Objective:To study the effects of different calcium ion concentrations on epithelial mesenchymal transformation (EMT) of human peritoneal mesothelial cell (HPMC) via endoplasmic reticulum stress (ERS).Methods:HPMC cell line HMrSV5 was cultured in vitro and treated in groups. The cells in the control group, high calcium group 1, and high calcium group 2 were treated with medium containing calcium ion concentrations of 1.25, 1.75, and 2.25 mmol/L, respectively. The solvent control group was treated with medium containing 1.25 mmol/L physiological calcium ion concentration and 0.1% dimethyl sulfoxide (DMSO), the high calcium+solvent group was treated with medium containing 2.25 mmol/L calcium ion concentration and 0.1% DMSO, the high calcium+4-phenylbutyric acid (4-PBA) group was treated with medium containing 2.25 mmol/L calcium ion concentration and 1 mmol/L ERS inhibitor 4-PBA, and each group was treated for 48 hours. Morphological changes of cells in each group were observed under light microscope. The expressions of epithelial cell phenotype marker zonula occluden-1 (ZO-1) and mesenchymal cell phenotype marker α-smooth muscle actin (α-SMA) in the cells were observed by immunofluorescence staining. The expressions of EMT marker genes E-cadherin, ZO-1, α-SMA and Vimentin were detected by fluorescence quantitative polymerase chain reaction (PCR). The expressions of ERS marker proteins phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), transcription activating factor 4 (ATF4) and C/EBP homologous protein (CHOP) were detected by Western blotting. Results:Compared with the control group, the morphology of HMrSV5 cells became slender and fibrotic, the fluorescence intensity of ZO-1 increased, and the fluorescence intensity of α-SMA decreased in high calcium 1 and high calcium 2 groups, indicating that the cells transformed from epithelial cells to mesenchyme cells. The mRNA expressions of E-cadherin and ZO-1 were significantly decreased, while the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly increased, moreover, the expressions of the above marker genes or proteins in the high calcium 2 group was more obvious than those in the high calcium 1 group [E-cadherin mRNA (2 -ΔΔCt): 0.53±0.05 vs. 0.75±0.09, ZO-1 mRNA (2 -ΔΔCt): 0.42±0.06 vs. 0.69±0.06, α-SMA mRNA (2 -ΔΔCt): 1.81±0.16 vs. 1.32±0.14, Vimentin mRNA (2 -ΔΔCt): 2.05±0.22 vs. 1.48±0.16, p-PERK protein (p-PERK/β-actin): 0.81±0.09 vs. 0.59±0.06, p-eIF2α protein (p-eIF2α/β-actin): 0.87±0.10 vs. 0.50±0.06, ATF4 protein (ATF4/β-actin): 0.93±0.10 vs. 0.72±0.06, CHOP protein (CHOP/β-actin): 0.79±0.09 vs. 0.46±0.04, all P < 0.05]. Compared with the solvent control group, the morphological changes of cells, the expressions of EMT marker genes and ERS marker proteins after high calcium ion concentration of 2.25 mmol/L were consistent with those in the high calcium 2 group than control group. Compared with the high calcium+solvent group, the cell morphology recovered the characteristics of polygonal and pebble-like epithelial cells in the high calcium+4-PBA group, the fluorescence intensity of ZO-1 increased, the fluorescence intensity of α-SMA decreased, and the mRNA expressions of E-cadherin and ZO-1 in the cells were significantly increased [E-cadherin mRNA (2 -ΔΔCt): 0.86±0.09 vs. 0.57±0.04, ZO-1 mRNA (2 -ΔΔCt): 0.81±0.06 vs. 0.48±0.05, both P < 0.05], the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly decreased [α-SMA mRNA (2 -ΔΔCt): 1.21±0.13 vs. 1.77±0.15, Vimentin mRNA (2 -ΔΔCt): 1.30±0.14 vs. 1.94±0.20, p-PERK protein (p-PERK/β-actin): 0.38±0.04 vs. 0.92±0.11, p-eIF2α protein (p-eIF2α/β-actin): 0.34±0.05 vs. 1.05±0.13, ATF4 protein (ATF4/β-actin): 0.57±0.06 vs. 0.97±0.11, CHOP protein (CHOP/β-actin): 0.51±0.04 vs. 0.90±0.12, all P < 0.05]. Conclusion:High calcium ion concentrations of 1.75 mmol/L and 2.25 mmol/L promote EMT of HPMC via activating ERS.

Objective To understand the current status of research on lung cancer immunotherapy to provide a reference for further investigation and future topic selection in this field. Methods CiteSpace visualization analysis software was used to analyze 400 Chinese studies in CNKI and 5 001 English studies in the Web of Science database from 2005 to 2021, with "lung cancer" and "immunotherapy" as keywords. Keyword co-occurrence analysis was performed on 17 English studies of "Lung Cancer" "Immunotherapy" and "Single cell sequencing" in the Web of Science database. Results "Non-small cell lung cancer" "immunosuppressants" "PD-L1" "dendritic cells" and "cytokine-induced killer cells" are current research hotspots in lung cancer immunotherapy. Monoclonal antibody drugs including nivolumab, pembrolizumab, atezolizumab, and durvalumab are hotspot drugs. Immunotherapy combined with chemotherapy as well as PD-L1 expression have become the focus of continuous research. The majority of studies on lung cancer immunotherapy are conducted in the United States, followed by China. Conclusion Lung cancer immunotherapy has gradually become a research hot spot in China. In the future, in-depth research is needed to provide cutting-edge directions for lung cancer immunotherapy.

Objective:To preliminarily investigate the effect of blue light on the biological activity of human skin keratinocytes, fibroblasts and melanocytes.Methods:Discarded foreskin tissues were collected from 10 healthy children aged from 3 to 12 years after circumcision surgery in the First Affiliated Hospital of Kunming Medical University from June 2021 to December 2021. After epidermis-dermis separation, selective culture was performed to isolate keratinocytes, fibroblasts, and melanocytes. According to the pre-experiment results, the above three types of cells were irradiated with 440 - 450 nm blue light at doses of 0, 5, 10, 20, 30, and 40 J/cm 2, and then continued to be cultured for 0, 6, 24, and 48 hours. Cell counting kit 8 (CCK8) assay was performed to evaluate cellular proliferative activity at each time point, enzyme-linked immunosorbent assay (ELISA) to detect levels of interleukin (IL) -18, IL-33, nerve growth factor (NGF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) secreted by keratinocytes, as well as levels of IL-33 and keratinocyte growth factor (KGF) secreted by fibroblasts, NaOH lysis method to determine melanin synthesis rates in melanocytes, and Western blot analysis to determine the relative expression of tyrosinase (TYR), tyrosine-related protease 1 (TRP-1) and dopachrome isomerase (DCT) in melanocytes. Two-way analysis of variance was used to analyze group effects, time effects and interaction effects. Results:After irradiation with blue light, the cellular proliferative activity significantly differed among different doses of blue light irradiation groups and different time points in keratinocytes ( Ftime = 516.20, Fdose = 421.20, Finteraction = 25.05, all P < 0.003), fibroblasts ( Ftime = 129.30, Fdose = 477.80, Finteraction = 10.91, all P < 0.003), and melanocytes ( Ftime = 77.61, Fdose = 138.70, Finteraction = 3.50, all P < 0.003) ; immediately after irradiation, the proliferative activity of keratinocytes and fibroblasts was significantly lower in the 20 - 40 J/cm 2 blue light group than in the 0 J/cm 2 blue light group (all P < 0.003), and the proliferative activity of melanocytes was significantly higher in the 5 J/cm 2 blue light group than in the 0 J/cm 2 blue light group ( P < 0.003) ; the proliferative activity of the 3 types of cells showed decreasing trends with the increase of blue light irradiation doses and culture time. ELISA showed that the concentrations of IL-18, IL-33, NGF, and GM-CSF secreted by keratinocytes, as well as the concentrations of IL-33 and KGF secreted by fibroblasts, tended to increase with the increase of blue light irradiation doses and culture time. The melanin synthesis rates in melanocytes significantly differed among different doses of blue light irradiation groups and different time points ( Ftime = 833.50, Fdose = 249.40, Finteraction = 81.38, all P < 0.003) ; during 0 - 24 hours after blue light irradiation, the melanin synthesis rates tended to increase with the increase of blue light irradiation doses and time; during 24 - 48 hours, the melanin synthesis rates showed decreasing trends with the increase of blue light irradiation doses and culture time compared with that at 24 hours after irradiation; 24 hours after irradiation, the melanin synthesis rates were significantly higher in the 5, 10, 20, 30 and 40 J/cm 2 blue light groups (159.50% ± 10.88%, 218.76% ± 8.49%, 333.72% ± 7.72%, 393.29% ± 6.00%, 427.21% ± 8.39%, respectively) than in the 0 J/cm 2 blue light group (102.29% ± 6.57%, all P < 0.003). The relative expression of TYR ( Ftime = 67.94, Fdose = 28.99, Finteraction = 3.71, all P < 0.003), TRP-1 ( Ftime = 21.73, Fdose = 8.38, both P < 0.003) and DCT ( Ftime = 34.51, Fdose = 11.79, both P < 0.003) in melanocytes significantly differed among different doses of blue light irradiation groups and different time points, and tended to increase with the increase of blue light irradiation doses and culture time. Conclusion:Blue light irradiation at doses of 5 - 40 J/cm 2 could inhibit the proliferative activity of human skin keratinocytes, fibroblasts, and melanocytes, and the inhibitory effect tended to increase with the increase of blue light irradiation doses, except an enhancing effect on the proliferative activity of melanocytes observed immediately after irradiation with blue light at 5 J/cm 2; additionally, blue light irradiation at 5 - 40 J/cm 2 could enhance the expression of melanin synthesis-related enzymes in melanocytes, and increase the melanin synthesis rate in melanocytes over a short period of time.

ObjectiveTo investigate effect of conducting training of autism spectrum disorder （ASD） early screening skill on improving the ability to early identify ASD of medical staffs in primary care hospitals. MethodsIn September 2021， the training of ASD early screening skills was carried out for medical staffs from 20 primary care hospitals in Chengdu. After training， the training effect was evaluated. The numbers of referrals from primary care hospitals to superior hospitals， confirmed ASD as well as their average diagnostic age of children with ASD before and after training were used as evaluation indicators. ResultsAfter training， the number of children with suspected ASD referred by primary care hospitals was more than that before training ［（16.65±11.60） vs. （3.40±2.23）， t=5.431， P<0.01］， the number of children diagnosed with ASD was more than that before training［（6.85±4.93） vs. （2.45±1.67）， t=4.171， P<0.01］， and the differences were statistically significant. As for the diagnosed age of ASD children， after training， the average age was lower than that before training ［（34.95±11.67） vs. （42.2±14.64）， t=-2.553， P=0.019］. ConclusionTraining of ASD early screening skills for medical staffs in primary care hospitals may help to improve their ability to early screening ASD children.

Objective:To investigate the clinical effect of auricle reconstruction using an expanded postauricular flap with double pedicle in the ear reconstruction.Methods:From September 2016 to August 2017, the clinical data of all patients with congenital microtia treated in the Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The surgical procedures: The expander was implanted in the first stage. The ear framework was covered by the retroauricular double-pedicled expanded flap with free skin grafting at the retroauricular superior margin in the second stage, and the helix was wrapped by the retroauricular fascia. The patients were followed up at 1, 6 months after operation and before the third stage operation, and the appearance of the reconstructed auricle was evaluated by two junior doctors before the third stage of the operations. If there were any differences, the final evaluation would be carried out by a senior doctor. The evaluation results were good, medium and poor.Results:A total of 46 patients (49 ears) with congenital microtia were included, including 36 males (39 ears) and 10 females (10 ears), with the age range of 6-23 years. After operation, the blood supply was good, and the skin grafts survived completely. Three ears had postoperative hematomas under the double-pedicled expanded skin flap when the drainage was removed 5 days after operation. The hematomas recovered well after the negative pressure suction and dressing changed. During the follow-up of 6 months and 18 months, 43 cases (46 ears) were evaluated well with symmetrical and clear three-dimensional structure of reconstructed auricles, uniformed color, and hidden scar behind the ear. Three cases (3 ears) were evaluated as medium. There was hematoma when the drainage tube was removed 5 days after operation, which survived completely after negative suction and dressing change. No cartilage framework infection and exposure were found.Conclusions:The double-pedicled expanded flap has many advantages, such as reliable blood supply, natural auricle appearance, concealed scar, and less skin color difference. It is a good choice for soft tissue wrapping and covering during auricle reconstruction.

Objective:To investigate the clinical effect of auricle reconstruction using an expanded postauricular flap with double pedicle in the ear reconstruction.Methods:From September 2016 to August 2017, the clinical data of all patients with congenital microtia treated in the Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The surgical procedures: The expander was implanted in the first stage. The ear framework was covered by the retroauricular double-pedicled expanded flap with free skin grafting at the retroauricular superior margin in the second stage, and the helix was wrapped by the retroauricular fascia. The patients were followed up at 1, 6 months after operation and before the third stage operation, and the appearance of the reconstructed auricle was evaluated by two junior doctors before the third stage of the operations. If there were any differences, the final evaluation would be carried out by a senior doctor. The evaluation results were good, medium and poor.Results:A total of 46 patients (49 ears) with congenital microtia were included, including 36 males (39 ears) and 10 females (10 ears), with the age range of 6-23 years. After operation, the blood supply was good, and the skin grafts survived completely. Three ears had postoperative hematomas under the double-pedicled expanded skin flap when the drainage was removed 5 days after operation. The hematomas recovered well after the negative pressure suction and dressing changed. During the follow-up of 6 months and 18 months, 43 cases (46 ears) were evaluated well with symmetrical and clear three-dimensional structure of reconstructed auricles, uniformed color, and hidden scar behind the ear. Three cases (3 ears) were evaluated as medium. There was hematoma when the drainage tube was removed 5 days after operation, which survived completely after negative suction and dressing change. No cartilage framework infection and exposure were found.Conclusions:The double-pedicled expanded flap has many advantages, such as reliable blood supply, natural auricle appearance, concealed scar, and less skin color difference. It is a good choice for soft tissue wrapping and covering during auricle reconstruction.

Objective:To explore the research progress and hotspots of nurse-led care models in China, in order to provide references for further research.Methods:The Papers related to nurse-led care models included before 2019 were retrieved from Wanfang database and based on core nursing periodicals from Chinese S&T Journal Citation Reports(Natural Science)(2018 edition), used Bicomb2.0 for word frequency analysis of key words, then used SPSS22.0 for clustering analysis.Results:Totally 118 references and 33 high frequency keywords were retrieved. The number of literature about nurse-led showed a fluctuating upward trend. Researchers could get most of the information about nurse-led intervention mode from Journal of Nursing Science, Chinese Nursing Research and Chinese Nursing Management. By cluster analysis, the hotspots of nurse-led mainly involved critical illness, tumor, chronic disease, community rehabilitation and so on. Conclusions:The nurse-led model has been used in many kinds of diseases and has formed a certain scale, but there is still more room for development. Nursing experts from different regions should strengthen cooperation to improve the nurse-led intervention mode.

Objective:To address a new classification method in term of the three-dimensional space of orbit and to present the different surgical approaches correspondingly.Methods:In a retrospect study from April of 2015 to June of 2018, 102 patients were performed lower eyelid blepharoplasty, including 20 males and 82 females, aged 21-65 years, with an average of 45.2 years. These patients were divided into five groups, which were described in term of three-dimensional structure of orbit, based on the following points: the presence and extent of herniated orbital fat, the presence of inferior orbitopalpebral sulcus, amount of excess skin, and the skin wrinkles in the lower eyelid. And then patients in different group were treated with different kinds of blepharoplasty. All patients in this study ranged in follow-up from 1 month to 12 months. With patients＇ permit, photographs and clinical information were taken to evaluate the preoperative and postoperative outcome.Results:In type 1, all the 32 cases healed well, no complications such as hematoma, infection and ectropion occurred. During the follow-up of more than 1 months, the overall effect was good, and the pouch-shaped appearance of lower eyelid pouch was significantly improved. In types 2, 8 cases had no complications, and the incision healed well; the patients were followed up for more than 3 months, the lower eyelid bag and lower eyelid skin relaxation were significantly improved, and the lower eyelid skin was tighter than before. In types 3, there were no complications in these 19 cases, and the incision was healed well. The patients were followed up for more than 3 months, the deformity of lower eyelid bag was improved obviously, and no obvious local bulge was found under static and dynamic expression. In type 4, there were no complications in all 34 cases, and the incision healed well; during the follow-up of more than 3 months, the lower eyelid pouch deformity and lower eyelid skin relaxation were greatly improved, except for 1 case with mild bulge (untreated). In type 5, 9 cases had no complications, and the incision healed well; during the follow-up of more than 3 months, the fold of the lower eyelid skin disappeared.Conclusions:Little information is available about classification of lower eyelid bags. And the exact surgical approach remains controversial and largely dependent upon surgeon preference and a patient＇s stated cosmetic desire. In this study, an objective classification based on clinical appearance combined with forming reasons of lower eye bags is little available, and the appropriate surgical approach remains controversial as well.

Objective:To investigate the effect of oxygen concentration on proliferation, apoptosis and migration of rabbit auricular chondrocytes at different time points of culture.Methods:Bilateral auricular cartilage from 6 Japanese white rabbits were harvested and cultured, auricular chondrocytes at passage 2 (P2) were used in this study. Five groups were set: Group A (control group): chondrocytes were cultured in atmospheric (21%) oxygen condition, group B: chondrocytes were cultured in 5% oxygen condition for 12 hours (h), group C: chondrocytes were cultured in 5% oxygen condition for 36 h, group D: chondrocytes were cultured in 1% oxygen condition for 12 h, and group E: chondrocytes were cultured in 1% oxygen condition for 36 h. Cell counting kit-8 (CCK-8) assay was adopted to evaluate the proliferation of chondrocytes. Proliferation curves were drawn according to the absorbance value detected. Cell apoptosis was investigated by annexin V-FITC and flow cytometry. Cell scratch test was used to observe the migration ability. One-way ANOVA and LSD- t were used to verify differences between the groups. P<0.05 was considered as statistically significant. Results:In terms of cell proliferation, on the sixth day of culture, the absorbance values of group A, B, C, D and E were 1.38 ± 0.29, 1.59 ± 0.27, 1.37 ± 0.22, 2.06 ± 0.64, 2.23 ± 0.56 respectively. Significant difference was found between the five groups ( F=4.207, P=0.012), and there were significant differences between group D and A ( t=-2.487, P=0.022), group E and A ( t=-3.095, P=0.006). On the seventh day of culture, the absorbance values of the five groups were 1.72 ± 0.30, 2.26 ± 0.44, 2.30 ± 0.29, 2.49 ± 0.73, 2.74 ± 0.54. Significant difference was found between five groups ( F=2.948, P=0.046), and there were significant differences between group D and A ( t=-2.480, P=0.022), group E and A ( t=-3.287, P=0.004). The apoptosis rate of group A, B, C, D and E were (2.97±1.14)%, (3.92±1.14)%, (3.38±0.83)%, (1.54±0.50)%, (0.99±0.59)%. The difference between groups for cell apoptosis were also significant ( F=7.957, P=0.001), among which there were significant differences between group D and A ( t=-2.300, P=0.036), group E and A ( t=-3.180, P=0.006), group B and D ( t=3.812, P=0.002), group C and E ( t=3.832, P=0.002). As for cell migration, at the 12th hour, the cell migration rates of groups A, B, C, D and E were(13.10±3.32)%, (9.23±3.56)%, (10.60±2.03)%, (13.33±1.72)%, (15.32±4.72)%. Significant difference was found between the five groups ( F=3.278, P=0.027). And there were significant differences between group D and B ( t=2.183, P=0.039), group C and E ( t=-2.513, P=0.019). At 24 h, the cell migration rates of the five groups were(19.7±2.97)%, (16.62±3.30)%, (18.99±2.61)%, (20.92±5.18)%, (25.29±5.83)%. Significant difference was found between five groups ( F=3.513, P=0.021). And there were significant differences between group E and A ( t=2.315, P=0.029), group E and C ( t=2.609, P=0.015). Conclusions:When cell culture time exceeded 12 h, 1% oxygen condition induced higher proliferation, lower apoptosis, and higher migration ability than 5% or 21% oxygen conditions in cultured chondrocytes. Moreover, oxygen concentration had stronger influence on the biological characteristics of rabbit auricular chondrocytes compared to intervention time.

Objective:To investigate the effect of oxygen concentration on proliferation, apoptosis and migration of rabbit auricular chondrocytes at different time points of culture.Methods:Bilateral auricular cartilage from 6 Japanese white rabbits were harvested and cultured, auricular chondrocytes at passage 2 (P2) were used in this study. Five groups were set: Group A (control group): chondrocytes were cultured in atmospheric (21%) oxygen condition, group B: chondrocytes were cultured in 5% oxygen condition for 12 hours (h), group C: chondrocytes were cultured in 5% oxygen condition for 36 h, group D: chondrocytes were cultured in 1% oxygen condition for 12 h, and group E: chondrocytes were cultured in 1% oxygen condition for 36 h. Cell counting kit-8 (CCK-8) assay was adopted to evaluate the proliferation of chondrocytes. Proliferation curves were drawn according to the absorbance value detected. Cell apoptosis was investigated by annexin V-FITC and flow cytometry. Cell scratch test was used to observe the migration ability. One-way ANOVA and LSD- t were used to verify differences between the groups. P<0.05 was considered as statistically significant. Results:In terms of cell proliferation, on the sixth day of culture, the absorbance values of group A, B, C, D and E were 1.38 ± 0.29, 1.59 ± 0.27, 1.37 ± 0.22, 2.06 ± 0.64, 2.23 ± 0.56 respectively. Significant difference was found between the five groups ( F=4.207, P=0.012), and there were significant differences between group D and A ( t=-2.487, P=0.022), group E and A ( t=-3.095, P=0.006). On the seventh day of culture, the absorbance values of the five groups were 1.72 ± 0.30, 2.26 ± 0.44, 2.30 ± 0.29, 2.49 ± 0.73, 2.74 ± 0.54. Significant difference was found between five groups ( F=2.948, P=0.046), and there were significant differences between group D and A ( t=-2.480, P=0.022), group E and A ( t=-3.287, P=0.004). The apoptosis rate of group A, B, C, D and E were (2.97±1.14)%, (3.92±1.14)%, (3.38±0.83)%, (1.54±0.50)%, (0.99±0.59)%. The difference between groups for cell apoptosis were also significant ( F=7.957, P=0.001), among which there were significant differences between group D and A ( t=-2.300, P=0.036), group E and A ( t=-3.180, P=0.006), group B and D ( t=3.812, P=0.002), group C and E ( t=3.832, P=0.002). As for cell migration, at the 12th hour, the cell migration rates of groups A, B, C, D and E were(13.10±3.32)%, (9.23±3.56)%, (10.60±2.03)%, (13.33±1.72)%, (15.32±4.72)%. Significant difference was found between the five groups ( F=3.278, P=0.027). And there were significant differences between group D and B ( t=2.183, P=0.039), group C and E ( t=-2.513, P=0.019). At 24 h, the cell migration rates of the five groups were(19.7±2.97)%, (16.62±3.30)%, (18.99±2.61)%, (20.92±5.18)%, (25.29±5.83)%. Significant difference was found between five groups ( F=3.513, P=0.021). And there were significant differences between group E and A ( t=2.315, P=0.029), group E and C ( t=2.609, P=0.015). Conclusions:When cell culture time exceeded 12 h, 1% oxygen condition induced higher proliferation, lower apoptosis, and higher migration ability than 5% or 21% oxygen conditions in cultured chondrocytes. Moreover, oxygen concentration had stronger influence on the biological characteristics of rabbit auricular chondrocytes compared to intervention time.