Objective To explore the effects from the synergy of substrate stiffness and hypoxia on epithelial mesenchymal transition (EMT) of colon cancer cells SW480 by simulating the microenvironment of human colon cancer tissues. Methods Polyvinyl alcohol gels with different stiffness ( 4. 5, 20, 40 kPa) were prepared to simulate the stiffness of each part of colon cancer tissues. The morphological change of cells on substrate with different stiffness was detected under simulated hypoxia ( CoCl2 ) environment. The expression of hypoxia inducible factor (HIF-1α), and EMT markers E-cadherin, Vimentin, Snail 1 were detected by Western blot. The mRNA expression of E-cadherin, Vimentin, Snail 1, matrix metalloproteinase-2 ( MMP-2), and MMP-9 was detected by quantitative real-time PCR ( qRT-PCR). Results Under simulated hypoxia environment, with the increase of substrate stiffness, the SW480 cells spreading area increased, and transformed from round shape into irregular polygon. The EMT of SW480 could be enhanced through up-regulating expression of Vimentin, Snail 1, MMP-2, MMP-9, and down-regulating expression of E-cadherin. Conclusions This study is important for exploring the synergistic effect of substrate stiffness and hypoxia on the EMT of colon cancer cells as well as the molecular mechanism.

Via an insufficient coat protein complex I (COPI) retrieval signal, the majority of SARS-CoV-2 spike (S) is resident in host early secretory organelles and a tiny amount is leaked out in cell surface. Only surface-exposed S can be recognized by B cell receptor (BCR) or anti-S therapeutic monoclonal antibodies (mAbs) that is the trigger step for B cell activation after S mRNA vaccination or infected cell clearance by S mAbs. Now, a drug strategy to promote S host surface exposure is absent. Here, we first combined structural and biochemical analysis to characterize S COPI sorting signals. A potent S COPI sorting inhibitor was then invented, evidently capable of promoting S surface exposure and facilitating infected cell clearance by S antibody-dependent cellular cytotoxicity (ADCC). Importantly, with the inhibitor as a probe, we revealed Omicron BA.1 S is less cell surface exposed than prototypes because of a constellation of S folding mutations, possibly corresponding to its ER chaperone association. Our findings not only suggest COPI is a druggable target against COVID-19, but also highlight SARS-CoV-2 evolution mechanism driven by S folding and trafficking mutations.

Objective:To summarize the experience and technical difficulties of complete thoracoscopic lobectomy(segmental) in the treatment of congenital pulmonary airway malformation in children.Methods:The clinical data, surgical videos and follow-up results of 38 children suffering from congenital pulmonary airway malformation and underwent complete thoracoscopic lobectomy in Children′s Hospital of Fudan University Xiamen Branch from January 2017 to December 2019 were retrospectively analyzed.A statistical comparisons of operative time, intraoperative blood loss, postoperative pain (the first time of getting out of bed), incision length and postoperative pulmonary complications were made between 28 children who underwent the total thoracoscopic lobectomy and 21 children who accepted open surgical lobectomy before January 2017 in Children′s Hospital of Fudan University Xiamen Branch.Results:Thirty-eight children with congenital pulmonary airway malformation were successfully operated by complete thoracoscopy.There were 28 lobectomies, 6 segmental lobectomies and 4 wedge lobectomies.Postoperative follow-up was conducted for more than 3 months, and no serious surgical complications occurred.Chest radiograph of 2 children with the right lower lobectomy at 48 hours after surgery had the elevation of the right diaphragm (2-3 intercostal), and it returned to normal 3 months after surgery.The CT review of 1 child with pulmonary wedge resection 1 month after surgery displayed a little residual lesion.Thoracoscopic lobectomy was significantly different from open surgery in terms of incision length[(2.0±0.5) cm vs. (10.0±0.5) cm], postoperative pain (the first time of getting out of bed)[(24.0±2.0) h vs. (48.0±1.5) h] and pulmonary complications (0 vs. 5 cases). The operative time of single lung ventilation was remarkably different from that of double lung ventilation in thoracoscopic lobectomy[(96.0±6.0) min vs. (118.0±7.0) min, t=106.501, P<0.001]. Compared with thoracoscopic lobectomy, thoracoscopic pulmonary segmentectomy has a long time and a large amount of blood loss. Conclusions:Complete thoracoscopic lobectomy (segmental) is the first choice for the treatment of congenital pulmonary airway malformation in children, with clear operative field, little trauma, less postoperative pain, quick recovery and fewer respiratory complications.Single-lung ventilation is beneficial for surgical completion.

Objective: To evaluate the relationship between bolus volume and hyoid displacement in dysphagia patients with nasopharyngeal carcinoma after radiation therapy. Methods: Twenty-three nasopharyngeal carcinoma patients with dysphagia were recruited and their swallowing of 3, 5, 10 and 20ml of liquid food was studied fluoroscopically. The vertical and horizontal displacement of the hyoid as well as its time in motion were measured, and the relationship between the bolus volume, hyoid displacement and time in motion time was evaluated. Results: The largest vertical displacement of the hyoid (1.01±0.65cm) was observed when swallowing a 10ml bolus. The hyoid showed the smallest average horizontal displacement (0.39±0.34cm), when swallowing a 3ml bolus. The average motion time of the hyoid was (2.11±0.65) seconds. It was shorter when swallowing a 10 or 20ml bolus than when dealing with a smaller one. Hyoid motion time was negatively correlated with the horizontal displacement of the hyoid bone, and the volume of a swallow was negatively correlated with the hyoid motion time but positively correlated with the penetration-aspiration scale score. Conclusion Bolus volume affects hyoid displacement and hyoid motion time in nasopharyngeal carcinoma patients with dysphagia after radiation therapy. For patients with a penetration-aspiration scale score of 5 or less, the optimum bolus volume is 5 to 10ml.

Objective To evaluate Aptima HPV E6 and E7 mRNA assay (Aptima HPV) combined with Aptima HPV 16 and 18 or 45 (18/45) genotype assay (Aptima HPV-GT) as a means of cervical cancer opportunistic screening. Methods From October 2016 to October 2017, a total of 23 258 women aged 25-65 years were enrolled in the physical examination center and gynecological clinic of Huzhou Maternity and Child Health Care Hospital. All the women had Aptima HPV tested, further Aptima HPV-GT testing for positive women and liquid-based thin layer cytology Thinprep cytologic test (TCT). Women with Aptima HPV (+) or ≥low-grade squamous intraepithelial lesion (LSIL) or obvious clinical symptoms (including vaginal bleeding after intercourse and watery, bloody vaginal discharge) were referred for colposcopy and further biopsy with or without endocervical curettage (ECC) if indicated. Expression of Aptima HPV, HPV 16 and HPV 18/45 with different cytological diagnostic groups and histological diagnosis groups were compared respectively. Sensitivity, specificity, positive predictive value and negative predictive value of Aptima HPV detection and TCT in identifying histological diagnosis of high-grade squamous intraepithelial lesion (HSIL) or worse (HSIL+) were compared. Results (1) The positive rates of Aptima HPV, HPV 16 and HPV 18/45 were 14.00% (3 257/23 258), 1.85% (430/23 258) and 0.86% (199/23 258) respectively.The positive rates of Aptima HPV, HPV 16 and HPV 18/45 increased with cytology grading in squamous epithelium [negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASCUS), LSIL, atypical squamous cell cannot exclude HSIL (ASC-H), HSIL and squamous cell carcinoma (SCC), all P=0.000)]. According to histology results, the positive rates of Aptima HPV, HPV 16 and HPV 18/45 increased with histology grading in squamous epithelium (normal cervical tissue, LSIL, HSIL and SCC, all P=0.000). The positive rate of Aptima HPV was significantly higher in HSIL+group than that in the LSIL or better (LSIL-) group [98.11% (311/317) vs 12.84% (2 946/22 941), P=0.000]. The positive rate of Aptima HPV-GT was significantly higher in HSIL+group than that in LSIL-group [58.36% (185/317) vs 1.91% (439/22 941), P=0.000]. (2) Compared with cytology, Aptima HPV resulted in significant higher sensitivity (98.11% vs 59.62%, P=0.000) and negative predictive value (99.97% vs 99.42%, P=0.000), significant lower specificity (87.16% vs 95.37%, P=0.000) and positive predictive value (9.55% vs 15.10%, P=0.000) when identified HSIL+. Conclusions Women with Aptima HPV positive, especially those with Aptima HPV-GT positive, are more likely to have histological diagnosis of HSIL+. Aptima HPV combined with Aptima HPV-GT is feasible as a means of cervical cancer opportunistic screening in tertiary hospitals.

Objective To investigate the distribution differences of human papillomavirus(HPV) genotypes in the patients with cervical intraepithelial neoplasia grade Ⅱ/Ⅲ and the patients with cervical cancer. Methods 260 cases of cervical cancer and 280 cases of cervical intraepithelial neoplasia grade Ⅱ /Ⅲ patients who were admitted to our hospital from January 2010 to October 2017 were selected and divided into cancer group and neoplasia group respectively. The HPV infection of patients was detected and typing identification was performed. The distribution differences of HPV genotypes in cancer group and neoplasia group were compared. The differences of genotypes of HPV infection in different types of tissues in cancer group were also compared. Results The negative constituent ratio of HPV infection in cancer group was lower than that in neoplasia group(P＜0. 05), and the constituent ratio of all the high-risk types in single infection was significantly higher than that in the neoplasia group(P＜0. 05). The constituent ratio of all the low-risk types in single infection was significantly similar to that in the neoplasia group(P＞0. 05). The constituent ratio of multiple infections in cancer group was significantly higher than that in neoplasia group(P＜0. 05); the constituent ratios of negative HPV infections and 18 types of single infections in cervical squamous cell carcinoma was much lower than that of cervical adenocarcinoma(P＜0. 05). The constituent ratios of 16 types of HPV and multiple infections in cervical squamous cell carcinoma were much higher than that of cervical adenocarcinoma(P＜0. 05). Conclusion The risk of HPV infection in cervical cancer patients is significantly higher than that of cervical intraepithelial neoplasia grade Ⅱ /Ⅲ, the infection rate of cervical cancer high-risk type HPV is higher, and the risk of multiple infections is also higher. The tissue types of cervical cancer can be determined according to HPV typing.

Objective The aim of the current study is to determine the clinical value of incidental internal mammary lymph node biopsy in free abdominal flap breast reconstruction using internal mammary vessels as recipient vessels and to investigate the risk factors of internal mammary lymph nodes metastasis. Methods The clinical data of all patients who underwent free abdominal flap breast reconstruction using internal mammary vessels as recipient vessels from November 2006 to December 2015 in the Department of Breast Surgery, Fudan University Shanghai Cancer Center were reviewed in the study. The incidence of internal mammary lymph node biopsy and the rate of metastasis were analyzed. Statistical analysis was conducted to evaluate the risk factors of internal mammary lymph node metastasis. Results A total of 113 patients met the inclusion criteria, 53 (46.9%) of whom had internal mammary lymph nodes harvested. Four of these were positive for metastatic disease, all in immediate breast reconstructions. The incidence of metastasis was 7.5% in patients who had successful internal mammary lymph node biopsies.The multi?variate Logistic regression analysis showed that invasive tumor size, tumor location and axillary lymph node metastasis were not risk factors for internal mammary lymph node metastasis ( P>0. 05 ) . Conclusions Internal mammary lymph nodes found incidentally during recipient vessel exposure may provide important information about internal mammary lymph node metastasis in free flap breast reconstruction. This approach for internal mammary lymph node biopsy reveals an appreciable success rate and is convenient in clinical practice. The size of invasive tumor and the axillary lymph node metastasis are probably associated with internal mammary lymph node positivity.

Objective The aim of the current study is to determine the clinical value of incidental internal mammary lymph node biopsy in free abdominal flap breast reconstruction using internal mammary vessels as recipient vessels and to investigate the risk factors of internal mammary lymph nodes metastasis. Methods The clinical data of all patients who underwent free abdominal flap breast reconstruction using internal mammary vessels as recipient vessels from November 2006 to December 2015 in the Department of Breast Surgery, Fudan University Shanghai Cancer Center were reviewed in the study. The incidence of internal mammary lymph node biopsy and the rate of metastasis were analyzed. Statistical analysis was conducted to evaluate the risk factors of internal mammary lymph node metastasis. Results A total of 113 patients met the inclusion criteria, 53 (46.9%) of whom had internal mammary lymph nodes harvested. Four of these were positive for metastatic disease, all in immediate breast reconstructions. The incidence of metastasis was 7.5% in patients who had successful internal mammary lymph node biopsies.The multi?variate Logistic regression analysis showed that invasive tumor size, tumor location and axillary lymph node metastasis were not risk factors for internal mammary lymph node metastasis ( P>0. 05 ) . Conclusions Internal mammary lymph nodes found incidentally during recipient vessel exposure may provide important information about internal mammary lymph node metastasis in free flap breast reconstruction. This approach for internal mammary lymph node biopsy reveals an appreciable success rate and is convenient in clinical practice. The size of invasive tumor and the axillary lymph node metastasis are probably associated with internal mammary lymph node positivity.

Objective To explore the clinical significance of human papillomavirus L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women. Methods From November 2012 to June 2013,386 high-risk HPV positive (detected by hybrid capture Ⅱ) cases were enrolled as eligible women from Huzhou Maternity&Child Care Hospital and Women′s Hospital,School of Medicine, Zhejiang University. All eligible women underwent liquid-based cytology (ThinPrep) followed by colposcopy. Biopsies were taken if indicated. Cervical exfoliated cells were collected for HPV L1 capsid protein detection by immunocytochemistry. Expression of HPV L1 capsid protein in groups with different histological diagnosis were compared, and the role of HPV L1 capsid protein detection in cervical exfoliated cells in cervical lesions screening was accessed. Results Total 386 enrolled eligible women were finally diagnosed histologically as follwed:162 normal cervix, 94 low-grade squamous intraepithelial lesion (LSIL), 128 high-grade squamous intraepithelial lesion (HSIL) and 2 squamous cervical cancer (SCC). The positive expression rate of HPV L1 in HSIL+(HSIL or worse) group was significantly lower than that in LSIL-(LSIL or better) group (19.2% vs 66.4%,P=0.000). While identifying HSIL+ in HPV positive cases and compared with cytology, HPV L1 detection resulted in significant higher sensitivity (80.77%vs 50.77%,P=0.000) and negative predictive value (NPV;87.18% vs 76.47%,P=0.004), significant lower specificity (66.41% vs 81.25%,P=0.000),and comparable positive predictive value (PPV;54.97% vs 57.89%, P=0.619). To identify HSIL+in HPV-positive/cytology-negative women, the sensitivity, specificity, PPV, and NPV of HPV L1 detection were 87.50%, 61.54%, 41.18%, and 94.12%respectively, while 80.00%, 86.36%, 80.00%and 86.36%respectively in HPV-positive/atypical squamous cell of undetermined significance(ASCUS)women. Conclusions HPV L1 capsid detection in cervical exfoliated cells have a role in cervical lesions screening in high-risk HPV positive women, and may be a promising triage for high-risk HPV-positive/cytology-negative or ASCUS women.