNA level of miRNA-21 and mRNA and protein levels of PTEN were assayed by real-time quantitative PCR and immunohistochemical methods respectively in 65 colorectal carcinoma specimen.Results The expression of miRNA-21 mRNA in cancer tissues was significantly higher than that in paracancerous tissues (t =3.50, P ＜ 0.05).Both mRNA and protein expressions of PTEN in tumor tissue were significantly lower than that in paracancerous tissues (t =7.35,t =12.23;P ＜ 0.05).Expression of miRNA-21 mRNA, PTEN mRNA and protein were obviously related with depth of invasion(F =18.36 ,F =17.26 ,F =12.83;P ＜ 0.05), Dukes stage (F =31.25, F =24.43, F =57.12;P ＜ 0.05) and lymph node metastasis (t =5.85, t =2.18, t =4.05;P ＜ 0.05), while it was not related to tumor differenciation and tumor location (F =7.39, t =4.62;F =7.78, t =1.29;F =5.14,t =1.37;P ＞ 0.05).Positive relation between miRNA-21 and PTEN in colorectal carcinoma was identified (r =-0.994, r =-0.927;P ＜ 0.05).Conclusion High expression of miRNA-21 and low expression of PTEN are both closely associated with invasion and metastasis of colorectal carcinoma.

Research progress of contrast-enhanced ultrasound and its imaging characteristics / 国际外科学杂志

As a technique which can assess both the micro-vascularization and macro-vascularization of the tumor,contrast-enhanced ultrasound have been developed rapidly,and its researches in thyroid nodules check and differentiation has became more and more widely.Different infusion modes in the diagnostic of thyroid nodules are really meaningful.The development of CEUS and identification of thyroid nodules will be reviewed in this article.

Preoperative serum alkaline phosphatase: a predictive factor for early hypocalcaemia following parathyroidectomy of primary hyperparathyroidism / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Postoperative hypocalcemia is one of the most common complications following parathyroidectomy for primary hyperparathyroidism (PHPT). The aim of this study was to analyze the predictive value of biochemical parameters as indicators for episodes of hypocalcemia in patients undergoing parathyroidectomy for PHPT.</p><p><b>METHODS</b>The patients with PHPT who underwent parathyroidectomy between February 2004 and February 2014 were studied retrospectively at a single medical center. The patients were divided into biochemical, clinical, and no postoperative hypocalcemia groups, based on different clinical manifestations. Potential risk factors for postoperative hypocalcemia were identified and investigated by univariate and multivariate Logistic regression analysis.</p><p><b>RESULTS</b>Of the 139 cases, 25 patients (18.0%) were diagnosed with postoperative hypocalcemia according to the traditional criterion. Univariate analysis revealed only alkaline phosphatase (ALP) and the small area under the curve (AUC) of receiver operating characteristics (ROC) curve for ALP demonstrates low accuracy in predicting the occurrence of postoperative hypocalcemia. Based on new criteria, 22 patients were added to the postoperative hypocalcemia group and similar biochemical parameters were compared. The serum ALP was a significant independent risk factor for postoperative hypocalcemia (P = 0.000) and its AUC of ROC curve was 0.783. The optimal cutoff point was 269 U/L and the sensitivity and specificity for prediction were 89.2% and 64.3%, respectively.</p><p><b>CONCLUSIONS</b>The risk of postoperative hypocalcemia after parathyroidectomy should be emphasized for patients with typical symptoms of hypocalcemia despite their serum calcium level is in normal or a little higher range. Serum ALP is a predictive factor for the occurrence of postoperative hypocalcemia.</p>

Review the Function and Structure of Interleukin-35 and Its Function in Mediating Treg Cell Immuno-Suppression / 天津医药

Interleukin (IL)-35 is a new member of IL-12 family who is secreted by T regulatory cells to strongly sup?press immuno reaction directly or indirectly. There is a growing interest in the using IL-35 as a potential therapy for chronic in?flammatory , cancer and autoimmune diseases. Here we reviewed IL-35 and its function as a mediator of suppression.

Diagnostic value of thyroid imaging reporting and data system in thyroid cancer / 中国肿瘤临床

Objective: To retrospectively characterize and analyze the thyroid ultrasound images of thyroid cancer patients, to classify thyroid nodules according to thyroid imaging reporting and data system (TI-RADS), and to evaluate the diagnostic value of TI-RADS. Methods:Clinical data of 160 thyroid cancer patients with 235 nodules from September 2008 to August 2011 were retro-spectively analyzed. All patients underwent conventional color Doppler ultrasonography to evaluate the size, number, shape, boundary, and extent of lesions and blood distribution of the nodules. All nodules were analyzed according to TI-RADS classification and scored to evaluate the diagnostic value of TI-RADS. Results:Among 176 malignant nodules, 144 (81.8%) had lesions showing a mass of irreg-ular boundary with no envelope and halo, 152 (86.4%) were inhomogeneous hypoechoic or slightly hypoechoic, or 144 (81.8%) exhibit-ed microcalcification. TI-RADS scores of 4 or 5 and 2 or 3 were obtained in 160 (90.91%) and 16 (9.09%) of the malignant nodules, re-spectively. Conclusion:Thyroid ultrasound sonogram of ill-defined, irregular shape, hypoechoic or slightly hypoechoic mass, microcal-cification within the mass, and rich blood flow is an important index for the diagnosis of thyroid cancer. Thyroid nodules with a TI-RADS score 4 or 5 have higher possibility for cancer. Therefore, TI-RADS has a significant diagnostic value for thyroid cancer be-fore operation.

Prolongation of cardiac allograft survival in mice by Decitabine-induced upregulation of Foxp3 expression / 中华器官移植杂志

Objective To investigate the effect of Foxp3 gene expression induced with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (Aza) in recipients on ameliorating allograft rejection and prolonging allograft survival and the possible mechanisms.Method BALB/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of Aza,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with decitabine (1.5 mg/kg/day) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.CD4+ CD25+ Tregs proportion in the blood and Foxp3 mRNA in cardiac grafts and spleen of recipients were measured respectively,and the survival and the histopathologic changes of the cardiac grafts were also observed.Result Decitabine enhanced the proliferation CD4 + CD25 + Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (11 days) was longer than that in control group (7 days) (P＜0.01).As compared with control group,Foxp3 mRNA expression in the cardiac grafts and spleen in recipient mice was significantly up-regulated,and CD4+ CD25 + Tregs proportion was increased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited.Conclusion Up-regulation of Foxp3 gene expression induced with decitabine in recipients could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25 + Foxp3 + Tregs.

Surgical therapy for malignancy patients complicated with hyperthyroidism / 中华普通外科杂志

Objective To summarize the therapeutic experience on malignancy patients complicated with hyperthyroidism.Methods Clinical date of 10 cases of surgical malignancy complicated with hyperthyroidism admitted from May 2004 to May 2010 in Tianjin Medical University General Hospital were analyzed retrospectively.Results Four cases were treated by subtotal thyroidectomy before radical operation for cancer.Radical operation for cancer was performed on 6 patients after clinical symptoms of hyperthyroidism were controlled by perioperative antithyroid agents.Postoperatively 3 patients complicated with clinical manifestations similar to thyroid crisis.There was no postoperative mortality.Chemotherapy was given to 10 patients,and 1 patient was discontinued for chemotherapy caused leucopenia.Conclusions Hyperthyroidism should be controlled by surgery or antithyroid agents before patients of malignant diseases could proceed with radical surgery.Proper preoperative medication and effective postoperative management can reduce operation risk and help the patients get through the perioperative period safely.

Lentivirus-mediated RNAi against xenoantigen in mouse vascular endothelial cells / 中华器官移植杂志

Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.

The inhibiting effect on rat C Ⅱ TA and MHC Ⅱ gene expression by plasmid vector-based RNAi technulogy / 中华微生物学和免疫学杂志

Objective To investigate the inhibitory effect on rat MHC Ⅱ transactivator (C Ⅱ TA) and MHC Ⅱ gene expression by plasmid vector-based RNAi technology. Methods According to C Ⅱ TA ge-netic information, three short hairpin RNA (shRNA) sequences were designed and the corresponding plas- mid vectors were constructed. Rat dendritic cell (DC) was transfected in vitro and rat spleen was transfected in vivo. Real time RT-PCR and flow cytometry were used to detect the expression of C Ⅱ TA and MHC Ⅱ in DC and spleen after transfection. Results C Ⅱ TA-shRNA plasmids were successfully constructed. Com-pared with control groups, the mRNA transcription levels of C Ⅱ TA and MHC Ⅱ and the expression level of MHC H were significantly inhibited in all three pC Ⅱ TA-shRNA experimental groups ( P < 0.01 ). There was positive correlation between the expression of C Ⅱ TA and MHC Ⅱ. Among the three shRNA groups, the first one showed the strongest inhibitory effect. Conclusion The expression of rat C Ⅱ TA and MHC Ⅱ can be obviously inhibited by plasmid vector of shRNA targeting C Ⅱ TA gene, which can be used for further investi-gation of gene therapy.

Aminoglycosides Modifying Enzyme Genes of Acinetobacter baumannii in Burn Unit / 中华医院感染学杂志

OBJECTIVE To study the aminoglycosides modifying enzyme genes and detect resistance of the Acinetobacter baumannii in burn unit.METHODS The susceptibility of the A.baumannii strains to 11 antibiotics were tested by K-B method.The aminoglycosides modifying enzyme genes were analyzed using polymerase chain reaction(PCR).RESULTS The resistance rate to antibacterials was as follows:to TZP 61.85%,SCF 23.69%,CAZ 64.48%,FEP 63.17%,IPM 63.17%,AK 56.58%,and to CIP 73.69%.In 76 strains of A.baumannii,48 strains(63.16%) were with aac(3)-Ⅰ,39(51.32%) with aac(6′)-Ⅰ,and 46(60.53%) with ant(3″)-Ⅰ,others were negative.The total test rate of aminoglycosides modifying enzyme gene was 63.16%.CONCLUSIONS There are not only multi-drug-resistance but also 63.16% aminoglycosides modifying enzyme genes of the A.baumannii isolated from burn unit.