Objective To construct Raji-Luc lymphoma cells with CD19 knockout using CRISPR/Cas9 technology and preliminarily validate their immune escape ability.Methods PB-CRISPR-CD19 small guide RNA(sgRNA)plasmids was constructed,the optimal sgRNA sequence was screened,and Raji-Luc cells with pCAG-PBase,PB-CD19 sgRNA,and PB-CRISPR-Cas9 were co-transfected.Stable knockout monoclonal cell lines were screened by flow sorting and limit dilution method and the knockout effect was verified through gene sequence testing.The expression of luciferase on the surface of the cell line was detected by microplate reader,CD19 CAR-T and CD38 CAR-T previously constructed in the laboratory were used as effector cells,and the immune escape ability of Raji-Luc CD19 KO cell line was verified by universal luciferase chemiluminescence method.Results The transfection efficiency of Raji-Luc CD19 KO cells prepared by electro transfection was high,and the knockout efficiency of the two monoclonal cells was more than 99%.There was no significant difference in luciferase expression compared to the original Raji-Luc cells,and CD19 CAR-T cells could not be activated to the kill them.Conclusion Successfully constructed Raji-Luc CD19 KO lymphoma cell line.

Background ¹³⁷Cs in atmospheric aerosol is the product of past nuclear weapon tests and nuclear accidents. When ¹³⁷Cs is released into the atmosphere, it will deposit in dry land and marine environment, causing pollution of soil surface, water, agricultural products, and animal byproducts, and affecting public health. Objective To identify the variation pattern of ¹³⁷Cs activity concentration in aerosol and its correlation with dust concentration in Beijing area from 2017 to 2020. Methods A total of 958 aerosol samples were collected from November 1, 2017 to June 30, 2020 in Beijing with a high volume air sampler at a sampling flow rate about 600 m³·h⁻¹ and a collection time for each sample about 24 h. The activity concentration of ¹³⁷Cs in the aerosol samples was determined with a low-background high-purity germanium γ spectrometer. The dust concentration was calculated using the difference in the mass of the aerosol filter before and after sampling. The detection rate of ¹³⁷Cs and dust concentration in different seasons were compared. Spearman rank correlation test was used to analyze the correlation between ¹³⁷Cs activity concentration and dust concentration. Results From 2017 to 2020, the ¹³⁷Cs activity concentrations of 33 from 958 aerosol samples in Beijing were above the minimum detectable activityconcentration, the overall detection rate of ¹³⁷Cs was 3.4%, and the activity concentration ranged from 1.86 to 45.53 μBq·m⁻³, with a median value of 4.85 μBq·m⁻³. The detection rate of ¹³⁷Cs was highest in spring, followed by autumn, and lowest in winter and summer (8.4%, 3.0%, 1.1%, and 0.5%, respectively). The dust concentration ranged from 0.03 to 1.55 mg·m⁻³, with an average value of 0.18 mg·m⁻³. There was a statistically significant difference in the dust concentrations in spring, summer, autumn, and winter (F=45.51, P<0.05), and the highest value was 0.24 mg·m⁻³ in spring (P<0.05). The ¹³⁷Cs activity concentration was positively correlated with the dust concentration (P<0.05). Conclusion The ¹³⁷Cs activity concentration in aerosol in Beijing from 2017 to 2020 fluctuates within the range of background level, and its activity concentration is highest in spring, followed autumn, and lowest in summer and winter.

Hip surgery is often associated with moderate to severe postoperative pain due to exten-sive invasion.A safe and effective postoperative pain treatment can promote patients'recovering activities and facilitate the early functional recovery of the hip after the surgery.Although the routinely used intrave-nous self-controlled analgesia has an exquisite analgesic effect,it carries the risks of postoperative nausea and emesis,excessive sedation,intestinal paralysis,and even respiratory depression.As an alternative to transversus abdominis plane block(TAPB),lumbar quadratus block(QLB)has been gradually applied to the hip joint surgery.In this paper,we review the anatomical basis and characteristics of QLB and its clini-cal application in hip surgery,in order to provide the postoperative analgesia application of QLB.

Purpose: Radiation-induced dermatitis (RD) is a common side-effect of therapeutic ionizing radiation that can severely affect patient quality of life. This study aimed to develop a risk prediction model for the occurrence of RD in patients with cervical carcinoma undergoing chemoradiotherapy using electronic medical records (EMRs). Methods: Using EMRs, the clinical data of patients who underwent simultaneous radiotherapy and chemotherapy at a tertiary cancer hospital between 2017 and 2022 were retrospectively collected, and the patients were divided into two groups: a training group and a validation group. A predictive model was constructed to predict the development of RD in patients who underwent concurrent radiotherapy and chemotherapy for cervical cancer. Finally, the model's efficacy was validated using a receiver operating characteristic curve. Results: The incidence of radiation dermatitis was 89.5% (560/626) in the entire cohort, 88.6% (388/438) in the training group, and 91.5% (172/188) in the experimental group. The nomogram was established based on the following factors: age, the days between the beginning and conclusion of radiotherapy, the serum albumin after chemoradiotherapy, the use of single or multiple drugs for concurrent chemotherapy, and the total dose of afterloading radiotherapy. Internal and external verification indicated that the model had good discriminatory ability. Overall, the model achieved an area under the receiver operating characteristic curve of .66. Conclusions The risk of RD in patients with cervical carcinoma undergoing chemoradiotherapy is high. A risk prediction model can be developed for RD in cervical carcinoma patients undergoing chemoradiotherapy, based on over 5 years of EMR data from a tertiary cancer hospital.

Objective:To explore the application value of conventional ultrasound(US) in detecting occult bleeding lesions in joints of patients with hemophilia.Methods:Twenty-seven children with severe hemophilia A who received factor Ⅷ prophylactic treatment from March 2017 to April 2018 in Nanfang Hospital, Southern Medical University were enrolled. Clinical data were collected from routine practice accompanied with US examination, hemophilia joint health score (HJHS) 2.1 assessment and the history of hemarthrosis of both ankles, elbows and knees at baseline and at 1 year follow-up.Results:A total of 157 joints of 27 patients were followed up for one year. During the treatments, 74 joints were found without history of hemarthrosis, among which the total US score of 14(14/74, 18.9%) joints increased, and HJHS 2.1 score of 11(11/74, 14.9%) joints developed. There was no significant change in the HJHS 2.1 score of 108 joints, among which 25 joints (25/108, 23.1%) had an increase in the total US score.However, there was no significant change in the total US score for 93 joints, of which 9(9/93, 9.7%) had an increase in HJHS 2.1 score.Conclusions:US examination can sensitively detect hemophilia arthropathy in joints without history of hemarthrosis and without obvious progression of arthropathy by HJHS 2.1 examination, and provide clinicians with more accurate joint information.

Objective:To construct the prediction model of death risk of Stanford type A aortic dissection (AAD) based on Cox proportional risk regression model.Methods:AAD patients who were diagnosed and received surgical treatment admitted to the department of cardiothoracic surgery of Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology from January 1st, 2019 to April 30th, 2020 were enrolled. The general situation, clinical manifestations, pre-hospital data, laboratory examination and imaging examination results of the patients were collected. The observation period was up to the death of the patients or ended on April 30th, 2021. They were divided into the model group and the verification group according to the ratio of 7∶3. Lasso method was used to screen prognostic variables from the data of the modeling group, and multivariate Cox regression analysis was included to construct the AAD death risk prediction model, which was displayed by nomogram. The receiver operator characteristic curve (ROC curve) was used to evaluate the discrimination of the model, the calibration curve to evaluate the accuracy of the model, and the clinical decision curve (DCA) to evaluate the effectiveness of the model.Results:A totel of 454 patients with AAD were finally included, and the mortality was 19.4% (88/454). Lasso regression analysis was used to screen out 10 variables from the data of 317 patients in the model group, and the prediction model of death risk was constructed: 0.511×abdominal pain+1.061×syncope+0.428×lower limb pain/numbness-0.365×emergency admission-1.933×direct admission-1.493×diagnosis before referral+0.662×preoperative systolic blood pressure (SBP) < 100 mmHg (1 mmHg = 0.133 kPa)+0.632×hypersensitivity cardiac troponin I (hs-cTnI) > 34.2 ng/L+1.402×De Bakey type+0.641× pulmonary infection+1.472×postoperative delirium. The area under the ROC curve (AUC) and 95% confidence interval (95% CI) of the AAD death risk prediction model were 0.873 (0.817-0.928), and that of the verification group was 0.828 (0.740-0.916). DCA showed that the net benefit value of the model was higher. The calibration curve showed that there was a good correlation between the actual observation results and the model prediction results. Conclusion:The AAD death risk prediction model based on abdominal pain, syncope, lower limb pain/numbness, mode of admission, diagnosis before referral, preoperative SBP < 100 mmHg, hs-cTnI > 34.2 ng/L, De Bakey type , pulmonary infection, and postoperative delirium can effectively help clinicians identify patients at high risk for AAD, evaluate their postoperative survival and timely adjust treatment strategies.

Objective: To study the output dose of external beam radiotherapy in Beijing and analyze the cause of output dose  deviation.  Guidance  is  given  on  the  selection  of  measurement  methods,  calculation  formulas  and  parameters. Methods: During 2020, the output doses of medical electron accelerators, tomotherapy, cyberknife, and γ-knife were meas- ured based on the water absorbed dose calibration factor combined with the requirements in the IAEA TRS398 and TRS483 reports, and low-energy X-rays were measured based on the air kerma calibration factor combined with the requirements in the AAPM protocal. The output dose was measured and compared with the preset irradiation value. Results　: A total of 59 external beam radiation therapy equipments’s output dose were investigated.2 of the 49 accelerators’s absolute value of relative deviation between the system indication value and the measurement exceeded 3%; the deviation of other external beam radiotherapy equipments was less than 3%. Conclusion　 A unified method and multi-party quality control can ensure the accuracy of the output dose. In the routine operation of the equipment, the hospital should strictly implement periodic inspections to ensure accurate and reliable radiotherapy.

Objective To investigate the intervention effect of Xuebijing Injection on the differentiation of bone marrow hematopoietic stem cells (HSC) in septic mice. Methods Fifty-four male C57BL/6N mice were randomly divided into three groups: normal control group, model group and Xuebijing group, each group with 18 mice. The mouse models of sepsis were duplicated by intra-peritoneal injection of 10 mg/kg E. coli lipopolysaccharide (LPS) method. Starting from the day of modeling, Xuebijing Injection 20 mL/kg was intravenously injected into the tail vein in Xuebijing group, once a day for consecutive 4 days; the normal control and model groups were intravenously injected with normal saline at the same dose and site for 4 days. The bone marrow cells of the femur and tibia of the mice were isolated after 4 days of various treatments in the three groups, and the proportions of bone marrow HSC Lin-Sca-1+c-Kit+ (LSK) and hematopoietic progenitor cells Lin-Sca-1-c-Kit+ (LS-K) of each group were detected by flow cytometry. Results Finally, 14 mice were included in the normal control group, 17 in the model group, and 12 in the Xuebijing group. With the prolongation of time, the body weight of the normal control group gradually increased, the body masses of the model group and the Xuebijing group were decreased first and then increased, reaching a peak at 96 hours after the model was established, but they were still significantly lower than the body mass of normal control group (g: 19.81±0.27, 19.58±0.39 vs. 22.23±0.30, both P < 0.05). Compared with the normal control group, the proportions of LSK, LS-K, long-term HSC (LT-HSC), and megakaryocyte-erythroid progenitor cells (MEP) were all significantly increased in the model group [LSK: (16.62±1.28)% vs. (12.89±0.83)%, LS-K: (44.77±1.77)% vs. (30.34±0.90)%, LT-HSC: (6.88±0.48)% vs. (1.83±0.24)%, MEP: (13.89±1.26)% vs. (9.38±0.66)%, all P < 0.05], the proportion of multipotential progenitor cells (MPP) was significantly decreased [(2.41±0.34)% vs. (5.99±0.59)%, P < 0.05]. Compared with the model group, the LSK and myeloid progenitor (CMP) of the Xuebijing group were significantly reduced [LSK: (12.25±0.69)% vs. (16.62±1.28)%, CMP :(0.31±0.05)% vs. (0.55±0.13)%, both P < 0.05], and LS-K, LT-HSC, MEP showed a decreasing trend [LS-K: (42.75±2.48)% vs. (44.77±1.77)%, LT-HSC:(5.98±0.70)% vs. (6.88±0.48)%, MEP: (10.94±1.36)% vs. (13.89±1.26) %], but the differences were not statistically significant (all P > 0.05). There were no significant differences in the proportions of short-term HSC (ST-HSC) and granulocyte-macrophage progenitor cells (GMP) among the three septic groups (all P > 0.05). Conclusion Xuebijing Injection can improve the differentiation function of bone marrow cells in septic mice, which may be possibly related to the inhibition of pathological proliferation of bone marrow hematopoietic stem cells and progenitor cells in septic mice by Xuebijing Injection.

Objective To evaluate the effect of clemastine fumarate on Toll-like receptor 4/phosphatidylinositol-3-kinase/serine-threonine kinase (TLR4/PI3K/Akt) signaling pathway during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9C2 cells of rats cultured in vitro were seeded in culture wells or dishes at a density of 1×105 cells/ml and divided into 3 groups (n=11 each) by using a random number table method:control group (group C),H/R group and clemastine fumarate group (CF group).Cardiomyocytes were exposed to 5％ CO2-95％ N2in a low-glucose DMEM medium at 37℃ for 4 h followed by 4 h reoxygenation.At 4 h of reoxygenation,the cell viability was detected by CCK-8 assay,the ultrastructure was observed with a transmission electron microscope,the expression of TLR4,PI3K,phosphorylated Akt (p-Akt) and caspase-3 was detected by Western blot,and the expression of TLR4,PI3K and caspase-3 was detected by immunofluorescence.Results Compared with group C,the cell viability was significantly decreased,the expression of TLR4 and caspase-3 was up-regulated,and the expression of PI3K and p-Akt was down-regulated in group H/R (P＜0.05).Compared with group H/R,the cell viability was significantly increased,the expression of TLR4 and caspase-3 was down-regulated,the expression of PI3K and p-Akt was up-regulated (P＜0.05),and the mitochondrial damage was significantly attenuated in group CF.Conclusion The mechanism by which clemastine fumarate alleviates H/R injury to rat cardiomyocytes may be related to inhibiting TLR4 expression and activating PI3K/Akt signaling pathway.

Objective To investigate the influences of herpes simplex virus 1 and 2 ( HSV1 and HSV2) infection on the expression of signaling molecules associated with innate immune response in respira-tory and vaginal epithelial cells for bettering understanding of HSV infection and pathological characteristics in the primary infection site, namely mucosal epithelial tissues. Methods KMB17 and VK2 cells were in-fected with HSV. Changes in cell morphology and inner structure after HSV infection were observed under optical microscope and scanning electron microscope, respectively. Viral proliferation in KMB17 and VK2 cells was detected by plaque assay, microcytopathic assay and real-time quantitative PCR. Expression of sig-naling molecules associated with innate immune response in virus-infected KMB17 and VK2 cells were ana-lyzed by real-time quantitative PCR. Results Both HSV1 and HSV2 could infect KMB17 and VK2 cells, and cause damage to cell morphology and inner structure after 12 hours. Both of the two viruses formed simi-lar plaque on the single layer of KMB17 and VK2 cells, although HSV2 proliferated slower than HSV1. There were differences in the expression of signaling molecules associated with innate immune response in-duced by the two viruses in KMB17 and VK2 cells. Conclusion Both HSV1 and HSV2 could infect and proliferate in epithelial cells ( KMB17 and VK2 cells) . Although there were slight differences in viral prolif-eration between them, significant differences in the expression of signaling molecules associated with innate immune response induced by the two viruses were observed.