Objective To enhance the standardization of surgical operations,shorten the learning curve,and reduce surgical complications by summarizing common errors and proposing solutions for beginners during pre-study of cavity construction in non-inflatable axillary approach laparoscopic thyroid surgery.Methods A retrospective analysis was made on 210 patients who underwent noninflatable axillary approach luminal thyroid surgery at the Department of Head and Neck Surgery,Zhejiang Provincial People's Hospital,between January 2022 and June 2023.Among them,150 patients were luminalized by the practitioners in the practice group,while 60 patients were luminalized by the practitioners in the skilled group.The occurrence of nine common errors during cavity construction,such as supraclavicular nerve injury and scapulohumeral muscle injury,was assessed and compared between the two groups.Results There was no statistically significant difference in age,gender,tumor diameter,tumor location or pathology type between the two groups(P＞0.05).The staff in the practice group had a significant difference in supraclavicular cutaneous nerve injury(19.33％vs.3.33％),scapulohumeral muscle injury or disarticulation(16.00％vs.0),anterior cervical band of free excess(24.00％vs.3.33％),pectoralis major muscle and supraclavicular fascia injury(16.00％vs.5.00％),external jugular vein and genuine branch injury(8.00％vs.0),excessive freeing of the sternoclavicular joint(7.33％vs.0),and incorrect entry of the sternocleidomastoid muscle gap(8.67％vs.0)were significantly higher in incidence than the employees in the skilled group(P＜0.05).There was no statistically significant difference between employees in the practice group and those in the skilled group in terms of injury to the internal jugular vein and its geniculate branches(4.00％vs.0),and incorrect entry of the anterior cervical strap muscles into the hierarchy(2.00％vs.0)(P＞0.05).Conclusion The incidence of damage to the surrounding tissue was significantly higher in the novice group than in the expert group during endoscopic thyroid surgery for space construction.A solid anatomical foundation and proficient endoscopic surgical technique serve as fundamental prerequisites for achieving a successful space system construction.By summarizing common challenges and providing corresponding solutions encountered during endoscopic thyroid surgery,this study further enhances and refines the learning system for total endoscopic thyroid procedures.

Analyzing the distribution characteristics of allergen sIgE in the serum of patients with respiratory and skin mucosal diseases in Shanghai City, and to provide epidemiological characteristics and diagnostic basis for the prevention and treatment of allergic respiratory and dermo-mucous diseases in Shanghai City. Adopting cross-sectional research, a total of 3 822 patients who received treatment in Tongren Hospital, Shanghai Jiao Tong University School of Medicine from July 2022 to July 2023 due to respiratory diseases or skin and dermo-mucous symptoms were included. Among them, there were 1 456 males and 2 366 females, with an age range of 1-97 years old. The median age (interquartile range) was 33 (27, 44) years old. The sIgE was detected by using immunoblotting. Statistical analysis was conducted using SPSS 22.0 software, and the comparison of count data (rates) between groups was conducted using χ 2 test. The results showed that a total of 3 377 (88.4%) cases among 3 822 patients were at least one allergen sIgE positive, and 72.9% (2 788/3 822) of them were multiple allergies sIgE positive. The top five allergen sIgE positive rates were Dermatophagoides pteronyssinus (37.9%, 1 447/3 822), Dermatophagoides farinae (32.1%, 1 225/3 822), milk (31.7%, 1 211/3 822), fungi (28.3%, 1 080/3 822), and Blomia tropicdis (23.8%, 909/3 822), with only milk was a kind of food allergen. The highest positive rates within the respiratory system disease group or dermo-mucous disease group were also these five allergens, without any difference in disease categories. The positive rates of cat dandruff, Humulusscandens, and juniper/birch in the respiratory system disease group were significantly higher than those in the skin and mucous membrane disease group, while the positive rates of shrimp/crab were relatively low (11.3% vs 14.9%, χ 2=9.616, P=0.002). Whether in the respiratory system disease group or the dermo-mucous disease group, the positive rates of Dermatophagoides pteronyssinus in male patients were significantly higher than those of females(42.6% vs 35.7%,41.0% vs 34.4%), with statistical significance ( χ 2=12.515, P<0.001; χ 2=5.143, P=0.023), And the three allergens, Dermatophagoides farinae, cat dander, and egg white allergens are also characterized by this feature.In addition, the positive rates of milk(33.8% vs 30.1%, χ 2=3.911, P=0.048), shrimp/crab(13.2% vs 10.0%, χ 2=6.423, P=0.014) in the respiratory system disease group were higher in males than in females, while in the dermo-mucous disease group, dog dander(20.5% vs 14.6%, χ 2=6.726, P=0.010) and peanuts/soybeans(10.5% vs 6.9%, χ 2=4.698, P=0.030) showed this phenomenon. In both the respiratory system disease group and the dermo-mucous disease group, there were 6 types of inhaled allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicdis, cat dandruff, dog dander, fungi) and 4 types of food allergens (egg yolks, egg white allergens, milk, shrimp/crab). However, the positivity rate of Aspergillus fumigatus (7.2% vs 9.3% vs 10.5% vs 15.7%, χ 2=10.996, P=0.012)in the respiratory disease group and cockroaches(4.2% vs 11.3% vs 9.6% vs 16.4%, χ 2=10.237, P=0.017) in the skin and mucosal disease group was the lowest in the underage group. There are seasonal differences in the positivity rates of allergens, with most allergens having significantly higher positivity rates in summer and autumn. In conclusion, the most common allergens sIgE positive in patients with respiratory and dermo-mucous diseases in Shanghai City are Dermatophagoides pteronyssinus, Dermatophagoides farinae, milk, fungi, and Blomia tropicdis. The trend of allergen sIgE prevalence in the two major categories of diseases is basically consistent. Allergen sIgE distribution varies among patient populations of different gender, age or season, and clinical prevention and treatment can be based on the results of serum allergen testing.

Analyzing the distribution characteristics of allergen sIgE in the serum of patients with respiratory and skin mucosal diseases in Shanghai City, and to provide epidemiological characteristics and diagnostic basis for the prevention and treatment of allergic respiratory and dermo-mucous diseases in Shanghai City. Adopting cross-sectional research, a total of 3 822 patients who received treatment in Tongren Hospital, Shanghai Jiao Tong University School of Medicine from July 2022 to July 2023 due to respiratory diseases or skin and dermo-mucous symptoms were included. Among them, there were 1 456 males and 2 366 females, with an age range of 1-97 years old. The median age (interquartile range) was 33 (27, 44) years old. The sIgE was detected by using immunoblotting. Statistical analysis was conducted using SPSS 22.0 software, and the comparison of count data (rates) between groups was conducted using χ 2 test. The results showed that a total of 3 377 (88.4%) cases among 3 822 patients were at least one allergen sIgE positive, and 72.9% (2 788/3 822) of them were multiple allergies sIgE positive. The top five allergen sIgE positive rates were Dermatophagoides pteronyssinus (37.9%, 1 447/3 822), Dermatophagoides farinae (32.1%, 1 225/3 822), milk (31.7%, 1 211/3 822), fungi (28.3%, 1 080/3 822), and Blomia tropicdis (23.8%, 909/3 822), with only milk was a kind of food allergen. The highest positive rates within the respiratory system disease group or dermo-mucous disease group were also these five allergens, without any difference in disease categories. The positive rates of cat dandruff, Humulusscandens, and juniper/birch in the respiratory system disease group were significantly higher than those in the skin and mucous membrane disease group, while the positive rates of shrimp/crab were relatively low (11.3% vs 14.9%, χ 2=9.616, P=0.002). Whether in the respiratory system disease group or the dermo-mucous disease group, the positive rates of Dermatophagoides pteronyssinus in male patients were significantly higher than those of females(42.6% vs 35.7%,41.0% vs 34.4%), with statistical significance ( χ 2=12.515, P<0.001; χ 2=5.143, P=0.023), And the three allergens, Dermatophagoides farinae, cat dander, and egg white allergens are also characterized by this feature.In addition, the positive rates of milk(33.8% vs 30.1%, χ 2=3.911, P=0.048), shrimp/crab(13.2% vs 10.0%, χ 2=6.423, P=0.014) in the respiratory system disease group were higher in males than in females, while in the dermo-mucous disease group, dog dander(20.5% vs 14.6%, χ 2=6.726, P=0.010) and peanuts/soybeans(10.5% vs 6.9%, χ 2=4.698, P=0.030) showed this phenomenon. In both the respiratory system disease group and the dermo-mucous disease group, there were 6 types of inhaled allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicdis, cat dandruff, dog dander, fungi) and 4 types of food allergens (egg yolks, egg white allergens, milk, shrimp/crab). However, the positivity rate of Aspergillus fumigatus (7.2% vs 9.3% vs 10.5% vs 15.7%, χ 2=10.996, P=0.012)in the respiratory disease group and cockroaches(4.2% vs 11.3% vs 9.6% vs 16.4%, χ 2=10.237, P=0.017) in the skin and mucosal disease group was the lowest in the underage group. There are seasonal differences in the positivity rates of allergens, with most allergens having significantly higher positivity rates in summer and autumn. In conclusion, the most common allergens sIgE positive in patients with respiratory and dermo-mucous diseases in Shanghai City are Dermatophagoides pteronyssinus, Dermatophagoides farinae, milk, fungi, and Blomia tropicdis. The trend of allergen sIgE prevalence in the two major categories of diseases is basically consistent. Allergen sIgE distribution varies among patient populations of different gender, age or season, and clinical prevention and treatment can be based on the results of serum allergen testing.

Objective:To investigate the effectiveness, safety, and advantages of modified radical neck dissection by gasless unilateral axillary approach (GUA-MRND) in the surgical management of selected patients with papillary thyroid cancer.Methods:We retrospectively analyzed patients with papillary thyroid cancer who underwent GUA-MRND (endoscopic group, n=16) versus unilateral open modified radical neck dissection (MRND) (open group, n=32) during the period from Jan. 2019 to Jun. 2021, including the differences in surgical efficiency, complication rate, and incisional satisfaction.Results:Compared MRND with GUA-MRND, the patients were younger ( P<0.05) , operative time and postoperative drainage anterior ( P<0.01) were slightly inferior in the latter, but it had obvious advantages in cervical swallowing discomfort and incision satisfaction evaluation ( P<0.05) . There was no significant difference in the incidence of temporary recurrent laryngeal nerve injury, intraoperative and postoperative bleeding, hematoma, infection, lymphatic or chylous leakage and supraclavicular numbness after surgery ( P>0.05) . The number of dissected lymph nodes in area II in the GUA-MRND was lower ( P<0.05) , but it was significantly higher ( P<0.01) in area III. And the average regional cleaning efficiency in the GUA-MRND was level Ⅲ (35.5%) , level Ⅵ (28.59%) , level Ⅳ (23.21%) , level Ⅱ (7.18%) and level Ⅴ (7.12%) , suggested that GUA-MRND had higher efficacy for level III, level Ⅵ and Level IV. Conclusion:GUA-MRND is safe, effective, and has high cosmetic satisfaction in the treatment of selected patients with lateral cervical lymph node metastases from papillary thyroid cancer.

Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.

Objective:To investigate the effects of Anhydroicaritin (AHI) , an isopentenylated flavo-noid compound, on proliferation, migration and apoptosis of human hepatocarcinoma cell line MHCC-97H.Methods:Human hepatocarcinoma cell line MHCC-97H and human normal liver cell line L02 were cultured in vitro. MHCC-97H cells were treated with 0, 20, 40, 80, 120, 160, 200 μg/ml of AHI respectively and L02 cells were treated with 0, 25, 50, 100, 150, 200, 400, 500 μg/ml of AHI respectively. CCK-8 and clone formation assay were used to detect cell proliferation. Scratch test was used to explore cell migration ability. Hoechst33342 assay and flow cytometer were used to detect cell apoptosis. The expressions of apoptosis-related proteins were detected by Western blotting. Results:The cell viabilities of MHCC-97H cells treated with 0, 20, 40, 80, 120, 160, 200 μg/ml of AHI for 24 h were (100.00±0.00) %, (97.41±2.10) %, (96.58±3.23) %, (87.72±4.85) %, (78.33±3.76) %, (56.97±2.61) % and (15.25±2.51) % respectively, and there was a statistically significant difference ( F=429.20, P<0.001) . There were statistically significant differences between 0 μg/ml and 80, 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . The cell viabilities of L02 cells treated with 0, 25, 50, 100, 150, 200, 400, 500 μg/ml of AHI for 24 h were (100.00±0.00) %, (96.82±3.79) %, (95.36±3.43) %, (90.79±5.75) %, (77.67±5.66) %, (63.98±5.22) %, (34.22±4.01) % and (33.84±4.41) % respectively, and there was a statistically significant difference ( F=233.20, P<0.001) . There were statistically significant differences between 0 μg/ml and 100, 150, 200, 400, 500 μg/ml of AHI treatment (all P<0.05) . The 24 h half maximal inhibitory concentration (IC 50) value of AHI treated L02 cells was (300.20±17.10) μg/ml, which was significantly higher than that of MHCC-97H cells [ (158.60±5.50) μg/ml], and there was a statistically significant difference ( t=13.65, P<0.001) . The cell clone numbers of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml of AHI for 24 h were 1 993.00±46.29, 1 355.00±54.84, 998.33±21.03 and 218.33±35.95 respectively, and there was a statistically significant difference ( F=954.80, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . The healing rates of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml of AHI for 24 h were (51.68±1.93) %, (16.04±0.73) %, (8.88±0.31) % and (-6.94±0.46) % respectively, and there was a statistically significant difference ( F=1 616.00, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . Hoechst33342 experiment showed that MHCC-97H cells treated with 0 μg/ml AHI showed uniform dark blue with a complete nuclear state under inverted microscope. Compared with 0 μg/ml AHI treated cells, cells in the 120, 160, 200 μg/ml AHI treatment groups wrinkled and broken, and nuclei were also morphologically abnormal, with some nuclei stained bright blue, and the situation became more obvious with increasing dose. The apoptosis rates of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml AHI for 24 h were (10.51±0.56) %, (42.23±0.87) %, (61.92±0.52) % and (72.05±0.74) % respectively, and there was a statistically significant difference ( F=4 677.00, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . There were statistically significant differences among the different expression levels of Bax, Cleaved Caspase-3/Caspase-3, Cleaved Caspase-9/Caspase-9, and Bcl-2 proteins in MHCC-97H cells of 0, 120, 160, and 200 μg/ml of AHI treatment ( F=30.43, P<0.001; F=212.80, P<0.001; F=475.30, P<0.001; F=10.75, P=0.004) . The Bax protein expression of 160 and 200 μg/ml was significantly increased than that of 0 μg/ml AHI (both P<0.001) . The Cleaved Caspase-3/Caspase-3, Cleaved Caspase-9/Caspase-9 protein expressions of 120, 160 and 200 μg/ml were significantly increased than those of 0 μg/ml AHI (all P<0.001) . The Bcl-2 protein expression of 120, 160, 200 μg/ml was significantly decreased compared with that of 0 μg/ml AHI (all P<0.05) . Conclusion:AHI can inhibit the proliferation and migration of hepatocellular carcinoma cell line MHCC-97H, and promote its apoptosis.

Objective : The live cell application was used to observe the process of phagocytosis of endometrial cancer cells by macrophages, and flow cytometry was used to detect the effects of macrophages engulfing tumor cells on activation of cytotoxic T cell. Methods : Ishikawa cells and THP1-induced macrophages were labeled with CFSE fluorescent probe and CD11 b respectively, and then mixed and seeded on a glass imaging dish.The live cell application was performed to record the phagocytosis of Ishikawa cells by macrophages within 120 minutes.Flow cytometry was used to detect the effect of macrophages engulfing tumor cells on activation of cytotoxic T cell. Results : The green fluorescence of Ishikawa cells was taken up by macrophages after the co-cultured two types of cells were in contact with each other, and macrophages were able to engulf multiple Ishikawa cells continuously.Macrophages that engulfed Ishikawa cells could induce activation of cytotoxic T cell. Conclusion The live cell application was successfully conducted to construct an in vitro model of phagocytosis of tumor cells by macrophages, which provided a feasible experimental method for detecting the dual killing process of macrophages and T cells on tumor cells.

Objective:To compare the postoperative neck pain, discomfort and swallowing difficulty in patients with papillary thyroid cancer (PTC) after conventional open thyroidectomy (COT) and endoscopic thyroidectomy using the gasless unilateral axillary approach (ET-UA) .Methods:The study included 117 female patients with PTC who underwent unilateral thyroid lobectomy plus central neck dissection using COT ( n=68) or ET-UA ( n=48) performed by the same experienced surgeon. Subjective neck pain, discomfort and swallowing alterations were assessed by questionnaire 3 day and 6 months postoperatively. Surgical scar and cosmetic satisfaction evaluation were implemented 6 months postoperatively. SPSS 20.0 was used for statistical analysis, the rate or composition ratio was compared by the χ2 test or the exact probability method, and the significance test of the difference in sample means was tested by analysis of variance. Results:Three days after surgery, the neck pain scores of COT group and ET-UA group were (2.13±1.07) and (2.31±1.07) ( P=0.368) ; the neck injury index was (5.13±3.02) and (5.31±3.00) ( P=0.748) ; the dysphagia index was 5 (0.49±3.47) and (3.77±2.96) ( P=0.006) . At 6 months postoperatively, the neck pain scores in COT group and ET-UA group group were (0.52±0.61) and (0.60±0.74) ( P=0.510) ; the neck injury index was (2.10±1.71) and (2.38±2.35) ( P=0.467) ; the dysphagia index was (2.68±2.36) and (1.81±1.83) ( P=0.034) , respectively. Conclusion:Our preliminary study shows that there is no significant differences in postoperative neck pain or discomfort between COT groups and ET-UA group, whereas the impact of ET-UA on swallow function is relatively small than COT.

Objective:To investigate the efficacy, safety and advantages of gasless unilateral axillary approach (GUAA) in endoscopic thyroid surgery.Methods:A total of 334 patients who underwent the GUAA endoscopic thyroid surgery (GUAA group) or conventional open thyroid surgery (OS group) in the Department of Head and Neck Surgery of Zhejiang Cancer Hospital from January 2017 to June 2018 were retrospectively analyzed. There were 45 males and 289 females, aged from 12 to 72 years old, of whom 139 patients were assigned to GUAA group and 195 patients to OS group. Pathological results included papillary thyroid carcinoma (282 cases), nodular goiter (41 cases) and thyroid adenoma (11 cases). Surgical exploration development curve of GUAA group was drawn and was divided into two parts: the technical exploration stage and the technical stable stage. Surgical efficiency, incidences of complications, and incision satisfaction were compared between GUAA group in technical stable stage and OS group. SPSS 25.0 software was adopted for statistical analysis.Results:The mean age in GUAA group was younger than that in OS group, with a significant difference [(35.3±9.5) years vs. (48.1±10.6) years, t=11.31, P<0.01]. The cases in the endoscope group were divided into technical exploration stage for 51 cases and technical stable stage for 88 cases according to the exploration and development curve. In unilateral radical thyroidectomy and unilateral thyroid lobectomy, the mean operation time [(90.6±18.6) min and (93.5±22.0) min] and postoperative drainage volumes [(121.5±87.6) ml and (155.5±69.1) ml] of GUAA group in the stable stage were more than those of OS group [(61.6±15.6) min and (46.5±8.4) min] and [(93.2±42.3) ml and (78.9±48.7) ml]. The difference was statistically significant ( t=12.28, 7.23, 3.35 and 3.05 respectively, all P<0.05), but there were no significant differences in surgical bleeding volumes between two groups [(12.7±6.8) ml vs. (13.5±7.7) ml, t=0.74, P>0.05 and (16.3±14.1) ml vs. (11.9±5.1) ml, t=1.05, P>0.05]. Compared with OS group, GUAA group had the lower incidence of anterior cervical discomfort during swallowing (2.3% vs. 29.2%, P<0.01) and the higher incision satisfaction score (1.1±0.5 vs. 2.8±0.7, t=21.12, P<0.01), however, GUAA group had the higher incidence of supraclavicular (or infraclavicular) numbness after surgery (5.7% vs. 0, P<0.01). And there was no significant difference in the incidences of temporary recurrent laryngeal nerve injury, bleeding, hematoma, infection, lymphatic leakage or chylous leakage after surgery between two groups ( P>0.05). Conclusion:GUAA endoscopic thyroid surgery is a safe method with high cosmetic satisfaction.

Objective: To discuss the relationship between lingual tonsil hypertrophy and laryngopharyngeal reflux. Methods: Ninety-two patients who received throat surgery in Nanfang Hospital between October 2015 and October 2016 were enrolled. Twenty-six healthy volunteers were recruited as normal controls. All participants were assessed with the reflux finding score(RFS) and the size of lingual tonsils were evaluated using a clinical grading system proposed by Friedman under electronic laryngoscope. The score of reflux symptom index(RSI), personal history and medical history were gathered. Biopsy specimens of lingual tonsils were taken from all participants for the immunohistochemical stain of pepsin.SPSS 19.0 software was used for statistical analysis. Results: There were 46.2% (12/26) pepsin-positive and 53.8% (14/26) pepsin-negative volunteers in normal controls. There were 87.0% (80/92) pepsin-positive and 13.0% (12/92) pepsin-negative patients in study group. The severity of lingual tonsil hypertrophy and expression intensity of pepsin in patients were significantly higher in volunteers (Z=-3.636, Z=-5.273, P<0.01). The severity of lingual tonsil hypertrophy was positively associated with the pepsin level in patients (r=0.556, P<0.01). The patients with pepsin-positive expression showed significant correlation between lingual tonsil hypertrophy and the positive rate of RSI and RFS (r=0.258, r=0.225, P<0.05). Analysis of correlated factors indicated that lingual tonsil hypertrophy was associated with smoking (χ²=8.502, P<0.05). Conclusions The expression of pepsin can be detected in lingual tonsil tissues. The lingual tonsil hypertrophy is closely related to laryngopharyngeal reflux.