Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

AIM: To investigate the preoperative ocular symptoms and the characteristics of asymptomatic ocular surface abnormalities in hospitalized patients with primary pterygium.METHODS: Cross-sectional study. Hospitalized patients diagnosed with primary pterygium and scheduled to receive pterygium excision surgery at the Xiamen Eye Center of Xiamen University from August 2022 to October 2022 were enrolled. Ocular surface disease index questionnaire(OSDI), six examinations including non-invasive tear film break-up time, Schirmer I test, tear meniscus height, lid margin abnormality, meibomian gland dropout and tear film lipid layer thickness, and anterior segment optical coherence tomography(AS-OCT)were performed and statistically analyzed.RESULTS: A total of 178 cases(178 eyes), with a mean age of 54.39±10.75 years old, were recruited, including 75 males(42.1%)and 103 females(57.9%). The average values of ocular surface parameters in these patients included OSDI: 11.47±9.69, tear film break-up time: 7.10±3.86 s; tear meniscus height: 0.16±0.07 mm, Schirmer I test values: 14.39±7.29 mm/5 min, and pterygium thickness: 504.74±175.87 μm. Totally 161 eyes(90.4%)presented with abnormal lid margin, 44 eyes(24.7%)presented with meibomian gland dropout score ≥4, 52 eyes(29.2%)presented with low lipid layer thickness. In the 6 objective examinations, abnormalities in at least 4 of these tests were found in 85.4% of eyes. Pterygium morphology was classified into four grades: 10 eyes(5.6%)of grade Ⅰ, 93 eyes(52.2%)of grade Ⅱ, 60 eyes(33.7%)of grade Ⅲ, and 15 eyes(8.4%)of grade Ⅳ. In patients with a higher grade of pterygium, the tear film break-up time was lower, and the proportion of abnormal lid margin was also significantly higher(P<0.05). The patients were further divided into two subgroups, including 121 eyes(68.0%)with normal OSDI <13 in the normal group and 57 eyes(32.0%)with OSDI ≥13 in the abnormal group. No significant difference was found in the proportion of meibomian gland dysfunction between the two groups of patients(71.9% vs. 71.9%, P=0.872). In addition, there were differences in the number of abnormal objective examinations(4.11±0.85 vs. 4.91±0.99, P<0.001).CONCLUSIONS: Asymptomatic ocular surface abnormalities were present preoperatively in patients hospitalized for primary pterygium. A comparable high incidence of structural or functional meibomian gland dysfunction existed in pterygium patients with or without apparent ocular discomfort. More attention should be paid to the ocular surface abnormalities in those asymptomatic patients before primary pterygium surgery.

This paper briefly introduces the definition, classification and significance of effect modification in epidemiological studies, summarizes the difference between effect modifier and confounders, and analyze the influence as well as the role of effect modification in epidemiological studies and Meta-analysis. In this paper, the possible scenarios of effect modification and related analysis strategy in Meta-analysis are indicated by graphics, aiming to arouse researchers' attention to effect modification. This paper also demonstrates how to identify and deal with effect modification in Meta-analysis through a study case of "Efficacy of sodium-glucose cotransporter 2 inhibitors in patients with type 2 diabetes", and shows the analysis process and interpretation of results of subgroup analysis and Meta-regression methods respectively. The advantages and disadvantages of these two methods are summarized to provide reference for the method selection of future research.

This paper briefly introduces the characteristics, research significance, and global reporting status of effect modification in network Meta-analysis, demonstrates the heterogeneity caused by effect modification in network Meta-analysis, and emphasizes the importance of exploring effect modification in network Meta-analysis. This paper also summarizes the normalized description and analysis strategies of effect modification in network Meta-analysis. Finally, by the case of "comparison of efficacy of three new hypoglycemic drugs in reducing body weight in type 2 diabetes patients", this paper demonstrates the realization of subgroup analysis and network Meta-regression in exploring effect modification, summarizes the advantages and disadvantages of the two methods, to provide references for future researchers.

This paper briefly introduces the unique advantages, overall analysis ideas and existing analysis methods of individual patient data Meta-analysis in terms of effect modification. In addition to Meta-regression and subgroup analysis, this paper also introduces the analysis methods based on part of individual patient data integrated with aggregated data and summarizes the current reporting of the above mentioned methods. In addition, the application and results interpretation of the above mentioned methods in individual patient data Meta-analysis are presented in this paper by taking "Effects of sodium-glucose cotransporter 2 inhibitors on SBP in patients with type 2 diabetes" as an example and by introducing their advantages and limitations.

Objective:To investigate the correlation between pen-holding posture, grip strength and myopia.Methods:A cross-sectional study was performed.A total of 496 school-aged children in grades 1-6, who underwent eye examinations at Shenzhen Eye Hospital from November 2021 to May 2022 were consecutively enrolled as subjects.The subjects' age, grade, sex, refractive error and age of school enrollment were recorded.Subjects' grip strength and pen-holding posture were obtained with an electronic grip strength meter and a tool pen, and subjects were divided into correct pen-holding posture group and incorrect pen-holding posture group according to their pen-holding posture.Propensity scores for sex, grade, age at school entry, and grip strength in the two groups were matched using the nearest neighbor matching method.Multifactorial binary logistic regression analysis was performed with binocular myopia, monocular myopia, and emmetropia as dependent variables and grip strength, grade, refractive error, and pen-holding posture as independent variables.The correlation between grip strength and spherical equivalent was examined by Spearman rank correlation analysis and partial correlation analysis.This study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Shenzhen Eye Hospital (No.2022KYPJ032).Results:After matching propensity scores, there were 101 cases and 197 cases in the correct pen-holding posture group and incorrect pen-holding posture group, respectively.There was no statistical significance in sex, grade, age at enrollment, and grip strength between the two groups (all at P>0.05).Multinomial logistic regression results showed that using emmetropia as a reference, higher grade (4 versus 1∶7.601[1.307-44.206]; 5 versus 1∶4.392[1.039-18.562]), presence of anisometropia (21.366[5.750-79.397]) were relative risk factors for monocular myopia (all at P<0.05), and higher grade (3 versus 1∶4.389[1.783-10.800]); 4 versus 1∶15.398[3.267-72.574]; 5 versus 1∶7.447[2.232-24.851]; 6 versus 1∶6.462[2.116-19.734]) were relative risk factors for binocular myopia (all at P<0.05).Taking monocular myopia as a reference, higher grade (6 versus 1∶4.582[1.193-17.589]), presence of anisometropia (0.141[0.069-0.289]), and irregular pen-holding posture (2.608[1.340-5.075]) were associated factors for binocular myopia (all at P<0.05).According to partial correlation analysis, the spherical equivalent of the right eye of children with incorrect pen-holding posture was negatively correlated with the grip strength ( rs=-0.141, P=0.047). Conclusions:There is an association between pen-holding posture and myopia in school-aged children.Children with greater grip strength and incorrect pen-holding posture may have higher degrees of right eye refractive error.

Objective:To evaluate the effectiveness and safety of sofosbuvir/velpatasvir (SOF/VEL) with or without ribavirin in the treatment of patients diagnosed with chronic hepatitis C (CHC) and chronic kidney disease (CKD).Methods:From June 2018 to May 2022, a total of 75 patients with CHC and CKD, and treated with SOF/VEL±ribavirin at the Kunming Third People′s Hospital were enrolled in this study. The basic information of patients were collected. Assessments of renal function, liver function, virologic response rate and adverse events were conducted at baseline, four weeks and 12 weeks of treatment and 12 weeks after treatment withdrawal. Wilcoxon rank sum test and Kruskal-Wallis rank sum test were used for statistical analysis.Results:Among the 75 patients, 51 cases(68.0%) were classified as CKD stage 2, 12 cases (16.0%) as CKD stage 3, four cases (5.3%) as CKD stage 4, eight cases (10.7%) as CKD stage 5. Additionally, 26 cases (34.7%) were classified as HCV type 3a, while 37 cases (49.3%) were classified as type 3b. Among the patients, 51 cases (68.0%) had cirrhosis, including 15(20.0%) compensated cirrhosis and 36(48.0%) decompensated cirrhosis. Twelve weeks after treatment withdrawal, there was a statistically significant improvement in the estimated glomerular filtration rate (eGFR) compared to baseline (81.76(60.94, 94.34) mL/(min·1.73 m 2) vs 70.99(52.86, 82.38) mL/(min·1.73 m 2), Z=8.12, P=0.040). From baseline to 12 weeks after treatment withdrawal, eGFR of patients with CKD stage 2 and stage 3 were both gradually increased, with statistical significance ( H=8.91 and 8.03, respectively, both P<0.05). For CKD stage 2 patients, eGFR increased from 78.82(70.98, 84.80) mL/(min·1.73 m 2) to 86.94 (75.91, 96.01) mL/(min·1.73 m 2), while CKD stage 3 patients had an increased from 51.24 (45.92, 53.37) mL/(min·1.73 m 2) to 64.58 (44.54, 74.34) mL/(min·1.73 m 2). Renal function was improved to CKD stage 1 in 21 patients (28.0%). Compared to baseline, CKD stage 2 patients exhibited a decrease of aspartate aminotransferase to platelet ratio index 12 weeks after treatment withdrawal, and alanine aminotransferase and aspartate aminotransferase levels were also significantly improved with statistical significance ( Z=8.03, 21.57 and 43.74, respectively, all P<0.05). The rate of sustained virological response (SVR)12 at 12 weeks after treatment withdrawal was 98.7%(74/75). Among these cases, 51 patients in CKD stage 2, 11 patients in CKD stage 3, 12 patients in CKD stage 4 and stage 5 reached SVR12. Adverse events occurred in 32 patients (42.7%), including 18 cases of mild hemolytic anemia, four cases of skin itching, three cases of rash, two cases of chest tightness, and five cases of fatigue. Conclusions:SOF/VEL with or without ribavirin for the treatment of patients with CHC and CKD has good effectiveness and safety. The renal function, liver function and liver fibrosis degree have been improved after antiviral treatment.

Objective:To study the genetic profile of neonatal hyperbilirubinemia with unknown etiology in Guangdong Province and the clinical significance of jaundice-related genetic screening.Methods:From July to September, 2021, neonates with hyperbilirubinemia of unknown etiology born in different hospitals in Guangdong Province were studied. 24 neonatal jaundice-related exons were sequenced using targeted capture and high-throughput sequencing technology. The pathogenic variants were analyzed.Results:A total of 331 cases, 139 (42.0%) cases showed positive screening results with five diseases, including 65 (19.6%) cases of Gilbert syndrome, 48 (14.5%) cases of glucose-6-phosphate dehydrogenase (G6PD) deficiency,18 (5.4%) cases of sodium taurocholate cotransporting polypeptide deficiency, 4 (1.2%) cases of Citrin deficiency and 4 (1.2%) cases of Dubin-Johnson syndrome. 149 (45.0%) cases carried one or more genetic variants and 43 (13.0%) cases showed no clinically significant variants. The 8 high-frequency mutation loci (carrier rate >1%) are UGT1A1 gene c.211G>A and c.1091C>T, G6PD gene c.1466G>T and c.1478G>A, SLC10A1 gene c.800C>T, SLC25A13 gene c.852_855del TATG, HBB gene c.126_129delCTTT and c.316-197C>T.Conclusions:Genetic factors are important for neonatal hyperbilirubinemia with unknown etiology in Guangdong. The common pathogenic genes are UGT1A1, G6PD, SLC10A1, and SLC25A13 and the population carries high-frequency mutation loci. Therefore, genetic screening in neonates with hyperbilirubinemia of unknown etiology has important clinical significance.

Objective:To construct the prediction model for the prolonged length of postoperative hospital stay in the patients undergoing thoracoscopic lobectomy.Methods:The patients of both sexes, aged ≥18 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ, who received elective thoracoscopic lobectomy with general anesthesia from March 2016 to February 2019 in our hospital, were selected, their clinical data were collected, and the patients were pathologically diagnosed with non-small-cell lung cancer after operation.Basic information (sex, age, smoking history), previous history (dyslipidemia, hypertension, diabetes, cardiovascular and cerebrovascular diseases, peripheral vascular diseases, chronic obstructive pulmonary diseases), allergy history, other tumor history, surgical resection site, anesthetic factors (intraoperative use of non-steroidal anti-inflammatory drugs and glucocorticoids, duration of anesthesia, intraoperative epidural anesthesia + postoperative epidural analgesia) and postoperative complications (pleural effusion, pneumothorax, atelectasis) was collected.The patients were divided into 2 groups according to whether the length of postoperative hospital stay was prolonged: normal group (≤ 7 days) and prolonged group (>7 days).Logistic regression analysis was used to identify the predictors for prolonged length of postoperative hospital stay.The regression model for prediction of prolonged length of postoperative hospital stay was constructed based on the TensorFlow deep learning framework, and the efficacy of prediction was evaluated.A deep neural network was further established based on the TensorFlow framework to construct a classification prediction model for prolonged length of postoperative hospital stay, and the efficacy of prediction was assessed, further comparing it with the prediction model constructed by the traditional machine learning method.Results:A total of 428 patients were finally enrolled in the study.The results of multivariate logistic regression analysis showed that age and anesthesia duration were the risk factors for the prolonged length of postoperative hospital stay, and female, other tumor history and resection of right middle lobe were the protective factors ( P<0.05).The performance of the regression model proved ineffective, getting 2.16 mean absolute error and 11.05 mean square error on the training set, 2.14 mean absolute error and 11.73 mean square error on the test set.The classification model achieved better score with accuracy 75.58%, F1-measure 0.553 and area under the receiver operating characteristic curve 0.702 on the test set, however, it showed no better performance than that of 4 other prediction models established by 4 traditional machine learning methods, specifically Logistic Regression, Random Forest, Gradient Boosting and Support Vector Machine. Conclusions:Sex, age, surgical resection site, other tumor history and duration of anesthesia can serve as the predictors, and a classification prediction model for prolonged length of postoperative hospital stay is constructed based on a deep neural network in the patients undergoing thoracoscopic lobectomy.

Objective To understand the epidemiological characteristics of the cases of COVID-19 epidemic clusters, and explore the influence of family factors and social factors such as group activities on the spread of the disease. Methods The data of cases of COVID-19 epidemic clusters from 19 January, 2020 to 25 February, 2020 were collected from the official platforms of 36 cities in 6 provinces in China. Descriptive statistical methods, χ 2 test and curve fitting were used to analyze the epidemiological characteristics of the clustered cases. Results By 25 February, 2020, the data of 1 052 cases in 366 epidemic clusters were collected. In these clustered cases, 86.9%(914/1 050) occurred in families. Among the 1 046 cases with gender information, 513 were males (49.0%) and 533 were females (51.0%). The cases were mainly young adults between 18 and 59 years old, accounting for 68.5% (711/1 038). In the 366 epidemic clusters , the clusters in which the first confirmed cases with the history of sojourn in Wuhan or Hubei accounted for 47.0%(172/366). From 19 January to 3 February, 2020, the first confirmed cases with Wuhan or Hubei sojourn history accounted for 66.5%. From 4 to 25 February, the first confirmed cases who had Wuhan or Hubei sojourn history accounted for only 18.2%. The median of interval between the first generation case onset and the second generation case onset was 5 (2-8) days. The median of onset- diagnosis interval of the initial cases was 6 (3-9) days, and the median of onset-diagnosis interval of the secondary cases was 5 (3-8) days. Conclusions Epidemic clusters of COVID-19 were common in many cities outside Wuhan and Hubei. Close contact in family was one of the main causes for the spread of household transmission of the virus. After 4 February, the epidemic clusters were mainly caused by the first generation or second generation cases in local areas, and the time for diagnosis became shorter.