Objective To select and optimize the process parameters of SARS-CoV-2 F61 affinity chromatography by the screening and optimization experiment of Design of Experiment(DoE),in order to obtain the optimal process conditions.Methods Eight process parameters that may affect the experimental response results in affinity chromatography were selected and their level ranges were determined.DoE screening test was used to perform 8 factor 2 level screening tests on the selected process parameters.The response values were detected and the mathematical model was fitted by statistical software.Three key process parameters significantly affecting the key quality attributes were obtained by Pareto diagram analysis(P < 0.05).Then DoE response surface method(RSM)was selected to optimize the key process parameters.First,the full factorial experiment design was completed,the response results were detected to fit the mathematical model,and the bending term P value was analyzed to judge the range of significant factors in the optimal range(bending P < 0.05),then according to the sequential complement of the central composite face-centered design(CCF)experiment,through the detection of the response results to fit the response surface model,the range of optimal conditions was obtained,and the stability of the optimal parameters was finally verified by repeated experiments.Results In the screening experiment,it was found that the significant factors affecting F61 in the affinity chromatography stage were elution buffer pH,elution buffer salt concentration,leaching buffer salt concentration and equilibrium buffer salt concentration.The optimal conditions of key process parameters in affinity chromatography were obtained by CCF.When the pH of elution buffer was 3.2,the elution buffer salt concentration was 0.07 mol/L NaCl,and the leaching buffer salt concentration was 0.31 mol/L NaCl,the yield of F61 reached 95.25%,the residual amount of host cell protein(HCP)was 97.33 ppm,and the monomer purity of the sample was98.51% at the affinity chromatography stage.Conclusion Different types of DoE methods were used to screen and optimize the process parameters of F61 affinity chromatography stage,and the optimal process conditions were obtained,which lays a foundation for the esta-blishment of F61 purification process.

Objective To evaluate the effects of the modified opration of benign tumor in parotid gland on the prevention of Frey syndrome.Methods the Parotid benign tumor resection in 28 cases were selected,according to the different treatment methods were divided into observation group of 12 cases,16 cases of the control group.The modified surgical patients in the observation group,two flap method,preservation,preservation of parotid masseter fascia in the branches of the great auricular nerve and parotid region resection,the control group was treated by routine tumors of the parotid gland and parotid superficial lobectomy or resection.Patients were followed up for 6 months to 1 years,underwent minor test,the incidence of Frey syndrome in 2 cases were recorded,postoperative incidence of Frey syndrome were compared.Results The incidence of Frey syndrome in the observation group was significantly reduced compared with the control group.The difference was statistically significant (χ2 =7.479,P <0.05 ). Conclusion The effects of modified opetation combined with secondry disc method in benign tumor of parotid gland on the prevention of Frey syndrome has obvious advantages comparing with the control.

Objective Implanting surgical meshes to support breast in periareoler mammaplasty shows an excellent prospect.But shrinking of the mesh and the adhesion formation affected the postoperative shape and palpation of the breast.The aim of this study was to compare biocompatible properties of four different polypropylene meshes (Prolene(R) ; Vypro Ⅱ(R) ; Premilene Mesh (R) and Premilene Mesh LP(R)) and to discuss the inclusion criteria of meshes in this technique.Methods The study was carried out in pigs.Four different polypropylene meshes were implanted subcutaneously in belly.90 days after implantation the mesh samples were explanted,and complications,foreign-body reaction,collagen fiber and elastic fiber formation were observed,as well as ultrastructures for the biocompatible properties.Results The biocompatible properties of each mesh were different.The foreign-body reactions determined the biocompatible properties,and affected the postoperative shape greatly.Conclusions Premilene Mesh LP o as a newer low-weight polypropylene mesh,has superior properties to the others.

Objective To observe the distribution of sentinel nodes(SN) in gastric cancer,and investigate their clinical significance.Methods The extent and features of intraoperatively stained sentinel nodes in 288 cases of gastric cancer were analyzed retrospectively.Methylene blue was injected intraoperatively into the(subseral) muscle and submucosa of the normal gastric wall adjacent to the neoplasms,then lymphadenography was performed,and the nodes were resected for histopathological examination.Results The pathological(diagnosis) of the 288 cases with gastric cancer was T_1 in 102 cases,T_2 106 cases and T_3 60 cases.Stained nodes were detectable in 270 cases(success rate: 93.8%).One hundred and two cases were found to have lymph node metastasis;66 cases had metastasis in both SNs and non-SNs;18 cases in SNs alone;and 18 cases in non-SNs alone.Conclusions Sentinel lymph nodes can accurately predict the lymph node metastatic status of gastric cancer patients during operation.Intraoperative sentinal lymph node mapping can avoid routine lymph node dissection in patients with gastric cancer with negative lymph node metastasis.

Objective To investigate the value of intervention bundles in enteral nutrition for patients with esophageal cancer.Methods From October 2014 to September 2015,226 patients with esophageal squamous cell carcinoma were collected.From April 2015 to September 2015,109 patients(intervention group)were treated with intervention bundles during perioperative period,and from October 2014 to March 2015,117 patients(control group)were treated by routine intervention.Results The albumin,prealbumin,and transferrin showed no significant difference(all P>0.05)before treatment but were significantly different on the eighth day[albumin:(38.2±3.5)g/L vs.(36.3±4.8)g/L,P=0.001;prealbumin:(126.7±52.8)g/L vs.(72.9±42.3)g/L,P=0.001;transferrin:(2.9±1.2)g/L vs.(2.1±1.6)g/L,P=0.001].The incidence of complications was 11.01％(12/109)in intervention group and 21.37％(25/117)in control group(X2=4.422,P=0.035).In addition,the postoperative exhaust time[(52.8±10.9)h vs.(58.7±14.3)h,P=0.001],time to the removal of chest drainage tube[(3.5±0.9)d vs.(4.8±1.3)d,P=0.001],postoperative hospital stay[(11.2±1.3)d vs.(12.1±1.5)d,P=0.001],and hospital costs[(37±4)thousand yuan vs.(39±5)thousand yuan,P=0.004] were also significantly shorter or smaller in the intervention group.Conclusions Intervention bundles is clinically valuable in the early enteral nutrition for patients with esophageal cancer.It can improve the nutritional status of patients,reduce complications,and improve the clinical outcomes.

Objective To investigate the effect of all-in-one nursing intervention on esophageal carcinoma patients with bad emotion and immune function. Methods From the October, 2012 to the October, 2013, 102 patients with radical operation of esophageal carcinoma were divided into all-in-one nursing intervention group (research group, 52cases) and routine nursing group (control group, 50 cases) according to the random number table method.Two groups of patients were evaluated before and after operation of anxiety and depression emotion scores, with the Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) .At the same time, two groups of patients were detected the stress and immune index before and after the operation. Results Before operation, two groups of patients with anxiety, depression, stress reaction and immune index were not significantly different (P>0.05).After operation, the study group patients with anxiety and depression scores were significantly lower than those in the control group [(34.6+6.7) points vs. (44.8+7.5) points, (33.7+6.9) points vs. (40.6+7.3) points], the difference was statistically significant (t values were 7.234, 4.902, P<0.05). Eighth days after operation, patients in the control group FBG, CRP and Cor levels were significantly higher in the study group [(6.98±1.12)mmol/L vs.(6.09±0.85)mmol/L,(18.11±5.93) mg/L vs.(9.24±3.22)mg/L,(293.98±69.09)nmol/L vs.(221.97±67.94)nmol/L],the difference was statistically significant (t values were 4.532, 9.336, 5.307, P<0.05). Eighth days after operation, two groups of patients with IgA, IgM were higher than first days after operation [(2.41±0.46)g/L vs.(1.89±0.56)g/L,(2.51±0.58) g/L vs.(1.35±0.67)g/L and (2.36±0.53)g/L vs.(1.82±0.46)g/L, (2.34±0.62)g/L vs. (1.37±0.64) g/L], the difference was statistically significant (t values were 5.174, 9.439, 5.549, 7.850, P<0.05).The total number of patients blood lymphocytes (LYM) and cytokines (CD3, CD4, CD8, CD4/CD8) were significantly higher than the control group [(2.87±0.65)×109/L vs.(1.98±0.58)×109/L, 0.62±0.10 vs. 0.57±0.07, 0.41±0.08 vs. 0.35 ±0.08, 0.24 ±0.04 vs. 0.26 ±0.05, 2.06 ±0.44 vs. 1.65 ±0.46], the difference was statistically significant (t values were 7.286, 2.915, 6.942, 2.225, 4.601, P<0.05). Conclusion All-in-one nursing intervention can significantly improve the anxiety and depression in patients with esophageal cancer in bad mood, enhance the immune function of patients, conducive to the rehabilitation of patients.

Objective: To investigate the clinical and pathological features, diagnosis and treatment of primary vulvar Paget disease (VPD) , and analyze the related factors that may affect the recurrence. Methods: A retrospective study was carried out on 36 patients diagnosed as VPD pathologically from January 1983 to December 2017 at Peking Union Medical College Hospital, Chinese Academy of Medical Sciences. The clinical and pathological features, diagnosis, treatment and prognosis and the factors influencing recurrence rate of VPD were analyzed. Results: (1) Totally 94% (34/36) of VPD occurred in postmenopausal women. Pruritus was counted 86% (31/36) of the main complaint. Lesions of vulvar were main symptom which had no specificity, acting as ulcer (67%, 24/36) , erythema (50%, 18/36) , depigmentation (42%, 15/36) , sclerosis (31%, 11/36) , and pigmentation (17%, 6/36) . The lesions invaded labium majus (97%, 35/36) , sometimes labium minus (53%, 19/36) , clitoris (28%, 10/36) , perianal (25%, 9/36) , orificium vaginae (3%, 1/36) , and meatus urinarius (3%, 1/36) . Approximately 19% (7/36) of VPD coexisted with intraepithelial neoplasia or adenocarcinoma of vulvar or other part of body. (2) Diagnosis and treatment: diagnosis was confirmed histologically by biopsy or pathologies after surgery, and immunohistochemical results were helpful for differential diagnosis. Surgery was the mean treatment method, 34 of all the 36 patients (94%, 34/36) underwent surgery for at least once, while 2 patients (6%, 2/36) were performed non-operative treatment. The surgical treatment included excision of focus, wide local excision, simple vulvectomy, and extensive vulvectomy. The non-operative treatment included radiotherapy, chemotherapy, laser, photodynamic therapy, and so on. (3) Prognosis: among 36 VPD patients, 4 were lost to follow-up with a 89% (32/36) follow-up rate. Median follow-up was 35.3 months (range,1 month to 31 years) . During the follow-up period, 2 patients were unable to judge whether they will relapse for the follow-up time did not reach half a year, 8 cases were unsuccessful operation, 20 cases succeeded, the achievement ratio was 71% (20/28) . Nine of twenty cases relapsed, the recurrence rate was 45% (9/20) . The median recurrence time was 14 months after operation. One patient of the 32 followed-up patients died, the mortality rate was 3% (1/32) . (4) The related factors affected the recurrence of VPD: t test was applied to the analysis of patients′ age, rank test was used in the statistics of the time of confirmed diagnosis, the length and thickness of the resection focus. Fisher test was used to calculate whether the focus were limited to the epidermis, type of surgical procedures, distance between the margin and the focus, whether tumor cells infiltrated the margin. The results showed that none of the above terms in the first operation had significant contribution to recurrence (all P>0.05) . Conclusions VPD may be a low potential malignancy, which could slowly progress into deep invasive disease. VPD is often associated with intraepithelial neoplasia or primary tumors of the vulva or somewhere else. Operations is the first choice for VPD, but consider for its high recurrence rate after operation, close follow-up should be strongly suggested.

OBJECTIVE: To determine the influence of different registration scopes and methods on kilo-voltage cone beam CT (kVCBCT) image and CT image and to estimate the appropriate clinical target volume (CTV)-to-planned target volume (PTV) margin for central-type non-small cell lung cancer in image-guided radiation therapy (IGRT). METHODS: Twenty-six patients with central-type non-small cell lung cancer (NSCLC) who received IGRT were enrolled in this study and totally 104 flames of kVCBCT scanning acquired before radiotherapy were analyzed. First, registrations were performed by gray plus manual match and were compared among different registration scopes of tumor and tumor plus vertebra. Then, the results of registration as well as time cost using tumor plus vertebra as registration scope were compared among different registration methods of gray plus manual match and bone plus manual match. At last, 200 registrations using tumor plus vertebra as the registration scope performed by bone plus manual match were recorded and the CTV-to-PTV margin was calculated. RESULTS: There was no significant difference in X, Y and Z translational and rotational movements between the registration scopes of tumor and tumor plus vertebra when gray plus manual match was used as the registration method (P>0.05). The registration results of gray plus manual match and bone plus manual match were equal when tumor plus vertebra was used as the registration scope (P>0.05), but the time cost of bone plus manual match [(1.9±0.3) min] was shorter than that of gray plus manual match [(3.1±0.2) min]. The CTV-to-PTV margins were 5.3, 4.9 and 5.7 mm in X, Y, and Z directions respectively. CONCLUSION For central-type NSCLC, tumor and vertebra can be used as registration scope and the bone plus manual match is suggested in IGRT. To avoid errors, we suggest a CTV-to-PTV margin of 6 mm.
Aged ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Cone-Beam Computed Tomography ; Female ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Patient Positioning ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Image-Guided ; methods

One hundred and eighty three patients with esophageal cancer admitted from September 2015 to September 2016 were randomly divided into two groups:91 patients received clinician-involving pre-discharge and postoperative follow-up health education (study group) and 92 patients received traditional health education (control group).The quality of life and the compliance rate of comprehensive treatment within 3 months after discharge were evaluated and compared between two groups.The overall scores of quality of life in study group was significantly better than those of the control group (P＜ 0.05),and the compliance rate of comprehensive treatment after discharge in study group was significantly higher than that of control group [82.4％(75/91) vs.67.4％(62/92),x2=5.49,P=0.02].It is suggested that clinician participating in the pre-discharge and follow-up health education can improve the quality of life of patients,and improve the compliance of comprehensive treatment after discharge.

Objective To investigate the biological function of long non-coding RNA(LncRNA)LINC01137 in immune escape of non-small cell lung cancer(NSCLC)cells and its potential regulatory mechanisms.Methods The blood samples of 24 healthy volunteers and 24 NSCLC patients were collected.The tumor tissues and paracancerous tissues of 24 NSCLC patients were collected,and the levels of LINC01137 were detected.The binding sites of LINC01137 and miR-22-3p were predicted by Starbase database and verified by the luciferase reporter gene analysis.A549 cells were transfected with exosomes derived from A549 cells and/or sh-LINC01137 interference sequence to detect cell proliferation and invasion.The supernatant of A549 cells were collected to culture CD8+T cells,and the levels of CD8+T cell exhaustion markers,including interfereron-γ(IFN-γ),tumor necrosis factor-α(TNF-α),granzyme B and interleukin-2(IL-2),and the percentage of PD-1+Tim3+CD8+T cells were detected.CD8+T cells were transfected with exosomes and/or miR-22-3p mimics to detect the protein level of PD-1.Results The expression of LINC01137 in tumor tissues of patients with NSCLC was increased compared with paracancerous tissues(3.357±0.548 vs 1.011±0.371),while the expression of LINC01137 in peripheral blood of patients with NSCLC was increased compared with healthy volunteers(3.216±0.342 vs 1.007±0.313),with statistically significant differences(t=-17.367,-17.147,all P<0.001).There was a positive correlation between the expression of LINC01137 in tumor tissue and peripheral blood(r=0.755,P<0.05).LINC01137 was significantly enriched in exosomes derived from A549 cells.Compared with Exo+sh-NC group,the cell viability(65.85%±4.71%vs 100.15%±11.93%)and cell invasion(21.46%±3.48%vs 43.12%±1.44%)in Exo+sh-LINC01137 group were decreased,and the differences were statistically significant(t=4.630,9.953,all P<0.01).The expression of LINC01137 in peripheral blood of NSCLC patients was negatively correlated with the percentage of CD8+T cells(r=-0.520,P<0.05).Compared with Exo+sh-NC group,the IFN-γ(3 865.31±543.85 pg/ml vs 1 786±105.98 pg/ml),TNF-α(4 631.93±510.71 pg/ml vs 1 973.24±379.62 pg/ml),Granzyme B(3 876.49±312.43 pg/ml vs 1 879.43±287.58 pg/ml),and IL-2 mRNA levels(3.286±0.437 vs 1.015±0.314)were increased,and the percentage of PD-1+Tim3+CD8+T cells(7.68%±2.18%vs 18.95%±3.21%)was decreased in Exo+sh-LINC01137 group,with statistical significances(t=-6.497,-7.237,-8.146,-7.310,5.021,all P<0.01).Our results showed that miR-22-3p was the target gene of LINC01137.Compared with Exo+NC mimic group,the level of PD-1 protein in Exo+miR-22-3p group(0.384±0.087 vs 1.003±0.147)was significantly decreased,and the difference was statistically significant(t=6.277,P<0.01).Conclusion The expression of LINC01137 was significantly up-regulated in tumor tissues and plasma of NSCLC patients.Exosomes LINC01137 derived NSCLC cell induces CD8+T cell exhaustion by targeting miR-22-3p and inhibiting its expression,and thus promoting NSCLC cell immune escape.