Objective: To explore the longitudinal changes and related predictive factors of anxiety and intolerance of uncertainty (IU) among Chinese college students, so as to provide theoretical reference for promoting students mental health. Methods: Intolerance of Uncertainty Scale-short Form and the 7 item Generalized Anxiety Disorder Scale were administered among 5 683 students from 22 universities in Guangdong Province from September 10 to October 17, 2021 (T1, baseline survey), and from March 15 to April 22, 2023 (T2). Using paired sample t-test to analyze the changes in anxiety and IU among college students, a linear regression model was established to explore factors that can predict anxiety, IU and their changes during T1 and T2 periods. Results: During the T1 period, the average scores of anxiety and IU among college students were (3.26±3.46, 34.88±7.96), while during the T2 period, they were (4.41±4.16, 36.40±8.07). During the T2 period, the levels of anxiety and IU among college students were higher than those during the T1 period ( t=19.59, 13.67, P <0.01). The linear regression results showed that age ( B = -0.02 ), moderate ( B =-0.18) and poor ( B =-0.88) mental health status, as well as online browsing of Corona Virus Disease 2019 (COVID-19) epidemic (later called as epidemic) related information for >1-<3 hours ( B =-0.30) and ≥3 hours ( B =-0.22), all of which could negatively predict changes in anxiety during T1 to T2 periods. Living in rural areas ( B =0.11) could positively predict changes in anxiety during T1 to T2 periods ( P <0.05). Moderate ( B =-0.19) and poor ( B =-0.47) mental health status, browsing epidemic related information online for >1-<3 hours ( B =-0.32) and ≥3 hours ( B =-0.33), academic performance being moderately affected by the epidemic ( B =-0.10), and personal planning being moderately affected by the epidemic ( B =-0.13) and severely affected ( B =-0.22), all of which could negatively predict changes in IU during T1 to T2 periods ( P <0.05). Conclusions Levels of anxiety and intolerance of uncertainty increases significantly after epidemic. Mental health status of college students should need long term monitoring, and students with poor mental health should need special attention.

Objective To investigate the regulatory role of ubiquitin-specific protease 10(USP10)on the protein expression of Krüppel-like factor 4(KLF4)and its impact on the proliferation and invasion ability of hepatocellular carcinoma(HCC)cells.Methods The protein expression differences of USP10 and KLF4 in normal liver cell line L02 and HCC cell lines,including HepG2,HUH7,HCCLM3 were detected by immunoblotting(Western blot)methods.HCCLM3 and HUH7 cells were selected,and lentiviral particles overexpressing or silencing USP10(oe-USP10 or sh-USP10)was transfected into the cells,and they were designated as the oe-USP10 group and oe-NC group,respectively.Immunoprecipitation(Co-IP)experiments were conducted to examine whether USP10 could di-rectly interact with KLF4 in HCCLM3 or HUH7 cells.The Co-IP assay was repeated in HCC cells transfected with oe-USP10 or sh-USP10,with the addition of the proteasome inhibitor MG132,which used to detect the ubiquitina-tion level of KLF4 protein in the transfected HCC cells.The pcDNA3.1 vector containing overexpressed KLF4 or its negative control plasmid(pc-KLF4 or pc-NC)was co-transfected into cells of the sh-USP10 group or sh-NC group.These cells were designated as the sh-NC+pc-NC group,sh-USP10+pc-NC group,sh-NC+pc-KLF4 group,and sh-USP10+pc-KLF4 group.The cell proliferation activity of each group was measured using the CCK-8 assay,and the cell invasion ability was assessed using the Transwell assay.Results Compared to L02 cells,the protein expres-sion of USP10 and KLF4 significantly decreased in HepG2,HUH7,HCCLM3,and other cells(P＜0.05).In HC-CLM3 and HUH7 cells,USP10 protein directly interacted with KLF4.Furthermore,treatment with MG132 resulted in a time-dependent increase in KLF4 protein expression in HCCLM3 and HUH7 cells.Silencing USP10 increased the ubiquitination of KLF4 in HCCLM3 or HUH7 cells,while overexpressing USP10 decreased the ubiquitination level of KLF4 in cells.Compared to the sh-NC+pc-NC group,both the proliferation activity and invasion ability of HCCLM3 and HUH7 cells significantly increased in the sh-USP10+pc-NC group(P＜0.01),while they signifi-cantly decreased in the sh-NC+pc-KLF4 group and sh-USP10+pc-KLF4 group(P＜0.05).Compared to the sh-USP10+pc-NC group,the proliferation activity and invasion ability of cells significantly decreased in the sh-USP10+pc-KLF4 group(P＜0.05).Conclusion USP10 can promote the stability of KLF4 protein through deubiquiti-nation in HCC cell lines,thereby inhibiting the proliferation and invasion of tumor cells.

Objective:To investigate whether IL-7-secreting oncolytic herpes simplex virus(HSV)could activate CD8+T cells and inhibit the growth of hepatocellular carcinoma.Methods:The expression of IL-7 was detected by Western blot.The in vitro cleavage of tumor cells by tumor oncolytic virus HSV and HSV-IL-7 were detected by crystal violet staining.The tumor inhibition ability of HSV-IL-7 and HSV were detected in subcutaneous transplanted tumor model.Levels of IL-7,IFN-γ and TNF-α in serum and tumor tissues were determined by ELISA.The infiltration of CD8+T cells in tumor tissues was detected by immunohistochemistry.Flow cytometry was used to detect Granzyme B secretion in CD8+T cells infiltrated by tumor.Results:Tumor cells infected with HSV-IL-7 expressed high level of IL-7.Both HSV and HSV-IL-7 can effectively lyse B16-F10,CT-26 and H22 tumor cell lines in a dose-dependent manner in vitro.HSV-IL-7 could significantly inhibit the growth of H22 hepatoma cells in vivo(P<0.01)and prolong the survival time of tumor-bearing mice(P<0.001).HSV-IL-7 could significantly increase the IL-7 content in tumor sites(P<0.000 1),and effectively increase the number of tumor infiltrating CD8+T cells(P<0.001).HSV-IL-7 significantly enhanced Granzyme B secretion of tumor-infiltrating CD8+T cells and IFN-γ and TNF-α in tumor tissues(P<0.000 1).Conclusion:HSV-IL-7 has well tumor inhibition activity in vivo and in vitro.It also can activate the anti-tumor activity of CD8+T cells in vivo by secreting IL-7,inhibit tumor growth and prolong the survival time of tumor-bearing mice.

Objective:To investigate the prognostic value of the combined preoperative albumin-globulin score (AGS) and skeletal muscle index (SMI), referred to as the CAS classification, in predicting postoperative outcomes in patients with pancreatic cancer.Methods:The clinical data from 265 patients who underwent surgical treatment and were pathologically confirmed to have pancreatic cancer at the Affiliated Hospital of Qingdao University between January 2012 and December 2022 were retrospectively analyzed. Patients were randomly divided into a training group ( n=184) and a validation group ( n=81) in a 7∶3 ratio. Patients' age, gender, body mass index (BMI), smoking history, alcohol consumption history, previous history of metabolic diseases, AGS, SMI, and CAS classifications within 7 days before surgery, preoperative upper abdominal CT imaging features, presence of vascular and neural invasion, and lymph node metastasis were recorded. Patients with AGS grade 0 were classified into the low AGS group ( n=48), while those with AGS grades 1 and 2 were classified into the high AGS group ( n=136). The optimal cutoff value for SMI was determined using X-tile software: male patients with SMI>42.6 cm 2/m 2 or female patients with SMI>37.8 cm 2/m 2 were categorized into the high SMI group ( n=125), while those below these thresholds were categorized into the low SMI group ( n=59). Patients with AGS grade 0 and SMI>42.6 cm 2/m 2 for males or >37.8 cm 2/m 2 for females were classified into the CAS grade 1 group (n=32). Patients with AGS grades 1 or 2 and SMI ≤42.6 cm 2/m 2 for males or ≤37.8 cm 2/m 2 for females were classified into the CAS grade 3 group ( n=43). The remaining patients were classified into the CAS grade 2 group ( n=109). Clinical characteristics were compared across these groups. Cumulative survival rates were estimated using the Kaplan-Meier method, and survival curves were plotted to analyze the relationship between AGS, SMI, and CAS classifications and overall survival after pancreatic cancer surgery. Differences among groups were assessed using the Log-Rank test. Receiver operating characteristic curves (ROC) were plotted, and the area under the curve (AUC) was calculated to evaluate the predictive efficacy of AGS, SMI, and CAS on postoperative survival. Results:Compared to the high AGS group, the low AGS group exhibited higher SMI values [(46.17±9.63) cm 2/m 2vs (44.11±7.43) cm 2/m 2], and a lower incidence of lymph node metastasis (16 vs 66, 33.3% vs 48.5%). The mortality rate in the low AGS group was 50.0%(24/48), significantly lower than the 70.6% (96/136) observed in the high AGS group, with a median overall survival of 22.08 months (95% CI 16.87-29.62) longer than 13.1 months (95% CI 8.84-18.82) in high AGS group. Compared to the low SMI group, the high SMI group had a lower prevalence of metabolic diseases (26.4% vs 44.1%). The mortality rate in the low SMI group was 78.0% (46/59), higher than the 58.4% (73/125) in the high SMI group, with a median overall survival of 12.97 months (95% CI 9.37-18.20) obviously shorter than 16.20 months (95% CI 10.7-24.12) in high SMI group. Lymph node metastasis rate for CAS grade 1, 2, and 3 was 34.4% ( n=11), 44.0% ( n=48), and 62.8% ( n=27), respectively, with corresponding mortality rate of 34.3% (11/32), 67.9% (74/109), and 79.1% (34/43), and median overall survival time of 25.55 months (95% CI 19.49-30.07), 14.10 months (95% CI 10.22-19.14), and 12.5 months (95% CI 8.53-18.00), respectively. All the differences were statistically significant (all P value <0.05). Kaplan-Meier survival analyses demonstrated that patients in the low AGS group had significantly longer overall survival than those in the high AGS group in both the training and validation cohorts. Similarly, patients in the high SMI group had longer overall survival compared to those in the low SMI group. Notably, patients in CAS grade 1 exhibited the longest overall survival, whereas those in CAS grade 3 had the shortest. ROC curve analysis revealed that the AUC for CAS classification was superior in the training cohort (0.649) compared to AGS (0.588) and SMI (0.593), and in the validation cohort (0.644) compared to AGS (0.587) and SMI (0.577). Conclusions:CAS classification could effectively predict postoperative prognosis in pancreatic cancer patients, with higher CAS grades correlating with poorer outcomes.

Objective To construct a survival prediction model for the patients with extramedullary plasmacytoma (EMP),and to evaluate its application value.Methods The data of the patients diagnosed as EMP from 2000 to 2019 were collected from the National Cancer Institute's Surveillance,Epidemiology,and End Results (SEER) database.Random forest algorithm was used to screen variables,multivariate Cox pro-portional hazards regression model was used to determine the independent risk factors of EMP prognosis,and a nomogram model was constructed based on these factors.The total score of the patients was calculated ac-cording to the nomogram model,and the total samples were divided into the low,medium and high risk groups according to the optimal cutoff value by using X-tile software,and the Kaplan-Meier survival curves of the three groups were analyzed.The predictive performance of the model was evaluated using area under the curve (AUC) of time-dependent receiver operating characteristic (t-ROC) curve,AUC after X years N repeated K-fold cross-validation,calibration curve,and decision curve analysis (DCA).Results A total of 1458 patients with EMP were included,including 796 cases of deletions and 662 cases of death.In the deletion and deaths groups,the majority were aged 60-<75 years old (35.4％,41.2％),male (63.1％,66.3％),white race peo-ple (81.3％,80.4％),and married (68.2％,59.4％).Finally,age (45-<60 years old/60-<75 years old/≥ 75 years old),marital status (single/others),non-primary malignant tumor,and without surgery and radio-therapy were determined to be the independent risk factors for the survival of EPM patients (P<0.05).Based on the above independent risk factors,a survival prediction model was constructed,and a nomogram was drawn.According to the optimal cutoff value,the total sample was divided into the score of 0-96 group (low risk group),score of 98-135 group (medium risk group) and score of 139-191 group (high risk group).The Kaplan-Meier survival analysis revealed that there was statistically significant difference in the survival time a-mong the three groups (P<0.0001).AUC of t-ROC curves for 1,3,5 years and AUC after cross-validation all were>0.7,indicating that the model had a good degree of differentiation.The calibration curves suggested a good consistency between the prediction and practical over survival rate,and DCA indicated that the model could improve the clinical benefits.Conclusion The constructed survival prediction model for EMP patients based on the SEER database could help the clinicians to identify the prognostic risk factors and predict the o-verall survival rate of the patients.

Objective To distinguish Khat from its similar plants,such as white tea and Houttuynia cordata,so as to identity Khat and its camouflage and confusing products in smuggling crime and public security anti-drug work.Methods FTIR is used to collect the infrared spectral data of plant samples,and the data are analyzed by chemometrics.The original data were preprocessed by three data processing methods,namely,SG smoothing,baseline and normalization,then the processed data were classified and reduced by PCA,finally the mathematical model was constructed by three methods:KNN,RF and SVM for classification and prediction.Results The results were expressed by confusion matrix,and the accuracy of the RF model is 96.51%.The accuracy of the SVM model is 98.84%;The accuracy of the KNN model is 93.02%.Conclusion In this experiment,FTIR was used to establish a model for rapid identification of Khat grass,the SVM model has the highest correct recognition rate and good parameters,which can be used to distinguish dry khat from its similar plants.

Objective:To investigate the clinical characteristics of de novo malignancies (DNMs) after liver transplantation (LT) and to study the clinical management strategies.Methods:Adult LT recipients who were regularly followed-up in the Organ Transplantation Center, the Affiliated Hospital of Qingdao University from January 2005 to April 2021 were enrolled in this study. The clinical characteristics of DNMs were retrospectively analyzed. Of 601 LT recipients, there were 105 females and 496 males, aged (51.4±9.6) years old. They were divided into the DNMs group ( n=26) and the non-DNMs group ( n=575) according to whether there were DNMs on followed-up. Clinical data including age, sex, basic diseases before LT and operation time were collected. These patients were follow-up in outpatient clinics. Results:Twenty-six patients were diagnosed to develop DNMs after LT, but there were 28 DNMs (of which 2 patients were diagnosed to have DNMs twice). The incidence of DNMs after LT was 4.3% (26/601), the median time from LT to DNMs was 42 (20, 70) months, and the cumulative incidence rates of DNMs were 0.5%, 2.0%, 6.3%, 21.0% and 34.5% at 1, 3, 5, 10 and 15 years after LT, respectively. Among the 28 DNMs, digestive system tumors were most common, with 17 lesions (60.7%), followed by 3 lesions (11.1%) of lung cancer, 2 lesions (7.4%) of lymphoproliferative diseases, and 1 lesion (3.7%) of cervical cancer, thyroid cancer, soft palate cancer, eyelid cancer, laryngeal cancer, and prostate cancer. The follow-up time of 55.9 (36.6, 102.5) months in the DNMs group after LT was longer than the 33.4 (18.5, 58.9) months in the non-DNMs group ( P<0.001). The 1, 5, and 10 year survival rates of patients with DNMs after LT were 96.3%, 83.5%, and 49.8%, respectively. The 1, 5, and 10 year survival rates of patients with non-DNMs after LT were 94.5%, 77.7%, and 75.4%, respectively. There was no significant difference in the cumulative survival rates between the two groups (log rank=0.402, P=0.526). Conclusion:The incidence of DNMs in LT recipients was 4.3%. The majority of them were digestive system tumors. Early diagnosis and treatment of DNMs significantly improved the prognosis and quality of life of these patients.

Objective:To detect serum levels of vitamin A (Vit A), vitamin D(Vit D)25-hydroxy vitamin D[25-(OH)D] and vitamin E(Vit E) in children aged 0-6 years in Tibetan Plateau of Garzi Prefecture, thus providing references for physical examinations and prevention of 4 key diseases (rickets, malnutrition anemia, pneumonia and diarrhea) in children in plateau areas by relevant government departments.Methods:A total of 2 122 children who participated in physical examination in 12 townships of Xiangcheng County and 14 townships of Daocheng County, Garzi Tibetan Autonomous Prefecture, Sichuan Province from April 2017 to April 2019 with 0-6 years old were recruited for surveying physical measurements and collection of venous blood.Serum Vit A and Vit E levels were detected by high performance liquid chromatography.Serum levels of 25-(OH)D were detected by high performance liquid chromatography tandem mass spectrometry.The relationship between Vit A, Vit E and 25-(OH)D levels with the gender, age, seasonal change and altitude was analyzed.Results:The serum Vit A level, subclinical Vit A deficiency rate and marginal vitamin A deficiency rate were(1.05±0.27) μmol/L, 8.15%(173/2 122 cases) and 45.99%(976/2 122 cases), respectively in 2 122 children with 0-6 years old.There were significant differences in the serum Vit A level, the subclinical Vit A deficiency rate and the marginal vitamin A deficiency rate in children with different ages, seasons and altitudes (all P<0.05). The serum level of 25-(OH)D and 25-(OH)D deficiency rate insufficient rate were (24.65±6.45) ng/L, 6.03%(128/2 122 cases) and 16.59%(352/2 122 cases), respectively.There were significant differences in the serum level of 25-(OH)D, 25-(OH)D deficiency rate and 25-(OH)D insufficient rate in children with different ages and seasons (all P<0.05). The mean serum Vit E level, Vit E deficiency rate and Vit E insufficient rate were (7.81±1.74) mg/L, 2.78%(59/2 122 cases) and 29.59%(628/2 122 cases), respectively.There were significant differences in serum Vit E level, Vit E deficiency rate and Vit E insufficient rate in children with different ages and seasons (all P<0.05). The mean serum levels of Vit A and Vit D remained the lowest before the age of 1 year, and their deficiencies at this age were the most significant.The mean serum level of Vit E remained the lowest in >1-2 years old, and its deficiency and insufficient at this age were the most significant.Vit A, D and E levels were significantly affected by seasonal changes, which were significantly higher in the summer than in the spring, autumn and winter.In addition, Vit A and 25-(OH)D were significantly affected by the altitude, which were the lowest above 4 km altitude. Conclusions:The overall serum levels of Vit A, 25-(OH) D and E in children with 0-6 years old in Tibetan Plateau areas of Ganzi Prefecture are lower than those in plain areas.Vit A, 25-(OH) D and Vit E levels significantly differed in the age, season and altitude, which are related to the lack of local resources, insufficient maternal nutrition during pregnancy and insufficient intake after birth, as well as temperature and light caused by changes in local seasons and altitude.Therefore, it is necessary to make reasonable supplements during pregnancy to prevent vitamin deficiency.

Objective To investigate the methods and results of hand reconstruction by transplanting the fostered residual finger and the second toe,which share the same blood supply system.Methods From January,2014 to January,2016,3 cases with destructive hand injuries in our hospital;and debridement was performed in one session during the early stage of trauma,with skin grafting to cover the wrist stump and foster the residual finger to a branch of the dorsalis pedis artery.The first dorsal metatarsal artery around the toe web was identified and then gradually isolated proximally with dissection of the branches of the dorsalis pedis artery supplying the fostered finger until the convergence of the two blood vessels.Then,reconstructed hand by residual finger and combined second toe transplantation.Results After a follow-up period of 3 months to 2 years,the reconstructed hand had a good appearance and was able to complete basic mundane actions.The function of the limb was restored to a large extent andthe patient was satisfied with the reconstructed hand.Conclusion Transfer of a fostered residual finger and combined second toe which have the same blood supply system for hand reconstruction as a new method need to anastomose only one set of arteries,and it is feasible with the good appearance and function of the reconstructed hand.

Objective To investigate the methods of repair the lateral skin and soft tissue defect of the middle and rear foot with the plantar medial flap transferred before the achilles tendon.Methods Fifteen cases with the lateral skin and soft tissue defect of the middle and rear foot were repaired with the plantar medial flap pedicled with posterior tibia artery transferred before the achilles tendon from January,2012 to October,2015.All the 15 patients were followed up in the way by telephone or back to hospital postoperatively 3 months to 3 years,including 6 cases by telephone follow-up and 9 cases of back-to-hospital follow-up,to observe the appearance,functions of the skin flap and whether the skin flap survived well.Results The flaps of 15 cases survived well.Postoperative followup of 3 months to 3 years,the sensation of the flap was good,without any ulcer or necrosis.The patients walked well and were satisfied with the flap.Conclusion To repair the lateral skin and soft tissue defect with the plantar medial flap transferred before the achilles tendon as a new method is feasible with the good ability of faster sensory recovery,the good appearance and function.