Objective:To detect the expression of long non-coding RNA (LncRNA) ARAP1-AS1 in pancreatic cancer, and to preliminarily explore its effects on the biological behaviors of proliferation, apoptosis, migration and invasion of pancreatic cancer cell.Methods:The pancreatic cancer tissue specimens and corresponding paracancerous tissue specimens of 25 patients were collected, and the expression of ARAP1-AS1 was detected by qPCR. Human pancreatic cancer cell line PANC-1 was cultured in vitro and divided into control group, siRNA-control group (transfected with siRNA control sequence), knockout group (transfected with ARAP1-AS1 siRNA), pcDNA3.1-control group (transfected with pcDNA3.1) and overexpression group (transfected with pcDNA3.1-ARAP1-AS1), qPCR method was used to detect the transfection efficiency, CCK-8 method was used to detect the cell proliferation ability, flow cytometry was used to detect the cell apoptosis, scratch test was used to detect the cell migration ability, Transwell method was used to detect the cell invasion ability, Western blot (WB) method was used to detect the expression of proliferating cell nuclear antigen (PCNA), B lymphoma-2 protein (Bcl-2), Bcl-2 related X protein (Bax), matrix metalloproteinase-9 (MMP-9) proteins.Results:The expression level of ARAP1-AS1 in pancreatic cancer tissues was significantly higher than that in adjacent tissues (2.26±0.13 vs 1.00±0.00) ( P<0.05). Compared with the siRNA-control group, the ARAP1-AS1 level (1.01±0.02 vs 0.29±0.03), PCNA, Bcl-2, MMP-9 protein levels, cell OD value (0.57±0.05 vs 0.23±0.03), scratch healing rate (78.53±7.02 vs 48.60±5.26), and number of invasions (229.63±22.59 vs 104.25±15.04) in PANC-1 cells of the knockout group were significantly reduced ( P<0.05), the Bax protein level and the apoptosis rate (4.52±0.42 vs 32.40±1.84) were significantly increased ( P<0.05). Compared with the pcDNA3.1-control group, the ARAP1-AS1 level (1.02±0.03 vs 2.06±0.08), PCNA, Bcl-2, MMP-9 protein levels, cell OD value (0.57±0.05 vs 0.90±0.08), scratch healing rate (77.65±6.67 vs 91.22±7.34), and number of invasions (225.34±19.65 vs 327.50±25.40) in PANC-1 cells of the overexpression group were significantly increased ( P<0.05), the Bax protein level and the apoptosis rate (4.58±0.48 vs 2.29±0.24) were significantly reduced ( P<0.05) . Conclusion:LncRNA ARAP1-AS1 is highly expressed in pancreatic cancer, which can promote the proliferation, migration and invasion of pancreatic cancer cells PANC-1, and reduce cell apoptosis.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with resistance to thyroid hormone syndrome (RTH). METHODS: Exons 7 to 10 of the THRbeta gene were sequenced for the proband and members of his pedigree. RESULTS: Three patients from the pedigree were identified. All have presented with palpitation, fatigue, goiter, elevated free thyroid hormone and free triiodothyronine, and normal or elevated thyrotropin. Genetic testing revealed that the proband, his mother, second sister and one of her daughters had carried a heterozygous c.1336T>A variant of the THRbeta gene, which resulted in substitution of Cysteine by Serine at position 446. The variant was unreported previously. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.1336T>A(p.Cys446Ser) variant of THRbeta gene was predicted to be lilely pathogenic(PM1+PM2+PM5+PP3). CONCLUSION The c.1336T>A variant, identified in the exon 10 of the THRbeta gene, probably underlay the RTH in this pedigree. Genetic testing has validated the clinical diagnosis for this pedigree.
Female ; Genomics ; Heterozygote ; Humans ; Mothers ; Mutation ; Pedigree

Objective:To compare long-term efficacy between watch and wait (W&W) strategy and total mesorectal excision (TME) in patients who were diagnosed with locally advanced rectal cancer (LARC) and attained clinical complete response (cCR) after neoadjuvant chemoradiotherapy (nCRT).Methods:A retrospective cohort study was carried out. A total of 238 patients with stage II-III LARC exhibiting cCR after nCRT in Sun Yat-sen University Cancer Center from September 16, 2010 to January 9, 2018 were enrolled. Patients who were diagnosed with other malignant tumor within 5 years, did not receive regular follow-up in our center for more than 1 year and had no complete examination items after nCRT were excluded. Of 238 patients, 151 were male and 87 were female with a median age of 57 (27-83) years old. According to TNM stage, 61 cases were cII, 177 cases were cIII. Concurrent chemoradiotherapy (CCRT) was performed in 20 patients. CCRT plus induction/consolidated chemotherapy was performed in 218 patients. Intensity-modulated radiotherapy (IMRT) was applied to radiotherapy. The median radiation dose was 50 Gy/25 Fr for both the primary tumor and clinical target volumes, and the total dose was 45.0 to 50.6 Gy for 227 patients. In 27 patients, single-agent fluorouracil or capecitabine was used as concurrent chemotherapy. But in the other 211 patients, a combined regimen of oxaliplatin and fluorouracil or capecitabine was used. After nCRT, 59 and 179 patients received W&W (W&W group) and TME 6-12 weeks later (TME group), respectively. After the ending of treatment, patient was interviewed one time every 3 months and after 3 years, one time every six months. Overall survival (OS) rate, distant-metastasis-free survival (DMFS) rate, and local-recurrence-free survival (LRFS) rate were compared between two groups. The salvage treatment and sphincter preservation rate were analyzed. The survival curve was drawn with Kaplan-Meier method and evaluated by log-rank method.Results:In the cases treated with TME, the median interval from nCRT to surgery was 59 days. The postoperative pCR rate was 63.1%(113/179). The median follow-up time of the whole cohort was 41.8 (12.0-99.0) months. The 3-year and 5-year OS rates were 98.4% and 96.5%; the 3-year and 5-year LRFS rates were 96.5% and 96.5%; the 3- and 5-year DMFS rates were 91.0% and 87.9%, respectively. The 3-year OS rates in the W&W group and the TME group were 100% and 97.9%; the 5-year OS rates in W&W group and the TME group were 90.6% and 97.9% ( P=0.339); The 3-year local recurrence rate (LRR) in the W&W group was 12.9% (7 cases recurred within 2 years), which was significanthy higher then that in the TME group (0.6%, P=0.003). Salvage surgery was successful in 5/6 cases. After salvage surgery, LRFS rate was not significantly different between the two groups ( P=0.137). The 3-year DMFS rate in the W&W group and the TME group were 88.4% and 81.1%, whose difference was not significant ( P=0.593). Recurrence with simultaneous metastasis was seen in 3/7 cases of the W&W group. The sphincter was preserved in 89.8% (53/59) of patients in the W&W group, which was significantly higher than 73.7% (132/179) in the TME group ( P<0.001). When distance of tumor from the anal verge was ≤ 5 cm, the sphincter preservation rate (SPR) in the W&W group was 88.0% (44/50), which was significantly higher than the 54.4% (56/103) in the TME group ( P<0.001). Conclusions:W&W is safe and feasible for patients with LARC and cCR after nCRT. The results should be verified by further clinical trials.

Objective:To compare long-term efficacy between watch and wait (W&W) strategy and total mesorectal excision (TME) in patients who were diagnosed with locally advanced rectal cancer (LARC) and attained clinical complete response (cCR) after neoadjuvant chemoradiotherapy (nCRT).Methods:A retrospective cohort study was carried out. A total of 238 patients with stage II-III LARC exhibiting cCR after nCRT in Sun Yat-sen University Cancer Center from September 16, 2010 to January 9, 2018 were enrolled. Patients who were diagnosed with other malignant tumor within 5 years, did not receive regular follow-up in our center for more than 1 year and had no complete examination items after nCRT were excluded. Of 238 patients, 151 were male and 87 were female with a median age of 57 (27-83) years old. According to TNM stage, 61 cases were cII, 177 cases were cIII. Concurrent chemoradiotherapy (CCRT) was performed in 20 patients. CCRT plus induction/consolidated chemotherapy was performed in 218 patients. Intensity-modulated radiotherapy (IMRT) was applied to radiotherapy. The median radiation dose was 50 Gy/25 Fr for both the primary tumor and clinical target volumes, and the total dose was 45.0 to 50.6 Gy for 227 patients. In 27 patients, single-agent fluorouracil or capecitabine was used as concurrent chemotherapy. But in the other 211 patients, a combined regimen of oxaliplatin and fluorouracil or capecitabine was used. After nCRT, 59 and 179 patients received W&W (W&W group) and TME 6-12 weeks later (TME group), respectively. After the ending of treatment, patient was interviewed one time every 3 months and after 3 years, one time every six months. Overall survival (OS) rate, distant-metastasis-free survival (DMFS) rate, and local-recurrence-free survival (LRFS) rate were compared between two groups. The salvage treatment and sphincter preservation rate were analyzed. The survival curve was drawn with Kaplan-Meier method and evaluated by log-rank method.Results:In the cases treated with TME, the median interval from nCRT to surgery was 59 days. The postoperative pCR rate was 63.1%(113/179). The median follow-up time of the whole cohort was 41.8 (12.0-99.0) months. The 3-year and 5-year OS rates were 98.4% and 96.5%; the 3-year and 5-year LRFS rates were 96.5% and 96.5%; the 3- and 5-year DMFS rates were 91.0% and 87.9%, respectively. The 3-year OS rates in the W&W group and the TME group were 100% and 97.9%; the 5-year OS rates in W&W group and the TME group were 90.6% and 97.9% ( P=0.339); The 3-year local recurrence rate (LRR) in the W&W group was 12.9% (7 cases recurred within 2 years), which was significanthy higher then that in the TME group (0.6%, P=0.003). Salvage surgery was successful in 5/6 cases. After salvage surgery, LRFS rate was not significantly different between the two groups ( P=0.137). The 3-year DMFS rate in the W&W group and the TME group were 88.4% and 81.1%, whose difference was not significant ( P=0.593). Recurrence with simultaneous metastasis was seen in 3/7 cases of the W&W group. The sphincter was preserved in 89.8% (53/59) of patients in the W&W group, which was significantly higher than 73.7% (132/179) in the TME group ( P<0.001). When distance of tumor from the anal verge was ≤ 5 cm, the sphincter preservation rate (SPR) in the W&W group was 88.0% (44/50), which was significantly higher than the 54.4% (56/103) in the TME group ( P<0.001). Conclusions:W&W is safe and feasible for patients with LARC and cCR after nCRT. The results should be verified by further clinical trials.

Objective To investigate the feasibility of non-operative management (NOM) by comparing the therapeutic effects between NOM and total mesorectal excision (TME) for rectal cancer patients with clinical complete response (cCR) after neo-adjuvant chemoradiotherapy.Methods A total of 135 patients with stage Ⅱ/Ⅲ rectal cancer who obtained cCR after neo-adjuvant chemoradiotherapy in Sun Yat-sen University Cancer Center from 2006 to 2016 were recruited and assigned into the NOM (n =43) and standard operative management (SOM) groups (n=92).The local recurrence rate,accumulative local control (LC) rate after salvage therapy,disease-free survival (DFS),overall survival (OS) and sphincter preservation rate were statistically compared between two groups.Kaplan-Meier analysis and log-rank test were utilized to calculate the LC,OS and DFS.Chi-square test was performed to calculate the sphincter preservation rate.Results The mean follow-up duration was 39 months (range:10-127 months).Of 135 patients,the local recurrence rate and distant metastasis rate were 3.7％ and 11.1％,and the 3-year DFS and OS were 90.5％ and 97.0％.In the NOM and SOM groups,the 3-year DFS were 87％ and 93％,and the 5-year DFS were 73％ and 87％(P=0.089).The 3-year OS were 98％ and 99％,and the 5-year OS were 98％ and 97％ (P=0.578).In the NOM group,the local recurrence rate was 12％ (n =5),80％ of patients received salvage treatment and the accumulative LC rate was calculated as 98％.In the SOM group,the local recurrence rate was 0,which was significantly lower than that in the NOM group (P=0.O10).In the NOM group,the sphincter preservation rate was 93％,significantly higher compared with 70％ in the SOM group (P=0.030).Conclusions It is feasible for rectal cancer patients with cCR to receive NOM following neo-adjuvant chemoradiotherapy.Partial locally recurrent patients can be healed by timely salvage therapy,thereby averting TME and relevant complications and enhancing the quality of life of rectal cancer patients.

Objective To investigate the efficacy and toxicities of neoadjuvant chemoradiotherapy (neoCRT) in the management of unresectable locally advanced adherent colon cancer (LAACC).Methods A retrospective analysis was performed on the clinical records of 40 patients with initially diagnosed unresectable LAACC who received preoperative neoCRT in our center from October 2010 to December 2015.Results Thirty-nine patients completed the preoperative neoCRT.Thirty-four patients underwent radical resection after neoCRT, and the R0 resection rate, pathological complete response rate (pCR), tumor downstaging rate, nodal downstaging rate, and clinical downstaging rate were 91%, 24%(8/34patients), 76%(26/34patients),100%(32/32patients), and 94%(32/34patients), respectively.Among the 21 patients with bladder invasion, the full bladder was preserved in 7 patients (33%) and partial cystectomy was performed in 11 patients (52%).During the course of neoCRT, the grade 3-4 hematologic toxicity rate, grade 3 hand-foot syndrome rate, grade 3 radiodermatitis, and incomplete intestinal obstruction rate were 23%, 3%, 3%, and 5%, respectively.The 3-year sample size was 25 patients.For all the patients, the 3-year overall survival (OS) and progression-free survival (PFS) rates were 75% and 80%, respectively.Of the 34 patients who received surgical radical resection, the 3-year OS and disease-free survival (DFS) rates were 87% and 81%, respectively.In addition, local tumor recurrence was identified in 3 patients, and distant metastasis was identified in 6 patients.Conclusions NeoCRT is an effective treatment for unresectable LAACC that results in significant tumor downstaging and enhanced R0 resection rate without an increase in surgical complications.The patients treated with radical surgical resection after neoCRT show a satisfactory short-term outcome.Further studies will be required to determine the clinical value of neoCRT in treating LAACC.

Objective To evaluate the mid-to long-term survival benefits of preoperative sandwich-like neoadjuvant chemoradiotherapy (CRT) in patients with locally advanced rectal cancer (LARC).Methods A total of 45 LARC patients who underwent neoadjuvant sandwich CRT in the form of XELOX regimen prior to,concurrently with,and following volumetric modulated arc radiotherapy (VMAT) in 2012 were enrolled in this study.VMAT was given at a gross tumor volume dose of 50 Gy in 25 fractions,and a clinical target volume dose of 45-46 Gy in 25 fractions.Total mesorectal excision was performed 6 to 8 weeks after completion of VMAT.The overall survival (OS) and disease-free survival (DFS) were determined by the Kaplan-Meier method,and survival comparison and univariate prognostic analysis were performed using the log-rank test.Results The median follow-up time was 46.7 months.There was no local recurrence detected among the patients.The 3-year distant metastasis (DM) rate was 18%,and the 3-year OS and DFS were 96% and 84%,respectively.Univariate analysis indicated that perineural invasion,N1-N2 pathology (pathological stage Ⅲ),and Ca-199>35 U/ml before treatment were risk factors for DM (P=0.000,0.000,and 0.013,respectively).Conclusions The significant short-term efficacy of preoperative sandwich-like neoadjuvant CRT can be extended to a positive mid-term survival in LARC patients.However,further phase Ⅲ clinical studies will be needed to confirm this finding.

Objective To investigate the influence of maternal high-protein diet during pregnancy and lactation on the growth of male rat offspring.Methods Female Wistar rats were mated with male rats and the pregnant ones were randomly assigned into three groups (n=10):Group Ⅰ,Group Ⅱ and Group Ⅲ.Three isocaloric purified diets,which were Diet Ⅰ (protein,14.0％;carbohydrate,69.3％;fat,16.7％),Diet Ⅱ (protein,24.0％;carbohydrate,59.3％;fat,16.7％) and Diet Ⅲ (protein,34.0％;carbohydrate,49.3％;fat,16.7％),were offered ad libitum to the rats in the three groups,respectively.On the 3rd day after birth,only three male rat offspring of each litter were kept.All male rat offspring at the age of 21 days were weaned onto the same normal diets until 77-day-old.Food intake (FI),body weight (BW),body length (BL) and visceral fat mass (VFM) of them were recorded.Blood lipids levels on the 21st,49th and 77th days after birth were detected.Weight gain (WG),food efficiency (FE) and percentage of VFM (VFM ％) were calculated to assess the growth of male rat offspring.One-way ANOVA,LSD or Tamhane T2 test was used for statistical analysis.Results (1) On the 3rd day after birth,male rat offspring in Group Ⅱ and Group Ⅲ had higher fast BW than those in Group Ⅰ [(9.77±0.81),(9.58±0.54) and (9.17±0.68) g;F=3.222,P=0.026].On the 7th,14th and 21st days after birth,male rat offspring in Group Ⅱ [(17.59± 1.24),(39.51 ±2.68) and (67.77±4.22) g] had higher BW than those in Group Ⅰ [(15.96±1.17),(35.35±4.11) and (63.43±3.98) g] and Group Ⅲ [(16.52±1.05),(37.06±3.27) and (64.43±3.81) g] (F=23.684,17.070 and 84.195,all P＜0.001).Male rat offspring's BL of Group Ⅱ were longer than those of Group Ⅰ on the 7th and 14th days after birth[(7.53±0.29) vs (7.53± 0.29) cm,(10.38 ±0.24) vs (9.99± 0.30) em,both P＜0.05].BL of Group Ⅲ was longer than that of Group Ⅰ,but shorter than that of Group Ⅲ on the 14th day after birth[(10.22 ± 0.25) vs (9.99± 0.30) cm,(10.22 ± 0.25) vs (10.38±0.24) cm,both P＜0.05].Male rat offspring's average WG from the 3rd to the 7th and the 7th to the 14th day after birth were higher in Group Ⅱ [(7.70±0.41) and (22.08±1.20) g] and Group Ⅲ [(7.00±0.40) and (20.75± 1.72) g] than in Group Ⅰ [(6.73±0.55) and (19.68± 1.73) g] (F=86.925 and 38.876,both P＜0.001),and the differences between Group Ⅱ and Group Ⅲ were statistically significant (both P＜0.05).(2) No significant differences in WG,FI and FE was observed among the three groups (all P＞0.05).Male rat offspring's BL on the 49th day after birth was longer in Group Ⅱ than in Group Ⅰ and Group Ⅲ [(22.03±0.26),(21.57±0.43) and (21.77±0.33) cm,F=3.222,P=0.026).VFM ％ of Group Ⅱ (3.87±0.32 and 5.13±0.32) and Group Ⅲ (3.90±0.27 and 5.15±0.33) on the 49th and 77th days after birth were higher than those of Group Ⅰ (3.50±0.34 and 4.68±0.38) (F=3.631 and 3.611,both P＜0.05).(3) Triglyceride (TG) level was higher in Group Ⅰ than in Group Ⅱ and Group Ⅲ on the 21st day after birth [(1.12±0.13),(0.89±0.10) and (0.97±0.12) mmol/L,F=7.283,P=0.004].However,Group Ⅱ and Group Ⅲ had a higher level of TG than Group Ⅰ on the 77th day after birth[(2.64±0.37),(2.43±0.32) and (1.90±0.21) mmol/L,F=12.321,P＜0.001].Conclusion Maternal high-protein diet can increase the birth weight of male rat offspring to a certain extent,which is influenced by carbohydrate content.Moreover,male rat offspring of dams fed with high-protein diet during pregnancy and lactation will have increased visceral fat accumulation and serum TG level during adulthood.

Objective To investigate the prognostic value of American Joint Committee on Cancer-tumor regression grading ( AJCC-TRG) combined with ypTN stage in patients with locally advanced rectal cancer (LARC),who were treated with neoadjuvant chemoradiotherapy,and to identify the subgroups with the worst prognosis. Methods A total of 263 patients with LARC,including 176 males and 87 females,with a median age of 55 years,were admitted to Sun Yat-sen University Cancer Center from 2004 to 2012.All the patients received neoadjuvant chemoradiotherapy before surgery and underwent total mesorectal excision at 6 to 8 weeks after radiotherapy. All the surgical specimens were reevaluated according to the AJCC ( 7th edition)-TRG system and ypTN staging criteria. The prognostic prediction by TRG combined with ypTN was evaluated using survival analysis. The Kaplan-Meier method was used to calculate the rates of overall survival ( OS ) , disease-free survival ( DFS ) , local recurrence-free survival ( LRFS ) , and distant metastasis-free survival ( DMFS ) . The log-rank test was used for survival comparison and univariate prognostic analysis. Results The median follow-up was 601 months. The 5-year rates of OS, DFS, LRFS, and DMFS for all patients were 800%,750%,970%,and 810%,respectively. There were significant differences in OS, DFS,and DMFS between different ypT/TRG subgroups and different ypN/TRG subgroups (all P<005). ypT3-4/TRG 2-3 and ypN1-2/TRG 2-3 subgroups showed the worst prognosis. The 5-year rates of OS,DFS, and DMFS of the two subgroups were 669%/560%, 522%/414%, and 609%/460%, respectively. Conclusions A combination of AJCC-TRG system and ypTN staging can better predict the prognosis of LARC and identify the subgroups with the worst prognosis, which may provide a clinical guidance for postoperative individualized decision on adjuvant therapy for LARC.

Objective To clone express and purify Schistosoma japonicum fructose?1 6?bisphosphate aldolase SjFBPA in E. coli and observe its expression in different developmental stages of S. japonicum. Methods FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid and inducibly expressed with IPTG in E. coli BL21. SDS?PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA rSjFBPA . Then rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS?PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore SjFBPA mRNA was ana?lyzed in different developmental stages of S. japonicum by RT?PCR. Results SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro?tein could specifically reactive to the anti?His?tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT?PCR showed that SjFBPA mRNA was expressed in cercaria schistosomulum adult worm and egg of S. japonicum. Conclusion SjFBPA is successfully recombined and expressed in a prokaryotic system and SjFBPA mRNA is expressed in cercaria schistosomulum adult worm and egg of S. japonicum.