Recurrent laryngeal nerve inlet zone lymph node metastasis in papillary thyroid cancer / 中华普通外科杂志

Objective:To explore the clinical significance of recurrent laryngeal nerve inlet zone(RLNIZ) lymph node metastasis in papillary thyroid cancer(PTC).Methods:The clinical data of the clinicopathologic characteristics of 738 cases with papillary thyroid cancer at our centers from Jul 2017 to Jun 2018 was retrospectively reviewed. 108 cases with RLNIZ lymph node dissection for pathological examination were included. The relationship between metastasis of RLNIZ lymph node and clinicopathologic characteristics was analyzed.Results:RLNIZ lymph node was detected in 12.3%(91/738)cases, the mean lymph node number in RLNIZ was 1.5±0.7, and 30.8%(28/91) cases suffered RLNIZ lymph node metastasis. RLNIZ lymph node metastasis(LNM) is associated with tumor size( P=0.028), capsular invasion( P=0.019), No. of central compartment LNM( P<0.001) and lateral neck LNM( P<0.001). No. of central compartment LNM was found to be the independent risk factor of RLNIZ lymph node metastasis. The incidence of dysphagia and inferior parathyroid damage was 0.9%(1/108)respectively. Conclusions:RLNIZ lymph node metastasis is common among PTC patients , therefore, RLNIZ lymph node should be routinely removed especially in patients with tumor size over 1cm、suspected capsular invasion and lateral neck lymph node metastasis confirmed by preoperative imaging examination.

Priliminary study of the effect of tumor-associated fibroblasts on the infiltration and metastasis of papillary thyroid carcinoma / 中华内分泌外科杂志

Objective:To detect the expression of fibroblast-specific protein 1 (FSP1/S100A4) , ɑ-smooth-muscle actin (ɑ-SMA) and fibroblast-activated protein (FAP-ɑ) in tumor-associated fibroblasts (TAFs) in papillary thyroid carcinoma (PTC) , and to investigate its relationship with the origination and development of PTC.Methods:The expression of FSP1/S100A4, ɑ-SMA and FAP-ɑ in normal thyroid and PTC was determined by SP method of immunohistochemistry, and the relationship between these indicators and important clinicopathological parameters were analyzed.Results:The positive expression of FSP1/S100A4, ɑ-SMA and FAP-ɑ was observed in PTC, but not detected in the follicular epithelium or stromal cells of normal thyroid. In addition, the expression of FAP-ɑ was significantly related to tumor size, lymph node metastasis and TNM classification ( χ2=6.833, P<0.05; χ2=10.296, P<0.05; χ2=4.910, P<0.05) . The expression of ɑ-SMA was positively related to the invasion of capsule and lymph node metastasis ( χ2=6.008, P<0.05; χ2=11.766, P<0.05) . The expression of FSP1/S100A4 was negatively related to the clinicopathological parameters above ( P>0.05) in PTC. Conclusion:TAFs in PTC may indicate the infiltration and metastasis, which provideds new thinking for the treatment strategies of papillary thyroid carcinoma.

The expression and clinical significance of miR-143-3p in papillary thyroid cancer / 中华内分泌外科杂志

Objective:To explore the expression and clinical significance of miR-143 in papillary thyroid cancer (PTC) .Methods:Tumor samples and adjacent tissues from 52 patients with PTC were obtained from Jan. 1st, 2018 to Mar. 31st, 2018 in Thyroid Surgery Department of the Affiliated Yantai Yuhuangding Hospital of Qingdao University. Quantitative reverse-transcriptase PCR (RT-qPCR) was used to measure the expression of miR-143 in those subjects. In addition, the relationship between the expression levels of miR-143 and the clinicopathological characteristics was analyzed.Results:RT-qPCR indicated that the expression of miR-143 was down-regulated in PTC, which was significantly lower than that in adjacent tissues ( t=-21.39, 95% CI: 18.20-15.07, P<0.001) . Low expression of miR-143 was related to the number of lymph node metastasis ≥3 in central compartment ( t=10.13, P=0.012) and lateral neck lymph node metastasis ( t=-4.67, P<0.001) . Conclusion:Downregulation of miR-143 in PTC is linked to the metastasis of PTC and may be a potential target for therapeutic intervention.

Clinical significance of monitoring drainage fluid parathyroid hormone after thyroid surgery / 中华内分泌外科杂志

Objective To explore the clinical significance of monitoring drainage fluid parathyroid hormone (dPTH) for estimating the in situ reserves and function of the parathyroid by analyzing the change of serum calcium,serum parathyroid hormone(sPTH) and dPTH after thyroid surgery.Methods According to the operative method,the total of 144 patients with thyroid disease were divided into five groups:unilateral lobectomy,unilateral lobectomy plus isthmectomy with unilateral lymph node dissection,total thyroidectomy,total thyroidectomy with unilateral lymph node dissection,and total thyroidectomy with bilateral lymph node dissection group.The blood calcium,sPTH and dPTH level of patients were tested before operation and on the 1st,2nd,3rd and 4th day after operation.The depression of serum calcium,hypocalcemia and hypoparathyroidism were observed after operation.The serum calcium,serum PTH and dPTH level were summarized and analyzed statistically in order to evaluate the in situ reserves and postoperative function of the parathyroid.Results Among the 114 cases,the decline of serum calcium level mostly happened on the 2nd day after operation(70 cases,61.4％).There were 36 patients with hypocalcemia (31.58％) and 34 patients with hypoparathyroidism (29.82％).Serum calcium level increased gradually in all of the patients.Although sPTH level swung,it had a rising trend on the whole.The level of serum calcium and sPTH was positively correlated.The level of dPTH was discrete and decreased along with time.The decline level of dPTH among different groups had statistical difference.Conclusions It is a promising method to evaluate the in situ reserves and function of the parathyroid by monitoring the level and changes of dPTH after thyroid surgery,and it is of value for preventive calcium supplementation after thyroid surgery.

Application of Dexmedetomidine for Endovascular Stent Graft Exclusion for Aortic Dissection / 中国微创外科杂志

Objective To evaluate the efficacy and safety of dexmedetomidine applied in the endovascular stent graft exclusion for aortic dissection . Methods From January 2015 to December 2016, 82 patients with type DeBakey Ⅲ aortic dissection in our hospital were divided into either experimental group or control group according to random number table .In the experimental group, patients were given 15 min infusion of dexmedetomidine at 1 μg/kg, followed by 0.3 μg· kg-1 · h-1 continuous infusion .In the control group , patients were given propofol infusion at 4-6μg· kg-1 · h-1 combined with target controlled infusion of remifentanil at 3-5 μg/L.The two groups were operated under local anesthesia .The Ramsay score, MAP, and SpO2 were observed at baseline time ( T0 ) , anesthesia induction ( T1 ) , skin cut ( T2 ) , and accomplishment of surgery ( T3 ) . Results The Ramsay scores of the experimental group were lower than those of the control group at T 0, T1, T2, and T3[T0:(1.8 ±0.4) points vs.(2.1 ±0.6) points, t=2.664, P=0.009;T1:(2.8 ±0.3) points vs.(5.1 ±0.4) points, t=29.454, P=0.000;T2:(3.0 ±0.5) points vs.(4.8 ± 0.3) points, t=19.766, P=0.000; T3: (2.9 ±0.4) points vs.(3.9 ±0.6) points, t=8.880, P=0.000].The MAP of the experimental group was higher than that of the control group at T 1[(76.2 ±10.1) mm Hg vs.(67.2 ±11.4) mm Hg, t=-3.784, P=0.002],and there was no significant difference at other timepoints between the two groups (P >0.05).The SpO2 of the experimental group at T0, T1, and T2 were higher than those of the control group [T0: (96.0 ±1.0)% vs.(95.0 ±1.5)%, t=-3.552, P=0.000;T1:(95.5 ±1.1)% vs.(90.0 ±1.5)%, t=-18.933, P=0.000; T2: (95.0 ±1.3)% vs.(90.8 ± 1.1)%, t=-15.792, P=0.000].There was no significant difference in SpO 2 at T3 between the two groups(t=0.220,P=0.827).Conclusion Dexmedetomidine can be used safely in aortic dissection stent graft exclusion .

Clinical significance of lymphatic metastasis between sternocleidomastoid and sternohyoid muscle in papillary thyroid cancer / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the clinical significance of metastasis of lymph nodes between sternocleidomastoid and sternohyoid muscle (LNSS) in papillary thyroid cancer (PTC).@*Methods@#A total of 175 patients with PTC who underwent thyroidectomy with LNSS dissection were retrospectively analyzed. Univariate and multivariate Logistic regression analyses were used to determine the independent risk factors for LNSS metastasis in PTC.@*Results@#The rate of detectable LNSS was 70.9% (124/175) and metastasis rate was 7.4% (13/175). Of 13 cases with LNSS metastasis, 10 with the coexistence of cervical lymph node metastasis. Univariate Logistic regression analysis showed that multiple focal cancer, tumor located in the lower pole of thyroid, belt-shaped muscle invasion, lateral cervical lymph node metastasis, cN+ , the number of cervical lymph nodes with metastasis and the number of lymph nodes with metastasis in level Ⅳwere the risk factors for LNSS metastasis (P<0.05). Multivariate Logistic regression analysis suggested that tumor located in the lower pole of thyroid and the number of cervical lymph nodes with metastasis >6 were the independent risk factors for LNSS metastasis (P<0.05). Given the number of cervical lymph nodes with metastasis as a predictor for the LNSS metastasis, the sensitivity was 92.3%, the specificity was 66.7% and the accuracy rate was 68.6%.@*Conclusions@#LNSS metastasis is commom in PTC, with a metastasis rate of 7.4%. PTC in the lower pole of thyroid and the number of cervical lymph nodes with metastasis > 6 are independent risks for LNSS metastasis.

Review of thyroid stimulating hormone receptor and thyroid carcinoma / 中华内分泌外科杂志

Thyroid carcinoma is the most common malignant tumor in the endocrine diseases,with a rising morbidity.As more investigations were made,thyroid stimulating hormone receptor is showed up,and it is believed some contacts are existed between thyroid stimulating hormone receptor and thyroid carcinoma.We believe that making sure of these contacts can help patients in diagnosis,treatment,and prognosis.

BRAF-Activated Long Noncoding RNA Modulates Papillary Thyroid Carcinoma Cell Proliferation through Regulating Thyroid Stimulating Hormone Receptor / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The importance of long noncoding RNAs (lncRNAs) in tumorigenesis has recently been demonstrated. However, the role of lncRNAs in development of thyroid cancer remains largely unknown. MATERIALS AND METHODS: Using quantitative reverse transcription polymerase chain reaction, expression of three lncRNAs, including BRAF-activated long noncoding RNA (BANCR), papillary thyroid cancer susceptibility candidate 3 (PTCSC3), and noncoding RNA associated with mitogen-activated protein kinase pathway and growth arrest (NAMA), was investigated in the current study. RESULTS: Of the three lncRNAs (BANCR, PTCSC3, and NAMA), expression of BANCR was significantly up-regulated while PTCSC3 and NAMA were significantly down-regulated in papillary thyroid carcinoma (PTC) compared to that in normal tissue. BANCR-knockdown in a PTC-derived cell line (IHH-4) resulted in significant suppression of thyroid stimulating hormone receptor (TSHR). BANCR-knockdown also led to inhibition of cell growth and cell cycle arrest at G0/G1 phase through down-regulation of cyclin D1. In addition, BANCR was enriched by polycomb enhancer of zeste homolog 2 (EZH2), and silencing BANCR led to decreased chromatin recruitment of EZH2, which resulted significantly reduced expression of TSHR. CONCLUSION: These findings indicate that BANCR may contribute to the tumorigenesis of PTC through regulation of cyclin D1 and TSHR.

Comparative study on minimally invasive video-assisted thyroidectomy and total endoscopic thyroidecto-my / 中华内分泌外科杂志

Objective To analyze the advantages and disadvantages of minimally invasive video -assisted thyroidectomy(MIVAT)and totally endoscopic thyroidectomy (TET).Methods The study included 70 patients undergoing MIVAT and TET at Qilu Hospital of Shandong University and Yantai Yuhuangding Hospital from Jan . 2012 to Dec.2012.46 patients undergoing MIVAT(MIVAT group)and 24 patients undergoing TET(TET group) were analyzed retrospectively .Data of gender,age,operative data,intraoperative and postoperative complications , length of stay , feelings of patients and cosmetic effect were collected and analyzed .MIVAT and TET were com-pared in terms of safety ,trauma and cosmetic effect .Results ①About safety:no permanent recurrent laryngeal nerve(RLN)palsy and parathyroid injury occurred in either group .One case in MIVAT group and no case in TET group was converted to open surgery (2.2%vs 0%,P>0.05) .②About trauma:the intraoperative blood loss and postoperative drainage amount in the MIVAT group were significantly less than those in the TET group ( P 0.05).③About cosmetic effect:the 2 groups had similar patient satisfaction index .Conclusions The study shows similar results in safety and trauma .The 2 approaches are both safe and valid .Patients using the two ap-proaches are equally satisfied with the cosmetic effect .

Detection of RASSF2 and sFRP1 promoter region methylation in sporadic colorectal cancer patients / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the methylation in promtor region of RASSF2 and sFRP1 in sporadic colorectal cancer patients in order to provide screening method for early colorectal cancer.</p><p><b>METHODS</b>The methylation in promoter region of RASSF2 and sFRP1 in serum samples of 59 sporadic colorectal cancer patients and 59 healthy volunteers was detected by methylation specific PCR. The association between clinicopathological features of sporadic colorectal cancer patients and methylation in promoter region of RASSF2 and sFRP1 was analyzed.</p><p><b>RESULTS</b>The methylation rates of RASSF2 and sFRP1 gene in serum of 59 sporadic colorectal cancer patients were 27.1% and 30.5%, significantly higher than those in healthy volunteers(0%, both P<0.01). The methylation of RASSF2 or sFRP1 occurred in 29(49.2%) patients, which was significantly higer than the methylation rate of single gene(P<0.05). No association was found between methylation ratio of RASSF2 and sFRP1 and clinicopathological features in sporadic colorectal cancer patients.</p><p><b>CONCLUSIONS</b>Methylation in promoter region of RASSF2 and sFRP1 is often detected in serum of colorectal cancer patients. The combination detection of methylation for the two genes may provide information for early screening of colorectal cancer.</p>

Analysis on the Rate Estimation of Coal Pneumoconiosis Liability Insurance / 中国卫生经济

Objective:To assess the risk of coal pneumoconiosis and measure the gross premium rate of coal pneumoconiosis liability insurance. Methods: Based on the data related to pneumoconiosis risk from published materials in different provinces, it used pure premium method to measure the gross premium rate and made province-specific rate tables of this liability insurance according to risk classification. Results: Rate tables of Chongqing, Shaanxi, Shanxi, Shandong, Henan and Anhui were made according to type of work and form of coal’s ownership. Conclusion: The risks of coal pneumoconiosis among different provinces are very different. It’s necessary to make province-specific rate tables when developing business of coal pneumoconiosis liability insurance for different provinces.

Intraoperative neuromonitoring in thyroid gland surgery / 中华内分泌外科杂志

Objective To summarize the experience of intraoperative neuromonitoring (IONM) system for monitoring and protection of recurrent laryngeal nerve (RLN) during thyroid surgery.Methods A total of 220 cases were enrolled in this study (53 males and 167 females),with the median age of 38.2 years old.There were 85 cases of thyroid cancer,19 cases of thyroid benign tumor,90 cases of thyroid goiter,3 cases of Hashimoto's diseases,and 23 cases of hyperthyroidism.113 cases had tumors larger than 5 cm in diameter.During the procedure,2 recording needle electrodes were put in cricothyroid muscle and 1 stimulator electrode was explored in tracheoasophageal groove.If any RLN was fight there or nearby,doctors can see the electromyogram and hear the toothonk.With careful dissection,RLN can be found out till exploring into the larynx site.Results Electromyogram showed in 207 cases (278 nerves),and it didn't show in 13 cases,among whom 9 cases were false-negative because of system or annesthesia problems,4 cases didn't manage to have needle electrodes put in properly due to cricothyroid muscle being invaded.No permanent RLN paralysis occoured.Transient nerve paralysis occurred in 2 cases,who recovered in 1 month after operation.Conclusion IONM system is an effective way to avoid damage to RLN in thyroidectomy.

Fast track surgery in laparoscopy-assisted radical distal gastrectomy / 中华普通外科杂志

ObjectiveTo evaluate the safety and effectiveness of fast track surgery (FTS) in l aparoscopy-assisted radical distal gastrectomy (LADG) for gastric cancer.MethodsSixty-one patientswith distal gastric cancer were randomly divided into three groups:FTS + LADG group (n =19) undergoing LADG and FTS treatments,LADG group (n =22) undergoing LADG and traditional perioperative cares,and FTS + ODG ( open distal gastrectomy) group ( n =21 ) undergoing ODG and FTS treatments.FTS treatments included avoidance of mechanical bowel cleansing,restrictive perioperative intravenous infusion,early ambulation,early enteral nutrition.The age,sex,body weight,anastomotic mode,number of lymph node dissected,and tumor stage,serum albumin (ALB),blood urea nitrogen (BUN),C-reaction protein (CRP),flatus time,postoperative hospital stay,medical cost,and postoperative complications were compared between three groups. ResultsThe level of ALB in FTS + LADG group were higher than in LADG group at the 4th and 7th day after surgery ( P ＜ 0.05,P ＜ 0.01 ).Compared to LADG group,the variation of ALB from preoperation to 4th day after surgery in FTS + LADG group and FTS + ODG group was significant( P ＜ 0.01,P ＜ 0.05 ).CRP level between FTS + LADG group and FTS + ODG group were different significantly at 4th and 7th day after surgery ( P ＜ 0.05,P ＜ 0.05).FTS + LADG group has earlier recovery of gastrointestinal peristalsis than other two groups ( P ＜ 0.05,P ＜ 0.05 ).The medical cost in FTS + LADG group was less than in LADG group ( P =0.003 ),but higher than in FTS + ODG group (P ＜0.01 ).ConclusionsThe practice of FTS in LADG was safe,effective,improves nutritional status,eases stress reaction,accelerates gastrointestinal peristalsis and postoperative rehabilitation.

Minimally incisional video-assisted thyroidectomy on thyroid tumor / 中华内分泌外科杂志

Objective To explore the efficiency of transcervical video-assisted mini-incision thyroidectomy for thyroid tumor. Methods 53 patients with thyroid tumors who underwent the video-assisted mini-incision thyroidectomy from Jan. 2006 to Dec. 2009 in the Department of Thyroid Surgery, Yantai Yuhuangding Hospital,were retrospectively studied. Results All of the patients, except 5 cases converting to open thyroidectomy, underwent the video-assisted mini-incision thyroidectomy successfully. The temporary recurrent laryngeal nerve injury occurred in 1 patients. The subcutaneous fluid accumulation occurred in 3 patients. The skin burns around the incisions occurred in 3 patients. There was no complication of hematoma, hypocalcemia, or superior laryngeal nerve injuries. The follow-up period was from 3 months to 35 months. There was no evidence of recurrence and metastasis by postoperative ultrasonic and CT scan and radioiodine scintigraphy. Conclusions The video-assisted mini-incision thyroidectomy is feasible, safe, and with the advantage of cosmetic results. For thyroid tumor,especially thyroid cancer, longer follow-up is necessary to draw definitive conclusion on recurrence and survival rate.

Expression of T cell-specific transcriptional factor Elf-1 in mononu-clear cells, CD4~+ T cells and CD8~+ T cells from umbilical cord blood / 中国病理生理杂志

AIM: To investigate the expression level of transcriptional factor Elf-1 in mononuclear cells, CD4~+ and CD8~+ T cells from umbilical cord blood (UCB). METHODS: Real-time PCR with SYBR green I technique was used for detecting the Elf-1 expression in mononuclear cells, sorted CD4~+ and CD8~+ T cells from 12 cases of umbilical cord blood. The relative mRNA expression level of Elf-1 was analyzed by a formula of 2~(-△Ct)×100%. The expression level of β_2-microglobulin gene (β_2M) was used as an endogenous reference. The peripheral blood from 10 cases of healthy adults was served as control. RESULTS: Elf-1 mRNA expressed in all blood samples collected from both UCB and healthy adults. The expression level showed apparent diversity in different individuals. The relative mRNA expression of Elf-1 in both mononuclear cells (18.55%±2.48%) and CD8~+ T cells (3.52%±0.45%) from UCB were significantly higher than those from healthy adults (9.16%±1.92%, 2.02%±0.27%, respectively, P<0.01, P<0.05). CONCLUSION: The results of Elf-1 expression level from umbilical cord blood indicate that the over-expression of Elf-1 gene in mononuclear cells and CD8~+ T cells might be one of the features of T cell immune state in umbilical cord blood.

Minimally invasive video-assisted thyroidectomy on thyroid microcarcinoma / 中华内分泌外科杂志

Objective To explore the feasibility and efficiency of minimally invasive video-assisted thy-roidectomy on thyroid microcarcinoma, and sum up the experiences. Methods 9 patients with thyroid microcarci-noma who underwent the minimally invasive video-assisted thyroidectomy from Jan 2006 to Feb 2009, were retro-spectively studied. Results All of the patients, except 1 case converting to open thyroidectomy, underwent the minimally invasive video-assisted thyroidectomy successfully. The subcutaneous hydrops occurred in 1 patient. The skin burns around the incisions occurred in 1 patient. There were no complications of recurrent laryngeal nerve injuries, neck hematoma, hypocalcemia, or superior laryngeal nerve injuries. The follow-up period of 15 cases were from 3 months to 37 months. There were no evidences of recurrence and metastasis by both postopera-five ultrasonic/CT examination and radioiodine scintigraphy. Conclusions The minimally invasive video-assisted thyroidectomy is safe and feasible (similar to the traditional thyroidectomy), of gratifying cosmetic results, for low-risk differentiated thyroid microcarcinoma without lymph node metastasis. For the median-risk or high-risk, more patients, comparative studies and long follow-ups are necessary to draw definitive conclusions in terms of its recurrence and survival rate.

Clinical analysis of metastatic carcinoma in thyroid / 中华内分泌外科杂志

Objective To explore the clinical manifestations, diagnosis, treatment and prognosis of secondary thyroid neoplasms. Methods To retrospectively analyse clinical manifestations,examinations and treatment of 83 cases of secondary thyroid cancer, which were retrieved from CNKI (1994-2008). Results In all cases, 39 were male, 39 were female and 5 were unknow, aged from 16 to 78 years old, median age was 52. The primary cancers included 31 esophageal carcinomas, 12 lung cancers, 8 laryngopharynx cancers, 7 clear cell renal carcinomas, 6 rectal carcinomas, 6 melanomas, 3 breast cancers, 3 rhabdomyosarcoma, 2 gatric cancers, 1 liver cancer, 1 cervical cancer, 1 nonchromaffin paraganglioma,1 ovary cancer, 1 lip squamous cell carcinomac. 64 patients had undergone chemotherapy or surgery or radiotherapy. The intervals between finding of primary carcinoma and secondary thyroid carcinoma ranged from 1 month to 26 years with a median time of 2.5 years. Patients died of widespread metastases from 1 month to 12 years following the diagnosis of secondary carcinoma in thyroid, with a median time of 32 months. Conclusions Metastasis in the thyroid gland is uncommon and may cause a diagnostic problem. Metastatic cancer to the thyroid should be considered when a patient with histoy of malignant tumor has a recent found thyroid lesions. With the combination of Surgery, chemotherapy and radiotherapy, the survival of patients can be significantly prolonged.

Expression of T cell-specific transcriptional factor Elf-1 in mononu-clear cells,CD4~+ T cells and CD8~+ T cells from umbilical cord blood / 中国病理生理杂志

AIM:To investigate the expression level of transcriptional factor Elf-1 in mononuclear cells,CD4+ and CD8+ T cells from umbilical cord blood (UCB). METHODS: Real-time PCR with SYBR green I technique was used for detecting the Elf-1 expression in mononuclear cells,sorted CD4+ and CD8+ T cells from 12 cases of umbilical cord blood. The relative mRNA expression level of Elf-1 was analyzed by a formula of 2-△Ct?100%. The expression level of ?2-microglobulin gene (?2M) was used as an endogenous reference. The peripheral blood from 10 cases of healthy adults was served as control. RESULTS: Elf-1 mRNA expressed in all blood samples collected from both UCB and healthy adults. The expression level showed apparent diversity in different individuals. The relative mRNA expression of Elf-1 in both mononuclear cells (18.55%?2.48%) and CD8+T cells (3.52%?0.45%) from UCB were significantly higher than those from healthy adults (9.16%?1.92%,2.02%?0.27%,respectively,P