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.

Perinatal depression is a psychological disorder that occurs during pregnancy and within one year of delivery, which can seriously affect the physical and mental health of pregnant and postpartum women, as well as the cognitive and behavioral abilities of offspring, with potential multigenerational effects. Therefore, it is important to identify its potential modifiable risk factors. Phthalic acid esters (PAEs), as common environmental endocrine disruptors, can affect maternal estrogen through multiple mechanisms and are important potential modifiable risk factors for developing maternal perinatal depression. At present, studies on the correlation between PAEs and perinatal depression are still very limited, and the mechanisms by which PAEs affect perinatal depression have not been clarified. Based on existing epidemiological and toxicological studies at home and abroad, the article briefly introduced the characteristics of multiple pathways, high doses, and long-term exposure to maternal PAEs, focused on reviewing the current status of epidemiological studies, pointed out the possible associations between some specific PAEs exposure and elevated risk of perinatal depression. It also summarized the potential roles of hormone-neurotransmitter pathway, inflammation mediation, gene regulation, and other possible mechanisms in the association between exposure to PAEs and perinatal depression. The article concluded with a look at how future research on the association between exposure to PAEs and perinatal depression can be scientifically validated, with a view to providing more high-quality evidence for the scientific prevention of the onset and progression of maternal depressive symptoms.

[Objective]To elucidate the pathogenesis and treatment of neck-shoulder syndrome based on the viewpoint of"liver depression and Qi stagnation"of the theory of five viscera Qi activity.[Methods]From the viewpoint of"liver depression and Qi stagnation"in the theory of five viscera Qi and the etiology and pathogenesis of neck-shoulder syndrome,the theoretical correspondence between the two was discussed,and based on the viewpoint of"liver depression and Qi stagnation",it established the therapeutic principles and acupuncture prescriptions of neck-shoulder syndrome,and cited a case to support it.[Results]The viewpoint of"liver depression and Qi stagnation"is rooted in the liver governing Qi of the five viscera Qi activity theory,taking liver depression and Qi stagnation as the foundation.The pathogenesis of neck-shoulder syndrome is characterized by the mixture of asthenia and sthenia of the specimen,mainly liver dysfunction and Qi stagnation,on the basis of which it causes the formation or combination of stasis,dampness and other excess evils.To analyze the pathogenesis of neck-shoulder syndrome from"liver depression and Qi stagnation",acupuncture treatment should not only pay attention to liver and gallbladder,take into account Qi stagnation at the same time,but also pay attention to its evolution of stasis,dampness and other pathogenic factors,which are closely related to the generation of neck-shoulder syndrome.To guide the treatment of neck-shoulder syndrome with"liver depression and Qi stagnation"and related theories,should take dredging liver and gallbladder and Qi as the key,it is required to start from the acupuncture and moxibustion to harmonize the meridian-collateral Qi activity,and cooperate with the knife-edge needle,and clinical practice supports the rationality of the theory.The medical case listed was neck-shoulder syndrome,and the syndrome was liver depression and Qi stagnation.The main treatment was to dredge liver Qi to relieve pain,and the use of acupuncture and knife-edge needle had obtained better clinical effect.[Conclusion]The view of"liver depression and Qi stagnation"is consistent with the pathogenesis of neck-shoulder syndrome,which opens up a new way for the treatment of neck-shoulder syndrome.

Objective:To investigate the predictive value of neutrophil-lymphocyte ratio (NLR) for Trousseau’s syndrome (TS) in patients with acute multiple cerebral infarctions (AMCI).Methods:The patients with AMCI in Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine from July 2013 to March 2022 were retrospectively enrolled. The demographic and baseline clinical data of patients with TS and those without TS were compared. Multivariate logistic regression analysis was used to determine the independent influencing factors of TS-AMCI, and receiver operating characteristic (ROC) curve was used to evaluate the predictive value of NLR for TS-AMCI. Results:A total of 59 patients with AMCI were enrolled, including 43 males and 16 females, aged 64.9±14.0 years. There were 16 patients in the TS-AMCI group and 43 in the non-TS-AMCI group. The proportions of patients with diabetes mellitus, hypertension and previous stroke or transient ischemic attack in the TS-AMCI group were significantly lower than those in the non-TS-AMCI group (all P<0.05), while the proportion of patients with ischemic heart disease were significantly higher than that in the non-TS-AMCI group ( P<0.05). The proportion of patients with bilateral infarction in the TS-AMCI group was significantly higher than that in the non-TS-AMCI group ( P<0.001). The D-dimer, NLR, white blood cell count, neutrophil count, monocyte count, percentage of neutrophils, total cholesterol and low-density lipoprotein cholesterol in the TS-AMCI group were significantly higher than those in the non-TS-AMCI group (all P<0.001), while the lymphocyte count, lymphocyte percentage, red blood cell count, hemoglobin and hematocrit were significantly lower than those in the non-TS-AMCI group (all P<0.001). Multivariate logistic regression analysis showed that high NLR was an independent predictor of TS-AMCI (odds ratio [ OR] 2.897, 95% confidence interval [ CI] 1.270-6.527; P=0.011), while high hemoglobin was independently negatively correlated with TS-AMCI ( OR 0.839, 95% CI 0.723-0.975; P=0.022). ROC curve analysis showed that the area under the curve of NLR for predicting TS-AMCI was 0.929 (95% CI 0.831-0.979; P<0.001). When the NLR cutoff value was 4.01, the corresponding Youden index was 0.744. At this time, the sensitivity and specificity were 100% and 74.42% respectively. Conclusion:NLR has high predictive value for TS-AMCI.

Objective:To compare the accuracy of two methods in reducing leg length discrepancy(LLD)during hip hemiarthroplasty.Methods:We retrospectively analyzed 89 patients of hip hemiarthroplasty who suffered from femoral neck fracture.There were 47 patients in the new method group(NM), and 42 patients in the traditional method group(traditional method, TM)which comparing the position of the greater trochanter tip and the center of the femoral head.In the NM group, the distance from the center of femoral head to the lesser trochanter(L)and the diameter of femoral head(D)of the healthy side hip were measured on preoperative anteroposterior pelvic X-ray film, and the ratio(R)of D to L was calculated.During operation, the diameter of the femoral head(d)was measured with a caliper, and the distance should be obtained from the center of the femoral head prosthesis to the lesser trochanter according to the ratio R of the healthy side.The difference of postoperative LLD between the two groups and the incidences of |LLD| in each range were compared.Results:In the NM group, the maximum LLD was 11.10 mm and the minimum LLD was -4.0 mm, with an average of(4.4±3.2)mm, 80.9%(38/47)| LLD | < 6 mm, 93.6%(44/47)| LLD | < 10 mm, 6.4%(3/47)| LLD | ≥ 10 mm.In the TM group, the maximum LLD was 13.2 mm and the minimum LLD was -8.3 mm, with an average of (6.2±5.1)mm, 42.9%(18/42)|LLD|<6 mm, 69.0%(29/42)|LLD| <10 mm, 31.0%(13/42)|LLD|≥10mm.The differences of patients of the mean postoperative LLD and the incidences of |LLD| in each range between two the groups were statistically significant( t=-2.036、 χ2=14.629, P=0.046、0.001). Conclusions:The new method is simple, convenient, more accurate and can obtain a more satisfactory LLD compared with the traditional method which refers to the relative position of the great trochanter tip and the center of the femoral head.

Ferroptosis is a type of cell death accompanied by iron-dependent lipid peroxidation, thus stimulating ferroptosis may be a potential strategy for treating gastric cancer, therapeutic agents against which are urgently required. Jiyuan oridonin A (JDA) is a natural compound isolated from Jiyuan

Histone lysine specific demethylase 1 (LSD1) has been recognized as an important modulator in post-translational process in epigenetics. Dysregulation of LSD1 has been implicated in the development of various cancers. Herein, we report the discovery of the hit compound (IC = 3.93 μmol/L) and further medicinal chemistry efforts, leading to the generation of compound (IC = 49 nmol/L, and = 16 nmol/L), which inhibited LSD1 reversibly and competitively with H3K4me2, and was selective to LSD1 over MAO-A/B. Docking studies were performed to rationalize the potency of compound . Compound also showed strong antiproliferative activity against four leukemia cell lines (OCL-AML3, K562, THP-1 and U937) as well as the lymphoma cell line Raji with the IC values of 1.79, 1.30, 0.45, 1.22 and 1.40 μmol/L, respectively. In THP-1 cell line, significantly inhibited colony formation and caused remarkable morphological changes. Compound induced expression of CD86 and CD11b in THP-1 cells, confirming its cellular activity and ability of inducing differentiation. The findings further indicate that targeting LSD1 is a promising strategy for AML treatment, the triazole-fused pyrimidine derivatives are new scaffolds for the development of LSD1/KDM1A inhibitors.

Objective To investigate the impact of the changes of posttreatment karnofsky performance status (KPSpost) on the overall survival (OS) for patients with stage Ⅳ non-small cell lung cancer (NSCLC) underwent concurrent chemoradiation.Methods A total of 279 patients (male 198 and female 81) with histological confirmed stage Ⅳ NSCLC were enrolled in this study with a median age of 58 years old (range 22 to 80 years old).There were 166 cases of squamous carcinoma,87 cases of adenocarcinoma,and 22 cases of unclassified carcinoma,respectively.All enrolled patients received more than 2 cycles of chemotherapy and more than 36 Gy of concurrent radiotherapy.Kaplan-Meier method and Log-rank test were applied to evaluate OS.Multivariate analyses were carried out by the Cox proportionalhazard model.Chi-square test and logistic regression analysis were used to explore the related factors of KPSpost.Results There were 198 patients with improved KPSpost and 81 patients with decreased KPSpost,respectively.Univariate and multivariate analyses indicated that the improvement of KPSpost was associated with longer OS.Logistic regression analysis showed that the improvement of KPSpost was positively related with treatment of more than 4-6 cycles chemotherapy concurrent with over 63 Gy radiation to primary tumor.The improvement of KPSpost also correlated positively with disease control rate (DCR),but negatively with PLT toxicity and radiation esophagitis.Conclusions KPSpost was an independent prognostic factor of OS for patients with stage Ⅳ NSCLC underwent concurrent chemoradiation.Chemotherapy of 4-6 cycles and concurrent over 63 Gy radiotherapy dose to primary tumor,as well as DCR were positive factors for KPSpost improvement.However,stage 3-4 PLT toxicities and radiation esophagitis decreased the KPSpost.

Objective To evaluate the feasibility and safety of the robot - assisted system YunSRobot for remote manipulation endoscopy. Methods When the master of YunSRobot was installed in the gastroenterology office in Chinese PLA General Hospital, the robot slave and upper gastrointestinal simulation model (Takahashi Lm103,Japan) were installed at the same time in the State Key Laboratory of Robotics, Shenyang Institute of Automation. Three physicians were trained to operate the master robotics and performed gastroscopy on the simulation model based on network cloud. Each physician performed 3 procedures of oesophagogastroduodenoscopy (EGD) by YunSRobot using traditional manual endoscopy, on-site operating mode, and remote manipulation mode, respectively. The operating time, lumenal anatomic exposure,man-machine interaction and other parameters were recorded. Results The number of standard pictures obtained by traditional manual endoscopy group, on-site operating group and remote manipulation group were 39.9±0.3, 39.8±0.4, 39.9±0.3, respectively. The images of all five lesions could be obtained by each operation. The operating time in the duodenum of remote group was longer than that of on-site group, with average time (78.2±16.0)s vs. (68.9±15.8)s (P=0.021) respectively. As to the operating time on other parts or total time, all three groups were comparable. Although there was a mean delay of (572.1±48.5) ms in remote operation group, the operation was still smooth. However, compared with on-site group, the percentage of clear view time in the duodenum was significantly shortened in remote group: [(77.8±8.2)% vs. (83.9 ± 6.4)% , P=0.024]. Statistically significant difference was detected in percentage of clear view time neither in other sites, nor was in the total operating time between two groups. The operating time in each part of remote group was obviously longer than that of manual group as followings, pharyngeal (27.3±4.2) s vs. (9.2±1.3)s (P<0.001), esophageal (29.7±6.4)s vs. (19.3±1.6)s (P=0.004), stomach (56.7±17.0)s vs. (40.3±7.0)s (P=0.003), pylorus (20.2±5.5)s vs. (9.3±1.3)s (P<0.001), duodenum (78.2±16.0)s vs. (29.3±5.6)s (P<0.001). Thus the total operating time was also longer in remote group as (559.0±87.2)s vs. (253.1±16.6)s (P<0.001). The respective time in pharynx, esophagus, stomach, pylorus, duodenum, or the overall time was all longer in remote group than that in manual group. Conclusions The soft endoscopy robot YunSRobot has satisfactory safety and stability. Remote upper gastrointestinal endoscopy can be completed based on common network and an endoscope simulation model with smooth operation. The inspection time by YunSRobot robot per part and the overall time are longer than those of manual operation on site, still, remote operating time meets the standard of upper gastrointestinal endoscopy.

Objective: To study the effects of estrogen on the degenerative changes of condylar cartilage and subchondral bone in rats. Methods: 18 female SD rats aged 6 weeks were divided into control(C),unilateral anterior cross-bite(UAC) and UAC treated with estrogen(UAC + E) groups(n = 6). UAC metal prosthesis was cemented to the left incisors of maxilla and mandible of the rats in group UAC and UAC + E. Rats in UAC + E group were given pexitoneal injection of 80 μg 17β-estradiol per day. The rats in group C were untreated. Animal were sacrificed at the 4th weeks. The micro-structure of subchondral bone was observed by Micro-CT scanning. HE staining,Safranin O staining,immunohistochemical staining,TUNEL staining and TRAP staining for the observation of pathological changes of histomorphology,extracellular matrix,chondrocyte apoptosis in condylar cartilage,and osteoclasts number in subchondral bone. Results: UAC and UAC + E group showed evident osteoarthritis(OA)-like lesions. Compare with UAC group,there was a significant decrease in the expression of proteoglycan(P ＜ 0. 05),type Ⅱ collagen(P ＜ 0. 01),and a significant increase in the number of apoptotic chondrocytes(P ＜ 0. 01) in UAC + E group. As for subchondral bone,the BV/TV,Tb. Th parameters in C and UAC + E groups were significant higher than in UAC group(P ＜ 0. 01),while the BS /BV,Tb. N,Tb. Sp parameters and the osteoclasts number in C and UAC + E groups were significant fewer than in UAC groups(P ＜ 0. 01). There was no significant difference in bone ultra-parameters and osteoclasts number between C and UAC + E groups(P＞ 0. 05). Conclusion: In the model of rat TMJOA induced by unilateral anterior crossbite prosthesis,supra-physiological level of estrogen can reverse bone loss in subchondral bone,but accelerate the degradation of condylar cartilage.