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Objective:To study the risk factors of postoperative permanent hypoparathyroidism after total thyroidectomy.Methods:In this retrospective study ,the receiver operating characteristic (ROC) curve of postoperative 24 h parathyroid hormone(PTH) values was used to predict the risk of permanent hypoparathyroidism.The clinical and pathological features that related to postoperative permanent hypoparathyroidism were studied by χ2 test and multivariate Logistic regression analysis. Results:Eight hundred and eighty-nine patients were enrolled, the incidence of postoperative transient and permanent hypoparathyroidism was 33.3% and 4.0%, respectively. When 24 h PTH levels less than or equal to 5.84 pg/ml were used as cut off value, the sensitivity of the prediction of permanent hypoparathyroidism was 100%, the specificity was 72%, and the positive predictive value was 22.5%. The presence of parathyroid gland in the pathologic specimen, parathyroid autotransplantation and PTH≤5.84 pg/ml at 24 h after operation were statistically significant risk factor for permanent hypoparathyroidism in multivariate analysis( χ2=10.900, P=0.001; χ2=4.415, P=0.044; χ2=13.576, P=0.000). Group analysis found that the lower 24 h PTH the higher incidence of permanent hypoparathyroidism. Conclusions:For patients undergoing total thyroidectomy, when parathyroid gland is involved and/or the PTH value is low at 24 h after surgery, the occurrence of permanent hypoparathyroidism is high.
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Objective:To investigate the patterns of lateral and central neck lymph node metastasis in N1b papillary thyroid carcinoma (PTC) patients.Methods:From Jun 2018 to Sep 2020, 96 consecutive treatment-naive unilateral N1b PTC patients were included. After total thyroidectomy, bilateral central neck and unilateral lateral neck dissection (including the level Ⅱa, Ⅱb, Ⅲ, Ⅳ and Ⅴb) metastasis in different levels and pathological status of lymph nodes were analyzed.Results:The lymph node metastasis rates were 85% in ipsilateral paratracheal region, 82% in level Ⅲ, 77% in level Ⅳ, 55% in pre-tracheal region, 47% in contralateral paratracheal region, 40% in level Ⅱa, 34% in pre-laryngeal region, 6% in level Ⅱb, 6% in intermuscular region, 4% in level Ⅴb. Tumor size >1cm and pre-tracheal lymph nodes netastasis were associated with contralateral paratracheal metastasis ( OR=3.282 and 3.064, P<0.05); Metastasis to Ⅵa was associated with metastasis to Ⅵb ( OR=3.364, P<0.05). Conclusion:Level Ⅱa-Ⅳ of lateral neck and all subgroups of central neck lymph nodes tend to be involved in PTC with lateral neck metastasis. Lateral neck including level Ⅱa-Ⅳ and the whole central neck including right Ⅵb lymph nodes dissection is recommended for N1b PTC patients.
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Objective@#To investigate the diagnostic performance of fine-needle aspiration (FNA) cytology for the detection of lateral lymph node metastases in patients with papillary thyroid carcinoma (PTC).@*Methods@#A total of 109 lateral lymph nodes with suspicious metastases from 85 patients were retrospectively subjected to FNAC, fine-needle aspiration thyroglobulin measurement (FNATg), and FNATg/SerumTg measurement. Lymph node pathological results after surgery were taken as the gold standard. Using Mann-Whitney U test, Pearson linear model and ROC curve were used for data analysis.@*Results@#The sensitivity, specificity and accuracy of FNATg for the diagnosis of lateral neck lymph node metastasis were respectively 93.7%, 90.0% and 93.3% and those of FNATg/SerumTg were respectively 89.9%, 90.0% and 93.2% respectively, the threshold values for FNATg and FNATg/SerumTg were 0.925 ng/ml and 1.007, respectively. The sensitivity, specificity and accuracy of FNATg combined with FNAC were respectively 91.0%,93.5% and 94.4%. The existence of thyroid tissue and the expression of serum Tg did not affect the expression of lymph node FNATg. The FNATg cutoff value of 0.925 ng/ml showed the best diagnostic performance in patients with a thyroid gland, while the FNATg/SerumTg cutoff ratio of 14.95 showed the best diagnostic performance in patients without a thyroid gland. The serum TgAb significantly interfered with the expression of FNATg in the lateral neck metastatic lymph nodes (P=0.049).@*Conclusions@#FNATg alone or the combination of FNATg with FNAC are highly reliable in the diagnosis of lateral neck lymph node metastases in patients with PTC. The expression of TgAb may interfere with the accuracy of the diagnostic performance of FNATg.
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Objective@#To study the diagnostic value of a multi-gene molecular testing in cytologically indeterminate thyroid nodules.@*Methods@#From February 2018 to September 2018, patients with thyroid nodules who underwent fine needle aspiration(FNA) at Peking University Cancer Hospital were enrolled. Three hundred and sixty patients were included, consisting of 86 men and 274 women, with a mean age of 45.8 years (between 13 and 89 years old). Among 391 nodules, 141 were cytologically inderminate and 75 were resected. FNA samples underwent prospective testing using a next-generation sequencing (NGS) assay, which included 16 genes for point mutations and 26 types of gene fusions. The testing results of indeterminate nodules were compared with surgical outcomes, to determine the diagnostic performance. The results were compared with the BRAF V600E single gene mutation analysis by χ2 test.@*Results@#The multi-gene testing showed a sensitivity of 73.2%, specificity of 96.8%, positive predictive value of 96.8%, and negative predictive value of 73.2%. The diagnostic accuracy of multi-gene testing was significantly higher than the BRAF V600E mutation test (83.3% vs 73.6%, χ2=31.588, P<0.01).@*Conclusion@#Multi-gene testing in FNA samples is an effective method to diagnose cytologically indeterminate thyroid nodules, which has a higher accuracy than BRAF V600E mutation detection.
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Objective To explore the diagnostic value of conventional ultrasound(US) and contrast enhanced ultrasound(CEUS) in predicting extrathyroidal extension of papillary thyroid cancer(PTC).Methods Eighty-five PTCs in 75 patients were selected for thyroid surgery underwent ultrasound and contrast-enhanced ultrasound.The degrees of contact between PTCs and capsule were observed by US and CEUS respectively(0,0-25%,25%-50%,≥50%),and the diagnostic efficiency in different degree of contact (>0 %,≥25 %,≥50%) as preoperative diagnostic criteria were analyzed.The diagnostic efficiency between US and CEUS in predicting extrathyroidal extension of PTC were compared.Results Of the 85 PTCs,extrathyroidal extension was presented in 57 (67.06%) based on pathologic results.When the degree of contact (> 0 %,< 25 %,25 %-50 %,≥ 50 %) was gradually increased,the incidence of extrathyroidal extension of the thyroid cancer was also gradually risen (P <0.001).Comparing the sensitivity,accuracy,odds ratio,and Az value of three groups(>0%,≥25%,≥50%),it showed that the general diagnostic efficiency between two groups(>0%,≥25%) was similar by US and CEUS.However,the sensitivity and accuracy of >0% contact with the adjacent capsule were markedly higher than those of the other two groups(P <0.001).Selecting >0% contact with the adjacent capsule as preoperative criteria,the Az value of CEUS was markedly higher than that of US (Z =2.208,P =0.027).Conclusions The preoperative imaging feature of more than 0% contact with the adjacent capsule is more sensitive and accurate degree in predicting extrathyroidal extension of PTC.Compared with US,CEUS may serve as a better useful tool to predict extrathyroidal extension of PTC.
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To study the chemical constituents of Veratrum dahuricum (Turcz.) Loes. f., a new aurone glycoside named as (Z)-7, 4'-dimethoxy-6-hydroxyl-aurone-4-O-β-glucopyranoside was isolated from the 95% ethanol extracts of the rhizomes and roots of Veratrum dahuricum (Turcz.) Loes. f. by repeated column chromatography on silica gel and recrystallization. Its structure was established by extensive spectroscopic analyses, and its cytotoxicities against HepG-2, MCF7 and A549 cell lines were measured in vitro.
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Aim To investigate the roles of FFJ-5 in human breast cancer MCF7 cells and drug-resistant MCF7/DOX cells and to explore its mechanisms. Methods MTT assay was used to detect the effect of FFJ-5 on MCF7 and MCF7/DOX cell proliferation and sensitivity of doxorubicin in MCF7/DOX cells.West-ern blot was used to investigate the effect of FFJ-5 on expression of EGFR,p-EGFR,Akt,p-Akt,PKM2, cleaved caspase-3,cleaved PARP and P-gp.DNA lad-der analysis was performed to determine the effect of FFJ-5 on genomic DNA.RT-PCR was performed to de-tect the influence of FFJ-5 on multidrug resistance gene MDR1 mRNA levels.Results The results showed that FFJ-5 inhibited the growth of MCF7 ,inhibited the expression and activity of EGFR and Akt,and conse-quently reduced the expression of PKM2 in MCF7 cells;FFJ-5 activated caspase-3 and induced genomic DNA fragmentation;FFJ-5 also inhibited the growth of MCF7/DOX cells and enhanced the anti-tumor activity of doxorubicin in MCF7/DOX cells.Conclusion The results suggest that FFJ-5 could inhibit MCF7 cell growth and induce MCF7 cell apoptosis through inhibi-tion of EGFR-Akt-PKM2 pathway and activation of ap-optosis-related factors caspase-3 , meanwhile FFJ-5 could also reverse the resistance of MCF7/DOX.
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<p><b>OBJECTIVE</b>To evaluate the protection of parathyroid glands in bilateral papillary thyroid cancer patients who underwent total thyroidectomy and bilateral central compartment dissection.</p><p><b>METHODS</b>The clinical data of 78 patients undergoing total thyroidectomy and bilateral central compartment dissection between May 2013 and May 2014 were analyzed retrospectively.</p><p><b>RESULTS</b>Among 78 patients, 131 superior parathyroid glands were protected in situ, among those, 112 parathyroid glands located at the level of inferior edge of parathyroid cartilage; 19 parathyroid glands located in the superior 1/3 part of the back sides of the thyroid glands. All the superior parathyroid glands located in the superior and lateral-superior part of the 2 cm part down from the entrance of recurrent laryngeal nerve. A total of 110 inferior parathyroid glands were protected in situ, among those, 57 glands located in the superior of inferior thyroid artery; 24 glands located just on the surface of inferior thyroid artery; 29 glands located below the superior of inferior thyroid artery. Three parathyroid glands were found in the dissected tissues and were implanted immediately. Seven parathyroid glands were found with post-operative pathologic examinations. During surgery, four parathyroid glands was found in 27 patients, three in 30 patients, two in 16 patients, and one in 5 patients.</p><p><b>CONCLUSION</b>For papillary thyroid cancer patients who underwent total thyroidectomy and bilateral central compartment dissection, identification of parathyroid glands and protection of blood supply to glands are the most effective methods to prevent hypoparathyroidism.</p>
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Humans , Carcinoma , General Surgery , Carcinoma, Papillary , Hypoparathyroidism , Organ Sparing Treatments , Parathyroid Glands , General Surgery , Recurrent Laryngeal Nerve , Retrospective Studies , Thyroid Neoplasms , General Surgery , ThyroidectomyABSTRACT
Objective EGFR targeted therapy mediated by adeno-associated virus is a promising way to treat pancreatic cancer.This study aimed to assess the feasibility and activity of combining rAAV-anti EGFR,gemcitabine,and radiation in pancreatic cancer cells.Methods Aspc-1 human pancreatic carcinoma cells were divided into several groups,in vitro and in vivo,which were respectively exposed to gemcitabine alone,radiation alone,rAAV-anti EGFR alone,the combination of rAAV-anti EGFR with gemcitabine,the combination of rAAV-anti EGFR with radiation,and the combination of all three agents.The pancreatic cancer tumor growth and apoptotic rate were measured.Results The apoptotic rate was higher in cells treated with a single or combination of agents compared to the negative control (P<0.05).The combination of rAAV-EGFR,gemcitabine,and radiation produced the highest induction of apoptosis compared to a single agent alone (P < 0.05).Treatment with rAAV-anti EGFR greatly inhibited growth in the tumor xenografts (P<0.05),and a synergistic effect of rAAV-anti EGFR,gemcitabine,and radiation was found.The number of tissue cancer cells that expressed cleaved caspase-3 after treatment with rAAV EGFR was more than that of the control group (P<0.05).The combined treatment of rAAV-anti EGFR,gemcitabine,and radiation induced the highest numbers of cells expressing cleaved caspase-3 compared to that with a single agent alone (P<0.05).Conclusions The rAAV-anti EGFR therapy in combination with chemotherapy and radiation therapy demonstrated a greater efficacy over therapy with a single agent alone.rAAV-anti EGFR increased the efficacy of gemcitabine and radiation in the treatment of pancreatic cancer cells.
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This study is to investigate the inhibitory effect and mechanism of prosapogenin A (PSA) on MCF7. MTT assay was performed to determine the inhibitory effect of PSA on MCF7 cells. PI/Hoechst 33342 double staining was used to detect cell apoptosis. RT-PCR was used to test the mRNA levels of STAT3, GLUT1, HK and PFKL. Western blotting was performed to determine the expression of STAT3 and pSTAT3 protein in MCF7 cells. The results showed that PSA could dose-dependently inhibit cell growth of MCF7 followed by IC50 of 9.65 micrmol x L(-1) and promote cell apoptosis of MCF7. Reduced mRNA levels of STAT3, HK and PFKL were observed in MCF7 cells treated with 5 micromol x L(-1) of PSA. PSA also decreased the level of pSTAT3 protein. STAT3 siRNA caused decrease of mRNA of GLUT1, HK and PFKL which indicated STAT3 could regulate the expressions of GLUT1, HK and PFKL. The results suggested that PSA could inhibit cell growth and promote cell apoptosis of MCF7 via inhibition of STAT3 and glycometabolism-related gene.
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This study is to investigate the effect of small interfering RNA targeting STAT3 (STAT3-siRNA) enhancing antitumor activity of doxorubicin. RT-PCR and Western blotting were used to test the expression of STAT3 mRNA and protein in the HepG2, HeLa and K562/DOX cells and the effect of STAT3-siRNA on the expression of STAT3 mRNA and protein. MTT and Trypan blue assay were performed to determine the inhibitory effect of STAT3-siRNA on HepG2, HeLa and K562/DOX cells and the effect of STAT3-siRNA enhancing antitumor activity of doxorubicin. The effects of STAT3-siRNA on intracellular accumulation of doxorubicin and cell apoptosis were performed by Arrary Scan V(TI)HCS600 High-Contents. The results showed that STAT3 gene, STAT3 and pSTAT3 protein were highly expressed in HepG2, HeLa and K562/DOX cells and STAT3-siRNA decreased the expression of STAT3 mRNA and protein. STAT3-siRNA inhibited the growth of HepG2, HeLa and K562/DOX cells. STAT3-siRNA in combination with doxorubicin decreased by 3.13, 5.22 and 1.74 fold of IC50 of HepG2, HeLa and K562/DOX cells compared with doxorubicin only. Intracellular accumulation of doxorubicin increased by 16.8%, 12.87% and 25.67% respectively in HepG2, HeLa and K562/DOX cells in the presence of STAT3-siRNA. An enhancement of doxorubicin-induced cell apoptosis was observed in HepG2, HeLa and K562/DOX cells treated with STAT3-siRNA. The results suggested that STAT3-siRNA could enhance the antitumor activity of doxorubicin on HepG2, HeLa and K562/DOX cells.
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This study is to investigate the role of endogenous CSE/H2S in regulating apoptosis of HepG2 cells. MTT and Trypan blue assay were performed to determine the effect of CSE inhibitor PAG and CSE siRNA on proliferation of HepG2. Production of H2S from HepG2 cells was assessed spectrophotometrically using N, N-dimethyl-p-phenylenediamine-dihydrochloride. Cells apoptosis was detected by means of double staining of Hoechst 33342 and PI with Array Scan V(TI)HCS600 High-Contents. Dihydroethidine (DHE) and 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used to determine intracellular superoxide anion and ROS level. Reduced glutathione (GSH) was determined by OxiSelect Total Glutathione Assay Kit. Recombinant plasmid pcDNA 3.1/myc-His(-)-CSE was constructed and transfected into 293T cells to rescue the ROS and GSH level to further investigate the effect of CSE/H2S on ROS and GSH. Western blotting was performed to test the effect of CSE siRNA on expression of activated caspase 3 and p-AKT and Nrf2 protein. The results showed that PAG and CSE siRNA could significantly decrease the production of H2S in HepG2 cells and inhibit the proliferation of HepG2 cells at a dose-dependent and time-dependent manner, respectively. PAG and CSE siRNA could promote the cell apoptosis of HepG2 cells. Moreover, PAG and CSE siRNA induced increased ROS generation and depletion of the critical antioxidant GSH and recombinant plasmid pcDNA 3.1/myc-His(-)-CSE rescued the level of ROS and GSH. Meanwhile, CSE siRNA increased the expression of activated caspase 3, but CSE siRNA did not affect the expression of p-AKT and Nrf2. These results suggested that the CSE/H2S pathway was involved in suppression of HepG2 cell growth and promoted apoptosis of HepG2 cells in an oxidative stress-dependent manner.
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Pancreatic cancer is an aggressive malignant gastrointestinal tumor,surgical resection offers the only chance of cure. Due to its anatomic and biological characters,early stage pancreatic cancer is usually clinically silent,80%-85% of patients present with advanced unresectable disease. Furthermore,pancreatic cancer responds poorly to most radiochemotherapeutic agents.In this article,we discuss some clinical aspects including early diagnosis,preoperative systemic assessment,surgery and perioperartive adjuvant therapy,and we hope to find the challenges as well as strategies for pancreatic cancer in order to further improve the current level of diagnosis and treatment in China.
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Objective To construct a recombinant adeno-associated virus(rAAV)vector containing a human anti-epidermal growth factor receptor(anti-EGFR)single-chain variable fragment antibody gene,and observe its inhibitory effects on pancreatic cancer cell lines.Methods Human anti-EGFR single-chain variable fragment antibody gene was inserted into the Kpn I and Bgl Ⅱ sites to construct a rAAV-anti EGFR vector,and then rAAV1-EGFP group and rAAV1-anti EGFR group were established.The expression of anti-EGFR antibody was observed.Antibody expression was detected by Western blot,and the inhibition and apoptosis rates of human pancreatic cancer cell lines(PCT-3,SW1990,Capan-1,ASPC-1,MiaPaCa-2 and PANC-1 cells)were detected by CCK-8 assay and flow cytometry,respectively.All data were analyzed using the t test.Results The results of Western blot assay demonstrated that anti-EGFR antibody was expressed in 6 pancreatic cancer cell lines.The inhibition rates of rAAV1-EGFP and rAAVl-anti EGFR on pancreatic ASPC-1 cells were 1.1%± 2.4% and 15.1%±3.5%,respectively,with a significant difference between the 2 groups(t =6.598,P <0.05).The apoptosis rates of PANC-1 cells were 7.0% ± 3.0% in the rAAV1-EGFP group and 1 1.4% ± 2.5% in the rAAV1-anti EGFR group,with no significant difference between the 2 grouvs(t = 1.952,P >0.05).The apoptosis rates of SW1990,ASPC-1,Capan-1,PCT-3,MiaPaCa-2 cells were 1.1% ± 0.8%,1.5% ± 0.7%,1.7% ± 1.2%,1.1%±0.7% and 2.2% ± 1.1% in the rAAV1-EGFP group,and 17.6% ± 2.2%,46.9% ± 3.9%,20.0% ±2.8%,12.1% ± 1.6% and 31.1% ±2.5% in the rAAV1-anti EGFR group,respectively,with significant differences between the 2 groups(t = 12.208,19.846,10.405,10.909,18.327,P <0.05).Conclusions A rAAV-anti EGFR vector with human anti-EGFR single-chain variable fragment antibody gene was constructed.Anti-EGFR antibody has obvious inhibition effects on pancreatic cancer cell lines.
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The chemical constituents of Leonurus heterophyllus were separated and purified by repeated column chromatography on silica gel, HPD 100, Sephadex LH-20, and PHPLC. Each compound was characterized by spectroscopic and physical data. Eight compounds have been purified and identified to be quercetin 3-O-robinobioside (1), rutin (2), isoquerci trin (3), hyperoside (4), quercetin (5), apigenin (6), genkwanin (7), and benzoic acid (8). Among them, compounds 2, 5-7 were isolated from L. heterophyllus for the first time; Compounds 1, 3, 4, 8 were obtained for the first time from the genus Leonurus. The in vitro activities against leukemia K562 Cells of pure components were evaluated by testing their IC50. Compounds 1-6, 8 exhibited in-vitro inhibitory activities against leukemia K562 cells in different extent.
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Humans , Antineoplastic Agents , Chemistry , Pharmacology , Cell Proliferation , Drugs, Chinese Herbal , Chemistry , Pharmacology , K562 Cells , Leonurus , ChemistryABSTRACT
This study investigated the reversal effect of isotetrandrine, an isoquinoline alkaloid extracted from Caulis mahoniae, on P-glyeoprotein-mediated multidrug resistance in human breast cancer doxorubicin-resistant (MCF-7/DOX) cells. RT-PCR assay and immunity histochemistry assay were used to determine the expression level of mdrl gene and P-gp in MCF-7/DOX cells to elucidate resistant character of MCF-7/DOX cells. The activity of isotetrandine to enhance doxorubicin cytotoxicity was tested using MTT (3-(4,5-dimethyhhiazol)-2,5-diphenyltetrazolium bromide) assay and was evaluated by the reversal fold (RF) values. Intracellular accumulation of doxorubicin was assessed by the determination of doxorubicin-assoeiated fluorescence intensity. Effect of isotetrandrine on the expression level of P-gp in MCF-7/DOX cells was then determined by immunity histochemistry assay. The ability of isotetrandrine to inhibit P-gp function was evaluated by detecting the accumulation and efflux of rhodamine123 ( Rh123 )with flow cytometry (FCM). Verapamil was employed as a comparative agent in whole experiment. The results indicated that MCF-7/DOX cells had phenotype of MDR and that the positive expression of P-gp MCF-7/DOX cells and reversal fold (RF) was significantly higher than that of verapamil (P < 0.05 ), butit hardly affected cytotoxicity of DOX in MCF-7 cells and the expression level of P-gp in MCF-7/DOXcells. The ability of isotetrandrine to inhibit P-gp function was reversible, because incubation of MCF-7/DOX cells with isotetrandrine caused a marked increase in uptake and a notable decrease in efflux of Rh123 and a marked increase of intracellular DOX concentrations. In conclusion, isotetrandrine exhibited potent effect on the reversal of P-gp-mediated MDR in vitro, suggesting that it might become a candidate of effective MDR reversing agent in cancer chemotherapy.