With the rapid advancement of synthetic biology, CRISPR-Cas systems have emerged as a powerful tool for gene editing, demonstrating significant potential in various fields, including medicine, agriculture, and industrial biotechnology. This review comprehensively summarizes the significant progress in applying artificial intelligence (AI) technologies to the design, mining, and modification of CRISPR-Cas systems. AI technologies, especially machine learning, have revolutionized sgRNA design by analyzing high-throughput sequencing data, thereby improving the editing efficiency and predicting off-target effects with high accuracy. Furthermore, this paper explores the role of AI in sgRNA design and evaluation, highlighting its contributions to the annotation and mining of CRISPR arrays and Cas proteins, as well as its potential for modifying key proteins involved in gene editing. These advancements have not only improved the efficiency and precision of gene editing but also expanded the horizons of genome engineering, paving the way for intelligent and precise genome editing.
CRISPR-Cas Systems/genetics* ; Artificial Intelligence ; Gene Editing/methods* ; RNA, Guide, CRISPR-Cas Systems/genetics* ; Machine Learning ; Humans ; Genetic Engineering/methods* ; Synthetic Biology

Objective To explore the efficacy of precise surface positioning method in percutaneous screw fixation for Jones fracture.Methods Data of 46 patients with Jones fractures admitted to our hospital from September 2021 to December 2023 were retrospectively analyzed.According to the different treatment methods,the patients were divided into screw groups(24 patients fixed by percutaneous screws)and tension band groups(22 patients fixed by incision and reset tension band).The general conditions,follow-up time,surgical time,hospitalization time,fracture reduction quality,postoperative complete weight-bearing time,fracture healing time,and complications in the two groups were recorded and compared.The American Association of Foot and Ankle Surgery(AOFAS)system was used to assess the recovery of foot function.Results There was no statistically significant difference in the general data before the operation between the two groups of patients(P＞0.05).The follow-up time of the two groups was 6 to 24 months.Compared with the tension band group,the screw group had shorter operation time[(32.96±5.35)min vs.(47.27±9.42)min],shorter hospitalization time[(5.75±1.18)d vs.(7.68±2.05)d],earlier postoperative complete weight-bearing time[(6.47±0.73)w vs.(7.29±1.66)w],faster fracture healing time[(3.31±0.68)months vs.(4.29±1.29)months],higher AOFAS score[(89.13±2.62)vs.(79.59±4.49)](all P＜0.05).The tension band group of fracture reduction quality was better than the screw group(P＜0.05).The quality of fracture reduction during the operation in the tension band group was better than that in the screw group(P＜0.05).In terms of complications:no complications occurred in the screw group,one patient in the tension band group developed infection,and one patient had a loose Kirchner needle.Conclusion Precise surface positioning method in percutaneous screw fixation for Jones fracture is a safe and effective method with precise positioning,simple operation,small trauma,and rapid rehabilitation,which is worthy of clinical promotion,but the indications need to be strictly controlled according to the type of fracture.

Objective To explore the efficacy of precise surface positioning method in percutaneous screw fixation for Jones fracture.Methods Data of 46 patients with Jones fractures admitted to our hospital from September 2021 to December 2023 were retrospectively analyzed.According to the different treatment methods,the patients were divided into screw groups(24 patients fixed by percutaneous screws)and tension band groups(22 patients fixed by incision and reset tension band).The general conditions,follow-up time,surgical time,hospitalization time,fracture reduction quality,postoperative complete weight-bearing time,fracture healing time,and complications in the two groups were recorded and compared.The American Association of Foot and Ankle Surgery(AOFAS)system was used to assess the recovery of foot function.Results There was no statistically significant difference in the general data before the operation between the two groups of patients(P＞0.05).The follow-up time of the two groups was 6 to 24 months.Compared with the tension band group,the screw group had shorter operation time[(32.96±5.35)min vs.(47.27±9.42)min],shorter hospitalization time[(5.75±1.18)d vs.(7.68±2.05)d],earlier postoperative complete weight-bearing time[(6.47±0.73)w vs.(7.29±1.66)w],faster fracture healing time[(3.31±0.68)months vs.(4.29±1.29)months],higher AOFAS score[(89.13±2.62)vs.(79.59±4.49)](all P＜0.05).The tension band group of fracture reduction quality was better than the screw group(P＜0.05).The quality of fracture reduction during the operation in the tension band group was better than that in the screw group(P＜0.05).In terms of complications:no complications occurred in the screw group,one patient in the tension band group developed infection,and one patient had a loose Kirchner needle.Conclusion Precise surface positioning method in percutaneous screw fixation for Jones fracture is a safe and effective method with precise positioning,simple operation,small trauma,and rapid rehabilitation,which is worthy of clinical promotion,but the indications need to be strictly controlled according to the type of fracture.

Objective To analyze the mortality characteristics and impact on life expectancy of the major malignant tumors among Juxian residents in 2023.Methods Collect the full cause of death registration data and population data for resi-dents of Juxian in 2023.The mortality,cause eliminated life expectancy,PYLL and other indicators of major malignant tumors were calculated.Results In 2023,the death rate of malignant tumors among registered residence residents in Juxian County ranked second in the total causes of death,with a crude mortality rate of 159.80/100 000 and a standardized mortality rate of 133.75/100 000.The mortality rate of malignant tumors in men was higher than that in women.The mortality rate of malignant tumors increases with age(P＜0.001).The top 5 deaths from malignant tumors are lung cancer,gastric cancer,liver cancer,colorectal cancer,and esophageal cancer.The life expectancy of residents in Juxian County in 2023 is 80.61 years,with an in-crease of 3.71 years after malignant tumor removal;The top 5 malignant tumors with increased life expectancy after removing ma-jor malignant tumors are lung cancer,gastric cancer,liver cancer,colorectal cancer,and esophageal cancer.The potential life loss rate caused by malignant tumors is 22.38 ‰,with an average potential life loss of 15.38 years.Among the main malignant tumors,lung cancer,liver cancer,stomach cancer,colorectal cancer,and leukemia have a higher potential life loss rate,with leukemia having the highest average potential life loss.Conclusion Lung cancer,digestive system cancer,leukemia and breast cancer in women have a greater impact on life expectancy.Targeted measures should be taken to prevent and control lung cancer,liver cancer and other cancers with high life loss rates.

Objective To analyze the mortality characteristics and impact on life expectancy of the major malignant tumors among Juxian residents in 2023.Methods Collect the full cause of death registration data and population data for resi-dents of Juxian in 2023.The mortality,cause eliminated life expectancy,PYLL and other indicators of major malignant tumors were calculated.Results In 2023,the death rate of malignant tumors among registered residence residents in Juxian County ranked second in the total causes of death,with a crude mortality rate of 159.80/100 000 and a standardized mortality rate of 133.75/100 000.The mortality rate of malignant tumors in men was higher than that in women.The mortality rate of malignant tumors increases with age(P＜0.001).The top 5 deaths from malignant tumors are lung cancer,gastric cancer,liver cancer,colorectal cancer,and esophageal cancer.The life expectancy of residents in Juxian County in 2023 is 80.61 years,with an in-crease of 3.71 years after malignant tumor removal;The top 5 malignant tumors with increased life expectancy after removing ma-jor malignant tumors are lung cancer,gastric cancer,liver cancer,colorectal cancer,and esophageal cancer.The potential life loss rate caused by malignant tumors is 22.38 ‰,with an average potential life loss of 15.38 years.Among the main malignant tumors,lung cancer,liver cancer,stomach cancer,colorectal cancer,and leukemia have a higher potential life loss rate,with leukemia having the highest average potential life loss.Conclusion Lung cancer,digestive system cancer,leukemia and breast cancer in women have a greater impact on life expectancy.Targeted measures should be taken to prevent and control lung cancer,liver cancer and other cancers with high life loss rates.

Objective:To explore the evaluation value of serum levels of positive pentameric protein 3 (PTX3) and creatine kinase isoenzyme MB (CK-MB) on volume load in patients with chronic decompensated heart failure (CDHF).Methods:A total of 300 CDHF patients who visited the Xingtai Central Hospital from July 2019 to July 2022 were selected and divided into a capacity overload group ( n=182) and a non capacity overload group ( n=118) based on their capacity balance level. Two clinical data sets were compared and analyzed. The receiver operating characteristic (ROC) curve was used to analyze the evaluation value of serum PTX3 and CK-MB levels on the volume load of CDHF patients. The clinical disease characteristics of the two groups of patients were analyzed using univariate analysis, and the influencing factors of volume load of CDHF patients were analyzed using logistic regression. A column chart model was constructed and validated. Results:The body mass index (BMI), waist circumference, systolic blood pressure, diastolic blood pressure, fasting blood glucose (FBG), glycosylated hemoglobin (HbA 1c), C-reactive protein (CRP), uric acid (UA), homeostasis model assessment of insulin resistance (HOMA-IR) of patients in the capacity overload group were higher than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The PTX3, CK-MB, pulmonary capillary wedge pressure (PCWP), and CVP levels of patients in the capacity overload group were higher than those in the non-capacity overload group, while albumin, hemoglobin, and hematocrit were lower than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The ROC curve showed that the area under the curve (AUC) of PTX3 and CK-MB for predicting capacity overload in CDHF patients are 0.795 and 0.718, with sensitivity of 86.2% and 83.7%, specificity of 65.4% and 68.6%, respectively, indicating high predictive accuracy; The AUC of the two joint predictions is 0.817, the sensitivity was 92.5%, and the specificity was 70.6%. The prediction accuracy was higher than PTX3 ( Z=3.812, P<0.05) and CK-MB ( Z=3.365, P<0.05). PTX3, CK-MB, albumin, hemoglobin, hematocrit, PCWP, and central venous pressure (CVP) were all influencing factors of volume load status in CDHF patients (all P<0.05). The column chart risk prediction model established based on these factors had high accuracy and strong applicability in clinical treatment. Conclusions:Serum PTX3 and CK-MB levels are influencing factors for volume overload in CDHF patients. A column chart model constructed in combination with indicators such as albumin, hemoglobin, hematocrit, PCWP, and CVP has high predictive value for the volume overload status of CDHF.

Objective To investigate the clinical features and prognosis of hepatocellular carcinoma(HCC).Methods Medical records were collected from 850 HCC patients who were admitted to Hubei Provincial Hospital of Traditional Chinese Medicine from December 2014 to May 2022,and their clinical and prognostic features were analyzed.The chi-square test were used for comparison of categorical data between groups;the Kaplan-Meier method was used to calculate survival time and survival rate,and the log-rank test was used for comparison of survival time based on baseline features.Results Among the 850 HCC patients,male patients accounted for 82.6%,and the median age at initial diagnosis was 58.0(49.0,66.0)years,with the highest proportion of patients aged 50-69 years(59.8%).The patients with HBV infection accounted for the highest proportion of 77.4%;at initial diagnosis,49.2%of the patients had portal vein tumor thrombus,and 20.2%of the patients had extrahepatic metastasis,among which pulmonary metastasis accounted for the highest proportion of 44.2%(76/172).The patients with Barcelona Clinic Liver Cancer(BCLC)stage A(0),B,C,and D HCC accounted for 20.4%,22.5%,41.5%,and 15.6%,respectively.There was a significant difference in the distribution of BCLC stages between different groups based on sex(χ2=16.631,P=0.001),age(χ2=24.261,P=0.019),place of residence(χ2=39.776,P＜0.001),presence or absence of viral hepatitis(χ2=8.338,P=0.040),and presence or absence of regular antiviral therapy before initial diagnosis(χ2=26.140,P＜0.001).Follow-up was performed for 489 patients till death,with a median survival time of 19.99 months(95%confidence interval[CI]:14.86-25.12),and the 1-,3-,5-,and 10-year cumulative survival rates were 60.7%,39.9%,29.4%,and 22.7%,respectively.There was a significant difference in survival time between different groups based on age(χ2=13.452,P=0.009),history of viral hepatitis(χ2=6.123,P=0.013),regular antiviral therapy before initial diagnosis(χ2=15.505,P＜0.001),comorbidity with type 2 diabetes(χ2=9.820,P=0.002),the number of tumors(χ2=57.713,P＜0.001),maximum tumor diameter(χ2=41.862,P＜0.001),portal vein tumor thrombus(χ2=293.909,P＜0.001),extrahepatic metastasis at initial diagnosis(χ2=118.329,P＜0.001),BCLC stage(χ2=465.638,P＜0.001),surgical resection(χ2=78.86,P＜0.001),local treatment(χ2=36.216,P＜0.001),immune checkpoint inhibitor treatment and/or anti-tumor angiogenesis therapy(χ2=7.182,P=0.007),traditional Chinese medicine decoction treatment(χ2=30.050,P＜0.001),and comprehensive treatment regimens(χ2=13.221,P=0.004).Progression-free survival(PFS)was recorded for 259 patients(30.5%),with a median PFS of 10.98 months(95%CI:8.54-13.42).Conclusion HCC patients exhibit epidemiological characteristics in terms of sex,age,place of residence,presence or absence of viral hepatitis,regular antiviral therapy before initial diagnosis,tumor characteristics,treatment modality,and prognosis,with a low early detection rate and a short overall survival time,and therefore,it is urgent to perform early screening,early diagnosis,and early treatment.

Objective:To explore the incidence of secondary hemophagocytic lymphohistiocytosis (sHLH) in elderly patients with severe SARS-CoV-2 infection, and to analyze and summarize its clinical features and risk factors for early identification of high-risk groups.Methods:A retrospective cohort study was conducted. From January to May 2020, No. 960 Hospital of People's Liberation Army, the Second Hospital Affiliated to Cheeloo College of Medicine of Shandong Province, the First Rehabilitation Hospital of Shandong Province, the Public Health Clinical Center Affiliated to Shandong University, and Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine received 248 patients over 60 years old who were diagnosed with severe SARS-CoV-2 infection during their assistance to Hubei or support for diagnosis and treatment of SARS-CoV-2 infection in Shandong Province. The clinical data of patients were collected. According to the hemophagocytic lymphohistiocytosis diagnosis scoring (HScore) criteria, the patients were divided into sHLH group (HScore > 169) and non-sHLH group (HScore < 98). The demographic data, clinical features, laboratory results, the proportion of organ failure and 60-day mortality of patients were collected and compared between the two groups. The risk factors of sHLH and 60-day death were evaluated through binary multivariate Logistic regression analysis in elderly patients with severe SARS-CoV-2 infection. The receiver operator characteristic curve (ROC curve) was plotted to analyze the diagnostic value of indicators only or combined for sHLH.Results:Among 248 elderly patients with severe SARS-CoV-2 infection, 82 patients with incomplete data and untraceable clinical outcomes, and 35 patients with HScore of 98-169 were excluded. Finally, 131 patients were enrolled in the final follow-up and statistics, including 25 patients in the sHLH group and 106 patients in the non-sHLH group. Compared with the non-sHLH group, plasma albumin (ALB), hemoglobin (Hb), lymphocyte count (LYM), platelet count (PLT), fibrinogen (Fib) and prealbumin (PAB) in the sHLH group were significantly reduced, while alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), MB isoenzyme of creatine kinase (CK-MB), serum creatinine (SCr), C-reactive protein (CRP), D-dimer, ferritin (Fer), lactate dehydrogenase (LDH), procalcitonin (PCT), cardiac troponin I (cTnI), triglycerides (TG), interleukin-6 (IL-6), total bilirubin (TBil) were significantly higher. The fever and fatigue in the sHLH group were more severe than those in the non-sHLH group, and the patients in the sHLH group had higher rates of shock, acute kidney injury, liver dysfunction, and cardiac injury than the non-sHLH group. The 60-day mortality of patient in the sHLH group was significantly higher than that in the non-sHLH group [84.0% (21/25) vs. 40.6% (43/106), P < 0.01]. Binary multivariate Logistic regression analysis showed that high Fer [odds ratio ( OR) = 0.997, 95% confidence interval (95% CI) was 0.996-0.998], D-dimer ( OR = 0.960, 95% CI was 0.944-0.977), LDH ( OR = 0.998, 95% CI was 0.997-0.999) and TG ( OR = 0.706, 95% CI was 0.579-0.860) were independent risk factors for sHLH in elderly patients with severe SARS-CoV-2 infection (all P < 0.01), while elevated Fer ( OR = 1.001, 95% CI was 1.001-1.002), LDH ( OR = 1.004, 95% CI was 1.002-1.005) and D-dimer ( OR = 1.036, 95% CI was 1.018-1.055) were independent risk factors for 60-day death of patients (all P < 0.01). The death risk of the sHLH patients was 7.692 times higher than that of the non-sHLH patients ( OR = 7.692, 95% CI was 2.466-23.987, P = 0.000). ROC curve analysis showed that a three-composite-index composed of LDH, D-dimer and TG had good diagnostic value for sHLH in elderly patients with severe SARS-CoV-2 infection [area under the ROC curve (AUC) = 0.920, 95% CI was 0.866-0.973, P = 0.000]. Conclusions:Elderly patients with severe SARS-CoV-2 infection complicated by sHLH tend to be critically ill and have refractory status and worse prognosis. High Fer, LDH, D-dimer and TG are independent risk factors for sHLH, and are highly suggestive of poor outcome. The comprehensive index composed of LDH, D-dimer and TG has good diagnostic value, and can be used as an early screening tool for sHLH in elderly patients with severe SARS-CoV-2 infection.

Objective:To analyze the risk factors for complications of the retromandibular approach in patients with parotid gland posterior and lower pole tumors.Methods:A retrospective analysis was conducted on the clinical data of 140 patients with parotid posterior lower pole tumors admitted to the Xingtai Third Hospital from October 2019 to October 2021. They were divided into two groups based on whether complications occurred: the occurrence group and the non occurrence group. General data of the two groups of patients were collected, including age, gender, course of disease, previous surgical history, number of tumors, tumor length, resection range, facial nerve dissociation, tumor site resection frequency, and fascia preservation; Single factor and logistic multivariate analysis were conducted to determine the risk factors for complications of the posterior retromandibular approach in patients with parotid gland posterior and lower pole tumors.Results:A total of 140 patients with parotid gland posterior lower pole tumors underwent retromandibular approach treatment, with complications occurring in 38 cases (27.14%), including 7 cases of temporary facial paralysis, 10 cases of facial depression, 11 cases of Frey syndrome, 2 cases of fistula, and 8 cases of sensory abnormalities of the greater auricular nerve. Through logistic multivariate analysis, it was found that the number of tumors ≥ 2 ( OR=2.856), the resection range (total resection) ( OR=2.477), the number of surgeries ≥3 ( OR=5.637), facial nerve dissociation ( OR=3.526), and lack of fascia preservation ( OR=2.551) were all risk factors for postoperative complications in patients with parotid posterior pole tumors (all P<0.05). Conclusions:In clinical practice, relevant prevention and treatment measures should be formulated for these high-risk factors to reduce the incidence of postoperative complications.

Objective:To detect the expression levels of laboratory of genetics and physiology 2 (LGP2), retinoic acid inducible gene I (RIG-I) and melanoma differentiation associated gene 5 (MDA5) in children with hand, foot and mouth disease (HFMD), and to explore their possible clinical significance in HFMD.Methods:Fifty children with HFMD, who visited Second Affiliated Hospital of Xi′an Jiao Tong University, Xi ′an Children′s Hospital and Xi ′an Central Hospital from May 2020 to May 2021, were selected as the research subjects, and 20 children with physical examination at the same age during the same period were selected as the control group.Children with HFMD were divided into enterovirus 71 (EV-A71) type and coxsackievirus A6 (CV-A6) type according to the results of pathogen detection, and then divided into mild group and severe group according to the severity of the disease.The relative mRNA expression levels of LGP2, RIG-I and MDA5 in each group, and the correlation among the three proteins were compared and analyzed.Results:Among 50 cases of HFMD, 26 cases were EV-A71 type (16 cases were mild and 10 cases were severe) and 24 cases were CV-A6 type (17 cases were mild and 7 cases were severe). There was no significant difference in age and sex between HFMD group and control group ( P>0.05). The relative expression levels of LGP2 mRNA in EV-A71 and CV-A6 HFMD cases were 2.37(1.78, 3.25)% and 1.88 (1.35, 3.13)%, lower than that in control group [2.97(2.61, 3.55)%]. Only the difference between CV-A6 HFMD children and control group was statistically significant ( Z=-2.310, P=0.021). The relative expression levels of RIG-I mRNA in EV-A71 and CV-A6 HFMD cases were 9.95 (7.79, 14.62)% and 9.78(7.04, 15.83)%, lower than that in control group [18.47(13.00, 21.07)%]. The differences were all statistically significant ( P<0.05). The relative expression levels of MDA5 mRNA in EV-A71 and CV-A6 HFMD cases were 4.41(2.82, 5.99)% and 3.98 (2.18, 7.41)%, lower than that in control group [5.10(3.52, 7.71)%], but the differences were not statistically significant.There were no significant differences in the relative expression levels of the three indicators between the mild and severe groups of children with EV-A71 or CV-A6 HFMD.The expression levels of LGP2, RIG-I and MDA5 mRNA were highly correlated( P<0.001). Conclusion:The relative expression levels of LGP2, RIG-I and MDA5 mRNA in children with HFMD are decreased in different degrees than those in normal children.And there is a correlation among them.