Background and aims:Inflammatory factors play significant roles in the development and occurrence of hepatocellular carcinoma(HCC).However,the tumor-protective functions of growth differentiation factors(GDFs)in HCC are yet to be clarified.In this study,we aimed to evaluate the expression levels of 10 GDFs in tumor and paratumor tissues from patients with HCC and perform in vitro and in vivo ex-periments to elucidate the role of GDF7 in regulating the proliferation and metastasis of HCC.Methods:The gene expression of 10 GDFs was compared between HCC and paratumors using The Cancer Genome Atlas dataset and patient-derived tissues.A tumor microarray containing 108 HCC tissue samples was used to explore the prognostic value of GDF7 expression.Loss-of-function experiments were also performed in vitro and in vivo to investigate the role of GDF7 in HCC.Results:The mRNA and protein levels of GDF7 were significantly lower in HCC tumors than in para-tumors(P＜0.001).Kaplan-Meier analysis showed that decreased GDF7 expression in HCC was asso-ciated with worse overall survival(5-year rate:61.8％vs.27.5％,P＜0.001)and increased recurrence risk(P＜0.001).Multivariate Cox regression analysis demonstrated that low GDF7 expression,the presence of microvascular invasion,and elevated alpha-fetoprotein(AFP)levels were independent risk factors for tumor recurrence and poor survival.Downregulation of GDF7 also increased the tumor growth in HCC cells and in an HCC xenograft model.GDF7 knockdown promoted migration and invasion via epithelial-mesenchymal transition.Meanwhile,a negative correlation between JunB proto-oncogene(JUNB)and GDF7 was observed in HCC tissues.Modulating JUNB levels altered GDF7 protein expression.Conclusions:GDF7 is a potential biomarker for predicting superior outcomes in patients with HCC.GDF7 amplification is a potential therapeutic option for HCC.

Objective:To evaluate the clinical value of blastocysts derived from poor-quality embryos after day 3 (D3) transplantation and frozen, to investigate the best embryo strategies of frozen-thawed blastocysts.Methods:A retrospective cohort study was conducted on 362 frozen-thawed blastocyst transfer cycles were retrospectively analyzed in the Department of Reproductive Medicine, Tianjin First Central Hospital from July 2015 to March 2020. According to the days of culture, they were divided into day 5 (D5) and day 6 (D6) blastocyst transplantation group; according to the number of embryos transferred, they were divided into single blastocyst transplantation group and double blastocyst transplantation group; according to the embryo quality, single blastocyst transplantation group was divided into single high-quality embryo group and single poor-quality embryo group. The double blastocyst transplantation group was divided into double high-quality blastocyst group, a high-quality embryo matching a poor-quality blastocyst transplantation group and the double poor-quality blastocyst group. Clinical pregnancy rate, implantation rate and multiple pregnancy rate were compared in each group.Results:The clinical pregnancy rate and the implantation rate of D5 blastocyst transplantation group in thawing cycles were higher than those of D6 blastocyst transplantation group [62.96% (102/162) vs. 42.50%(85/200), 53.98% (122/226) vs. 35.86% (104/290)](all P<0.001), and there was no statistically significant difference of early abortion rate and multiple pregnancy rate between the D5 and D6 groups ( P>0.05). There was no statistically significant difference in the clinical pregnancy rate and the early abortion rate of D5 single blastocyst groups and double blastocyst pregnancy ( P>0.05), the implantation rate of single blastocyst group was higher than that of the double blastocyst group [62.24% (61/98) vs. 47.66% (61/128)] while the multiple pregnancy rate of single blastocyst group was lower than that of the double blastocyst group [3.28% (2/61) vs. 46.34% (19/41)] ( P=0.029, P<0.001). The clinical pregnancy rate and the multiple pregnancy rate of D6 single blastocyst group were lower than those of D6 double blastocyst pregnancy group [35.45% (39/110) vs. 51.11% (46/90), 5.13% (2/39) vs. 26.09% (12/46)] ( P=0.026, P=0.009), there were no statistically significant differences in implantation rate and early abortion rate (all P>0.05). There were no statistically significant differences in clinical pregnancy rate, implantation rate, early abortion rate and multiple pregnancy rate between single high-quality embryo and poor-quality embryo group in D5 and D6 single blastocyst transplantation group (all P>0.05). There were no statistically significant differences in clinical pregnancy rate, implantation rate, early abortion rate, and multiple pregnancy rate of the double high-quality blastocyst transplantation group, a high-quality embryo matching a poor-quality blastocyst transplantation group and the double poor-quality blastocyst group in D5 ( P>0.05). The implantation rate of D6 double poor-quality embryo group [26.67% (24/90)] was lower than that of the double high-quality embryo group [47.62% (20/42)] and a high-quality embryo matching a poor-quality embryo group [43.75% (7/16)]( P=0.029), while there were no statistically significant differences in clinical pregnancy rate, early abortion rate, and multiple pregnancy rate (all P>0.05). Conclusion:After the high-quality D3 embryo transfer and freezing, the remaining poor-quality embryos can be cultured to form blastocyst, better clinical pregnancy rate was obtained for transfer the frozen-thawed blastocysts, it increases the value of embryos and reduces the cost of treatment. The quality of D5 blastocyst is better than that of D6 blastocyst. The single blastocyst transplantation of D5 is preferred to reduce the rate of multiple pregnancy without affecting the clinical pregnancy rate.

Objective:To find a suitable semen treatment method in in vitro fertilization (IVF). Methods:A prospective randomized double-blind design was performed, 300 infertile couples with female factors and no abnormal fertility test were selected for IVF-embryo transfer (ET), the patients were randomly divided into float group (FY group, n=100), density gradient group (DG group, n=100) and upstream group (SU group, n=100). The remaining sperm after fertilization were collected from three groups, and sections of electron microscope were prepared. The ultrastructure of sperm was observed by transmission electron microscope. The differences among the three groups of selected sperm DNA fragment index (DFI) and fertilization rate, cleavage rate, high-quality embryo rate and pregnancy rate during IVF-ET were compared. Results:The sperm DFI of the three groups after treatment was statistically different ( P=0.02), and the sperm DFI of the FY group was significantly lower than that of DG group and SU group (3.22%±2.73% vs. 8.31%±2.14% vs. 6.43%±2.56%). The rates of sperm head and tail plasma membrane integrity rate in FY group were significantly higher than those in the other two groups (92.0%±24.2% vs. 80.2%±29.5% vs. 73.2%±30.1% and 93.9%±1.2% vs. 80.1%±1.1% vs. 74.9%±1.2%), and the differences were statistically significant ( P=0.01, P=0.03). The high-quality embryo rate and the blastocyst formation rate of FY group were significantly higher than those of DG group and SU group [44.14% (452/1024) vs. 32.30% (292/904) vs. 32.46% (296/912) and 54.40% (396/728) vs. 43.52% (302/694) vs. 46.34% (330/712)], and the differences were statistically significant (all P<0.001). The pregnancy rate and the implantation rate in the fresh cycle FY group were significantly higher than those in DG and SU groups [57.14% (24/42) vs. 33.33% (16/48) vs. 35.56% (16/45) and 50.00% (30/60) vs. 27.45% (14/51) vs. 28.26% (13/46)], and the differences were statistically significant ( P=0.04, P=0.02). The pregnancy rate and implantation rate in the resuscitation cycle FY group were significantly higher than those in the DG and SU groups [52.38% (22/42) vs. 31.25% (10/32) vs. 37.14% (13/35) and 52.38% (22/42) vs. 29.41% (10/34) vs. 30.56% (11/36)], and the difference was statistically significant ( P=0.03, P=0.02). Conclusion:The Plankton method can reduce physical damage to sperm, improve the integrity rate of sperm plasma membrane and decrease sperm DFI, improve assisted reproductive technology outcome.

Objective:To evaluate the clinical value of blastocysts derived from poor-quality embryos after day 3 (D3) transplantation and frozen, to investigate the best embryo strategies of frozen-thawed blastocysts.Methods:A retrospective cohort study was conducted on 362 frozen-thawed blastocyst transfer cycles were retrospectively analyzed in the Department of Reproductive Medicine, Tianjin First Central Hospital from July 2015 to March 2020. According to the days of culture, they were divided into day 5 (D5) and day 6 (D6) blastocyst transplantation group; according to the number of embryos transferred, they were divided into single blastocyst transplantation group and double blastocyst transplantation group; according to the embryo quality, single blastocyst transplantation group was divided into single high-quality embryo group and single poor-quality embryo group. The double blastocyst transplantation group was divided into double high-quality blastocyst group, a high-quality embryo matching a poor-quality blastocyst transplantation group and the double poor-quality blastocyst group. Clinical pregnancy rate, implantation rate and multiple pregnancy rate were compared in each group.Results:The clinical pregnancy rate and the implantation rate of D5 blastocyst transplantation group in thawing cycles were higher than those of D6 blastocyst transplantation group [62.96% (102/162) vs. 42.50%(85/200), 53.98% (122/226) vs. 35.86% (104/290)](all P<0.001), and there was no statistically significant difference of early abortion rate and multiple pregnancy rate between the D5 and D6 groups ( P>0.05). There was no statistically significant difference in the clinical pregnancy rate and the early abortion rate of D5 single blastocyst groups and double blastocyst pregnancy ( P>0.05), the implantation rate of single blastocyst group was higher than that of the double blastocyst group [62.24% (61/98) vs. 47.66% (61/128)] while the multiple pregnancy rate of single blastocyst group was lower than that of the double blastocyst group [3.28% (2/61) vs. 46.34% (19/41)] ( P=0.029, P<0.001). The clinical pregnancy rate and the multiple pregnancy rate of D6 single blastocyst group were lower than those of D6 double blastocyst pregnancy group [35.45% (39/110) vs. 51.11% (46/90), 5.13% (2/39) vs. 26.09% (12/46)] ( P=0.026, P=0.009), there were no statistically significant differences in implantation rate and early abortion rate (all P>0.05). There were no statistically significant differences in clinical pregnancy rate, implantation rate, early abortion rate and multiple pregnancy rate between single high-quality embryo and poor-quality embryo group in D5 and D6 single blastocyst transplantation group (all P>0.05). There were no statistically significant differences in clinical pregnancy rate, implantation rate, early abortion rate, and multiple pregnancy rate of the double high-quality blastocyst transplantation group, a high-quality embryo matching a poor-quality blastocyst transplantation group and the double poor-quality blastocyst group in D5 ( P>0.05). The implantation rate of D6 double poor-quality embryo group [26.67% (24/90)] was lower than that of the double high-quality embryo group [47.62% (20/42)] and a high-quality embryo matching a poor-quality embryo group [43.75% (7/16)]( P=0.029), while there were no statistically significant differences in clinical pregnancy rate, early abortion rate, and multiple pregnancy rate (all P>0.05). Conclusion:After the high-quality D3 embryo transfer and freezing, the remaining poor-quality embryos can be cultured to form blastocyst, better clinical pregnancy rate was obtained for transfer the frozen-thawed blastocysts, it increases the value of embryos and reduces the cost of treatment. The quality of D5 blastocyst is better than that of D6 blastocyst. The single blastocyst transplantation of D5 is preferred to reduce the rate of multiple pregnancy without affecting the clinical pregnancy rate.

Objective:To find a suitable semen treatment method in in vitro fertilization (IVF). Methods:A prospective randomized double-blind design was performed, 300 infertile couples with female factors and no abnormal fertility test were selected for IVF-embryo transfer (ET), the patients were randomly divided into float group (FY group, n=100), density gradient group (DG group, n=100) and upstream group (SU group, n=100). The remaining sperm after fertilization were collected from three groups, and sections of electron microscope were prepared. The ultrastructure of sperm was observed by transmission electron microscope. The differences among the three groups of selected sperm DNA fragment index (DFI) and fertilization rate, cleavage rate, high-quality embryo rate and pregnancy rate during IVF-ET were compared. Results:The sperm DFI of the three groups after treatment was statistically different ( P=0.02), and the sperm DFI of the FY group was significantly lower than that of DG group and SU group (3.22%±2.73% vs. 8.31%±2.14% vs. 6.43%±2.56%). The rates of sperm head and tail plasma membrane integrity rate in FY group were significantly higher than those in the other two groups (92.0%±24.2% vs. 80.2%±29.5% vs. 73.2%±30.1% and 93.9%±1.2% vs. 80.1%±1.1% vs. 74.9%±1.2%), and the differences were statistically significant ( P=0.01, P=0.03). The high-quality embryo rate and the blastocyst formation rate of FY group were significantly higher than those of DG group and SU group [44.14% (452/1024) vs. 32.30% (292/904) vs. 32.46% (296/912) and 54.40% (396/728) vs. 43.52% (302/694) vs. 46.34% (330/712)], and the differences were statistically significant (all P<0.001). The pregnancy rate and the implantation rate in the fresh cycle FY group were significantly higher than those in DG and SU groups [57.14% (24/42) vs. 33.33% (16/48) vs. 35.56% (16/45) and 50.00% (30/60) vs. 27.45% (14/51) vs. 28.26% (13/46)], and the differences were statistically significant ( P=0.04, P=0.02). The pregnancy rate and implantation rate in the resuscitation cycle FY group were significantly higher than those in the DG and SU groups [52.38% (22/42) vs. 31.25% (10/32) vs. 37.14% (13/35) and 52.38% (22/42) vs. 29.41% (10/34) vs. 30.56% (11/36)], and the difference was statistically significant ( P=0.03, P=0.02). Conclusion:The Plankton method can reduce physical damage to sperm, improve the integrity rate of sperm plasma membrane and decrease sperm DFI, improve assisted reproductive technology outcome.

Objective To evaluate the prognostic significance of the candidate selection Hangzhou criteria for liver transplantation of HCC patients undergoing hepatectomy.Methods 199 HCC patients undergoing hepatectomy between 2009 and 2011 were enrolled retrospectively.Predictors of survival were identified using the Kaplan-Meier method.The disease state was staged by the Hangzhou criteria (HC) and Milan staging systems.Calculating the area under the receiver operating characteristic (ROC) curve (AUC) evaluates the discriminatory ability for the prediction of survival of both staging system.Results Portal vein thrombosis,poor differentiation,and tumor size (＞ 8 cm) were independent risk factors for survival after hepatectomy.Milan criteria and Hangzhou criteria functioned well in predicting tumor-recurrence.For 1-year AUROC,the AUROC for Milan criteria and Hangzhou criteria are 0.602 and 0.741,respectively.For 3-year AUROC,the AUROC for Milan criteria and Hangzhou criteria are 0.643 and 0.733,respectively.Conclusions The HC were shown to be a promising survival predictor in a Chinese cohort of HCC patients after hepatectomy.

OBJECTIVE To analyze trimethylation of genome-wide histone H3 lysine 4(H3K4met3) induced by silicon dioxide(SiO2)through chromatin immunoprecipitation linked to microarrays(ChIP-chip)in lung fibroblast(LF)of rats. METHODS A primary co-culture model of rat alveolar macrophages (AM)and LF in vitro. AM were exposed to 100 mg · L-1 free SiO2 for 24 h,before LF were collected and the phenotype of LF was determined after transdifferentiation by immunohistochemistry. ChIP-chip was used to profile the variations of trimethylation in H3K4 of lung fibroblasts in CpG island regions. ChIP-qPCR was used to validate the microarray results. The mRNA expression of nfib and kpna3 was analyzed by qRT-PCR. RESULTS Totally 1815 (518 increased and 1297 decreased) genes of H3K4met3 displayed significant differences in SiO2 100 mg·L-1 group compared with control group(Cy3/Cy5 value>2.0 or <0.5,NimbleScan V2.5 software). The results of ChIP-qPCR were quite consistent with those of microarray. CONCLUSION There are significant differences in methylation of genome-wide H3K4 between SiO2 100 mg·L-1 group and control group. These novel candidate genes may become potential biomarkers or new interfered targets.

Objective To investigate the clinical values of next-generation sequencing (NGS) technology in diagnosis of miscarried chorionic villi genetic disorders. Methods Patients who underwent miscarriage (n=87) were enrolled in this study. Among all patients, 32 cases were of recurrent miscarrage and 55 cases were of sporadic miscarriage. In all collected patients, 35 women were 35 years or older while other 52 women were less than 35 years old. Positive detection rate and the abnormal detection rate were compared between these two methods. Chromosomes abnormal rates were also compared among different types of miscarrage and different ages. All aborted villi tissue were analyzed by NGS of whole genome and G-band?ing karyotype. Results The successful detection rate of chorionic villi by NGS (100.00%) was higher than that of G-band?ing karyotype (74.71%), and the detection rate of abnormal chorionic villi by NGS (58.62%) was also higher than that of G-banding karyotype (50.77%). Three cases of chromosome structure anomaly were found in those 51 chromosome anomalies (5.88%). Other 48 cases of chromosome anomalies were aneuploidy anomalies (94.12%) include 39 cases of trisomy, 2 cases of double trisomy and 1 case of triple trisomy and 6 cases of monomer. On the other hand, 32 cases of chromosome aneuploi?dy anomalies were found in 33 chromosome anomalies by G-banding karyotype, which include 24 cases of trisomy, 2 cases of double trisomy, 1 case of triple trisomy, 5 cases of monomer and 1 case of chromosome structure anomaly. Most NGS re?sults (n=64) were in agreement with G-banding karyotype but with 1 case of discrepancy. Chromosomal abnormality rate de?tected by NGS in sporadic miscarrage group and recurrent spontaneous miscarrage group were 60.00%and 56.25%respective?ly. There was no significant difference (P>0.05). Chromosomal abnormality rate picked by NGS in women aged≥35 years old (71.43%) was higher than that in women<35 years old (50.00%) with statistically significant difference (P<0.05). Conclu?sion NGS technology showed highly accuracy in detecting chromosomal abnomality from villi tissue. Therefore, it could help to detect genetic disorders of miscarrage. It is useful to determine the reasons of miscarrage and guide the next pregnancy.

Objective:To explore the feasibility of the newly developed micro‐irrigation tube in the application to fistula complicated with empyema after esophagectomy .Methods:From Jan 2012 to Dec 2014 ,5 patients with fistula complicated with empyema after esophagectomy ,admitted in Zhongshan Hospital ,Fudan University ,accepted the therapy with a new type of micro‐irrigation tube .Among them ,4 patients were male and the other 1 was female .Three patients accepted left transthoracic esophagectomy and the other 2 underwent right thoracoabdominal esophagectom .Four patients got anastomotic stoma fistula , and the other 1 patient got stump fistula .The micro‐irrigation tube was composed of anesthesia epidural catheter ,double pipe joint and bottle water seal joint .Along the chest dralnage tube ,the micro‐irrigation tube was placed into the empyema cavity . Results:There was no serious adverse reaction except for the symptoms of cough and acid reflux in 2 patients .All the 5 patients were discharged smoothly with fistula healed and chest tube drawn .Conclusions:The application of micro‐irrigation tube in fistula complicated with empyema after esophagectomy is safe and feasible .

Objective To investigate appropriate diagnosis and treatment of solitary fibrous tumor of the pleura (SFTP).Methods Clinical and pathological data of ten patients treated in our hospital from 2002 to 2007 were reviewed. Results Our series consisted of three men and seven women. In two patients correct diagnosis was made before operation through ultrasonography-gnided core needle biopsy. All the patients were treated surgically including three resected by video-assisted thoracic surgery (VATS). Histopathologically, five tumors were malignant and the other five were benign. Immunohistochemical staining showed malignant SFTP (3/5) were less frequently positive for CD34 than benign group (5/5). Nestin was only detected in malignancies (2/5), which were negative for CD34. Except for one, all patients were followed-up for 6 to 35 months (mean 17.3 months). One patient experienced a recurrence and one died of brain metastasis. Conclusion Ultrasonography-guided core needle biopsy combined with immunohistochemical analysis is a safe and rapid method to provide a confirmatory diagnosis before surgery. For smaller, pedunculated tumors, VATS may be a bettor approach. Besides, we speculated CD34-negative and nestin-posifive might be a malignant marker for SFTP.