Objective To investigate the cellular characteristics in the development of retinal tumors induced by MY-CN overexpression,and to find the key proteins that play an important role in this process.Methods T-MYCN Y1 and Y2 cells cultured in vitro for 1 year and 2 years were selected for the experiment.The WERI-RB-1 cell line obtained from Shanghai Cell Bank of Chinese Academy of Sciences was taken as the mature retinoblastoma(RB)control(WERI-Rb-1 group).Untreated normal human retinal tissues were used as the control group.Cells were inoculated into a 24-well plate at a density of 1 × 104 cells per well,and the cell morphology was observed under a microscope and images were collected.Cell count was performed on the 4th,8th and 12th day after inoculation.Immunofluorescence staining was employed to de-tect the expression of the proliferation marker Ki67 and cone cell marker L/M opsin in each group.Quantitative real time polymerase chain reaction(qRT-PCR)was utilized to assess the expression of MYCN,CCNB1,CDK1,SYK,and RXRγ in cells across all groups.Proteomics analysis was performed to detect variations in protein expression profiles across groups and to identify key regulatory genes.Subsequently,cell growth and proliferation were evaluated after OTX2 knockdown by lentiviruses.Results The growth curve showed that there were(59.17±4.01)× 104 cells in the T-MYCN Y1 group,(85.14±4.54)× 104 cells in the T-MYCN Y2 group and(100.73±1.99)× 104 cells in the WERI-Rb-1 group on the 12th day of in-vitro culture(all P＜0.05).Compared with those in the control group,the positive cell rates of Ki67 and L/M opsin increased in T-MYCN Y1,T-MYCN Y2 and WERI-Rb-1 groups(all P＜0.001).The relative expression levels of MYCN mR-NAs in T-MYCN Y1,T-MYCN Y2 and WERI-Rb-1 groups were significantly higher than those in the control group(all P＜0.001).The relative expression levels of CCNB1 and CDK1 mRNAs also increased significantly in the three groups,com-pared with those in the control group(all P＜0.01).There was no significant difference in the relative expression of SYK mRNAs between T-MYCN Y1 and control groups(P＞0.05).The relative expression level of RXRγ mRNAs was significantly higher in the T-MYCN Y1 group than that in the control group(P＜0.05).The relative expression levels of SYK and RXRγmRNAs in T-MYCN Y2 and WERI-Rb-1 groups were significantly increased,compared with those in the control group(all P＜0.05).GO enrichment analysis showed that these differentially-expressed proteins were enriched into protein complex assembly and mitochondria pathways.KEGG enrichment analysis revealed that the enriched pathways of these differential-ly-expressed proteins included carbon metabolism,metabolic pathway and fatty acid degradation.Both proteomics and Western blot analyses indicated that OTX2 expression levels were the highest in the WERI-Rb-1 group,second highest in the T-MYCN Y2 group,and the lowest in the T-MYCN Y1 group(all P＜0.01).According to qRT-PCR,the expression levels of OTX2 mRNAs in T-MYCN Y1,T-MYCN Y2 and WRI-Rb-1 cells with OTX2 knockdown were lower than those in empty vec-tor controls(all P＜0.05).The growth curve of cells showed that after knocking down OTX2 in the three groups of cells,both cell growth and proliferation ability decreased significantly(all P＜0.001).Conclusion During the development of retinal tumors induced by MYCN overexpression,cells gradually exhibit characteristics similar to WERI-Rb-1 cells,which are a typical RB cell line.OTX2 plays an important role in MYCN amplification or the growth and proliferation of highly ex-pressed RB.Targeting OTX2 may become a new research direction for the treatment of RB.

Objective To investigate the cellular characteristics in the development of retinal tumors induced by MY-CN overexpression,and to find the key proteins that play an important role in this process.Methods T-MYCN Y1 and Y2 cells cultured in vitro for 1 year and 2 years were selected for the experiment.The WERI-RB-1 cell line obtained from Shanghai Cell Bank of Chinese Academy of Sciences was taken as the mature retinoblastoma(RB)control(WERI-Rb-1 group).Untreated normal human retinal tissues were used as the control group.Cells were inoculated into a 24-well plate at a density of 1 × 104 cells per well,and the cell morphology was observed under a microscope and images were collected.Cell count was performed on the 4th,8th and 12th day after inoculation.Immunofluorescence staining was employed to de-tect the expression of the proliferation marker Ki67 and cone cell marker L/M opsin in each group.Quantitative real time polymerase chain reaction(qRT-PCR)was utilized to assess the expression of MYCN,CCNB1,CDK1,SYK,and RXRγ in cells across all groups.Proteomics analysis was performed to detect variations in protein expression profiles across groups and to identify key regulatory genes.Subsequently,cell growth and proliferation were evaluated after OTX2 knockdown by lentiviruses.Results The growth curve showed that there were(59.17±4.01)× 104 cells in the T-MYCN Y1 group,(85.14±4.54)× 104 cells in the T-MYCN Y2 group and(100.73±1.99)× 104 cells in the WERI-Rb-1 group on the 12th day of in-vitro culture(all P＜0.05).Compared with those in the control group,the positive cell rates of Ki67 and L/M opsin increased in T-MYCN Y1,T-MYCN Y2 and WERI-Rb-1 groups(all P＜0.001).The relative expression levels of MYCN mR-NAs in T-MYCN Y1,T-MYCN Y2 and WERI-Rb-1 groups were significantly higher than those in the control group(all P＜0.001).The relative expression levels of CCNB1 and CDK1 mRNAs also increased significantly in the three groups,com-pared with those in the control group(all P＜0.01).There was no significant difference in the relative expression of SYK mRNAs between T-MYCN Y1 and control groups(P＞0.05).The relative expression level of RXRγ mRNAs was significantly higher in the T-MYCN Y1 group than that in the control group(P＜0.05).The relative expression levels of SYK and RXRγmRNAs in T-MYCN Y2 and WERI-Rb-1 groups were significantly increased,compared with those in the control group(all P＜0.05).GO enrichment analysis showed that these differentially-expressed proteins were enriched into protein complex assembly and mitochondria pathways.KEGG enrichment analysis revealed that the enriched pathways of these differential-ly-expressed proteins included carbon metabolism,metabolic pathway and fatty acid degradation.Both proteomics and Western blot analyses indicated that OTX2 expression levels were the highest in the WERI-Rb-1 group,second highest in the T-MYCN Y2 group,and the lowest in the T-MYCN Y1 group(all P＜0.01).According to qRT-PCR,the expression levels of OTX2 mRNAs in T-MYCN Y1,T-MYCN Y2 and WRI-Rb-1 cells with OTX2 knockdown were lower than those in empty vec-tor controls(all P＜0.05).The growth curve of cells showed that after knocking down OTX2 in the three groups of cells,both cell growth and proliferation ability decreased significantly(all P＜0.001).Conclusion During the development of retinal tumors induced by MYCN overexpression,cells gradually exhibit characteristics similar to WERI-Rb-1 cells,which are a typical RB cell line.OTX2 plays an important role in MYCN amplification or the growth and proliferation of highly ex-pressed RB.Targeting OTX2 may become a new research direction for the treatment of RB.

Objective:To explore the effect of esketamine on inflammatory cytokines and myocardial injury markers in children undergoing living-donor liver transplantation (LT).Methods:Considering the inclusion criteria, 50 children with biliary atresia were selected for living donor LT. They were equally randomized into two groups of control (C) and esketamine (E) (25 cases each). Esketamine 0.5 mg/kg was administered to group E during induction and continued at a dose of 0.5 mg·kg –1·h -1 after an induction of anesthesia. Group C provided the same dose of 0.9% sodium chloride injection during induction and then continued to pumping until the end of the procedure. Basic profiles of two groups were recorded. Hemodynamic parameters, such as heart rate (HR), mean arterial pressure (MAP) and central venous pressure (CVP), were monitored at 5 min of anesthesia induction (T 0), 30 min of anhepatic phase (T 1), immediately after repercussion (T 2), 30 min of neohepatic phase (T 3) and end of surgery (T 4) in both groups. Central venous blood samples were collected at T 0, T 1, T 3 and T 4. Serum levels of cardiac troponin I (cTnI), creatine kinase isoenzyme-MB (CK-MB) ,tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) were measured. The incidence of adverse cardiac events, postoperative mechanical ventilation time, ICU stay and hospitalization length were compared. Results:As compared with T 0, mean arterial pressure (MAP) at T 2 declined markedly in group E [(48.6±12.7) mmHg (1 mmHg=0.133 kPa) vs (55.6±10.7) mmHg, P<0.001] and C [(39.3±8.0) mmHg vs (53.2±9.4) mmHg, P<0.001 ] ;As compared with T 0, the TNF-α and IL-6 spiked at T 3 in group C [169.0 (207.1) ng/L vs 43.8 (26.4) ng/L, (132.63±51.75) ng/L vs (51.79±17.83) ng/L, P<0.001] and E [78.5 (138.8) ng/L vs 43.8 (26.4) ng/L, (87.44±32.17) ng/L vs (51.79±17.83) ng/L, P<0.001 ] ; In group C, the concentration of myocardial injury markers CK-MB and cTnI rose at T 3/T 4 compared with T 0[T 3 vs T 0: 5.7 (5.4) μg/L vs 4.0 (3.5) μg/L, 0.09 (0.08) μg/L vs 0.02 (0.02) μg/L; T 4 vs T 0: 5.3 (5.0) μg/L vs 4.0 (3.5) μg/L, 0.07 (0.08) μg/L vs 0.02 (0.02) μg/L, P<0.001 ]. In group E, the levels of CK-MB and cTnI were higher at T 3/T 4 than those at T 0[T 3 vs T 0: 7.0 (5.0) μg/L vs 4.6 (2.1) μg/L, 0.06 (0.09) μg/L vs 0.03 (0.04) μg/L; T 4 vs T 0: 5.4 (4.9) μg/L vs 4.6 (2.1) μg/L, 0.03 (0.06) μg/L vs 0.03 (0.04) μg/L; P<0.001]. Compared with group C, the MAP of E rose at T 1/T 2/T 3 [(58.8±10.3) mmHg vs (53.3±8.6) mmHg, P=0.048; (48.6±12.7) mmHg vs (39.3± 8.0) mmHg, P=0.003; (55.8±7.4) mmHg vs (51.5±7.3) mmHg, P=0.044]. Compared with group C, TNF-α and IL-6 decreased in E at T 3/T 4[T 3: 78.5 (138.8) ng/L vs 169.0 (207.1) ng/L, P=0.010; (87.44±32.17) ng/L vs (132.63±51.75) ng/L, P=0.017. T 4: 62.3 (118.3) ng/L vs 141.3 (129.2) ng/L, P=0.001; (74.34±26.38) ng/L vs (100.59±30.40) ng/L, P=0.002]. Compared with group C, cTnI decreased in E at T 3/T 4[0.06 (0.09) μg/L vs 0.09 (0.08) μg/L, P=0.014; 0.03 (0.06) μg/L vs 0.07 (0.08) μg/L, P=0.003]. Compared with group C, the mechanical ventilation time in group E decreased [195 (120) min vs 315 (239) min, P<0.001]. Compared with group C, the incidence of severe hypotension [16%(4/25) vs 48% (12/25), P=0.015 ], bradycardia [12% (3/25) vs 36 % (9/25), P=0.047 ], myocardial ischemia [4 % (1 /25) vs 24 % (6/25), P=0.042 ] and premature ventricular contractions [0 vs 4 %(1/25), P=0.312 ] decreased in group E. Conclusion:Intraoperative dosing of esketamine may suppress inflammatory reactions and alleviate perioperative myocardial injury in children undergoing living-donor LT.

Esketamine is a spin isomer of ketamine,which has the triple effect of sedation,analge-sia and amnesia,and is superior to ablative ketamine in terms of efficacy,controllability.It has been widely used in anesthesia,emergency and critical care in Europe and America,and is mostly used for sedation,analgesia and antidepressant in China.Esketamine is used in cardiac surgery to maintain stable hemodynam-ics,reduce the secretion of inflammatory factors and relieve postoperative pain.Its sympathomimetic effect allows it to be used for the induction of anesthesia in patients with hemodynamic instability and acute heart attack.This paper reviews the recent advance in the clinical value and limitations of esketamine in the perio-perative period of cardiovascular surgery and provides a reference for clinicians to use esketamine in the perioperative period of cardiovascular surgery.

Objective:To evaluate the effect of s-ketamine on perioperative myocardial injury in patients undergoing liver transplantation.Methods:This was a prospective randomized controlled study. Sixty American Society of Anesthesiologists Physical Status classification Ⅲ or Ⅳ patients, aged 18-64 yr, with New York Heart Association classⅠ-Ⅲ, undergoing elective liver transplantation with general anesthesia in our hospital from May to October 2023, were divided into 2 groups ( n=30 each) using a random number table method: s-ketamine group (group S) and control group (group C). In group S, s-ketamine was intravenously injected at a dose of 0.5 mg/kg after induction of anesthesia, followed by an infusion of 0.5 mg·kg -1·h -1 until the end of surgery. The equal volume of normal saline was given instead in group C. Central venous blood samples were collected after induction of anesthesia (T 0), at 30 min of anhepatic phase (T 1), 30 min of neopepatic phase (T 2), abdominal closure (T 3), 24 h after operation (T 4) and 72 h after operation (T 5) for determination of the concentrations of serum high-sensitivity cardiac troponin I, creatine kinase-MB isoenzyme, N-terminal pro-B-type natriuretic peptide, tumor necrosis factor-α, interleukin-6 (IL-6), IL-10 and high-mobility group protein B1 by enzyme-linked immunosorbent assay. The occurrence of adverse cardiac events during surgery and within 24 h after surgery, postoperative mechanical ventilation time, time of intensive care unit stay, and postoperative length of hospital stay were recorded. Results:Compared with group C, the concentrations of serum high-sensitivity cardiac troponin I, creatine kinase-MB isoenzyme, N-terminal pro-B-type natriuretic peptide, tumor necrosis factor-α and IL-6 at T 2-5 and high-mobility group protein B1 at T 2-4 were significantly decreased, the concentrations of serum IL-10 were increased at T 2-5, the incidence of myocardial ischemia was decreased, the mechanical ventilation time was shortened ( P<0.05), and no significant change was found in the time of intensive care unit stay and postoperative length of hospital stay in S group ( P>0.05). Conclusions:Intraoperative usage of s-ketamine can inhibit the inflammatory responses and reduce perioperative myocardial injury in the patients undergoing liver transplantation.

Objective:To classify the ultrasound features of ovarian tumors by modified gynecology imaging reporting and data system(mGI-RADS), and explore the clinical value of mGI-RADS in differentiating of benign and malignant tumors.Methods:The 242 ultrasound images of the adnexal mass from 221 patients with ovarian tumor who underwent ultrasound scan and proceeded surgeries in the Second Affiliated Hospital of Harbin Medical University from September 2017 and December 2019 were involved in the retrospective analysis and compared with the pathological results. According to the latest ultrasound terminology for adnexal masses proposed by the International Ovarian Tumor Analysis (IOTA), the adnexal masses were classified. The suspeted malignant sings were screened for mGI-RADS. Using GI-RADS and mGI-RADS classification systems, all the adnexal masses were double-blindly classified by two senior doctors and their diagnostic efficiencies were evaluated.Results:Among the suspected malignant signs proposed by IOTA, resistance index (RI)<0.5, central blood flow signal, ascites, irregular cystic wall with uneven thickness separation, mass composition (solid component ≥30% and silent shadow attenuation), and papillary structure≥7 mm, showed retatively higher OR values(14.282, 10.372, 9.653, 8.832, 5.851, 4.506, respectively. Using GI-RADS and mGI-RADS classification systems, the diagnostic consistency by the two senior doctors in differentiating benign and malignant ovarian neoplasms was good(Kappa=0.767, P<0.05). Grade Ⅳ was divided into three subtypes (Ⅳa, Ⅳb, Ⅳc) by the mGI-RADS, and their positive predictive values for malignant ovarian masses were 69.0%, 90.4% and 92.9% respectively. When a cut-off value >mGRⅣ was used to distinguish the benign and malignant adnexal tumors, the AUC was the largest, and its sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 93.4%, 93.1%, 90.4%, 95.2% and 93.2%, respectively. Conclusions:The mGI-RADS has a high clinical value in the differential diagnosis of ovarian tumors.

Objective To investigate the incidence of abnormalities and adverse reactions of intrauterine device ( IUD) by three‐dimensional transvaginal ultrasound ,and guide the correct selection of IUD . Methods Female volunteers who came to our hospital for health check‐up from July 2016 to February 2018 were selected as subjects to obtain information of the belt loop time and number of loops . T hree‐dimensional transvaginal ultrasound was used to observe and record the IUD types and positions . T he incidence of location abnormalities ,adverse reactions and cervical lesions of differnet types of IUD were retrospectively analyzed . Results T wo thousand one hundred and thirteen eligible subjects were included in the study ,including 423 cases of McuIUD ,506 cases of circular ring ,405 cases of uterine ring ,372 cases of T‐ring and 407 cases of γ ring . T he statistical analysis showed that the incidence of adverse reactions after wearing rings was as high as 34 .07% ,and the adverse reaction rate of different IUD was different . McuIUD had the highest incidence of adverse reactions ,compared with the circular ring ,the uterine ring ,the T‐ring , and the γ ring ( 47 .99% vs 25 .88% ,27 .41% ,37 .10% ,33 .66% ; all P ＜ 0 .05 ) . Different IUD had different incidence of adverse reactions . T he low back pain rate in McuIUD and γ ring were higher than the other 3 IUDs . T he low back pain rate of McuIUD was higher than the circular ring ,the uterine ring and the T‐ring ( 23 .4% vs 15 .71% ,14 .07% ,13 .7 1% ; all P ＜ 0 .01 ) . Different types of IUD had different incidence of location anomalies . T he descending rate of the circular ring was higher than that of the McuIUD and γ ring ( 6 .32% vs 3 .30% ,3 .19% ; all P＜ 0 .05 ) ,and no statistically significant difference compared with the uterine ring and T‐ring ( 6 .32% vs 3 .45% ,4 .30% ; all P ＞0 .05 ) . T he incarceration rate of the mother ring was higher than that of the circular ring ,uterine ring and T‐ring ( 3 .07% vs 0 ,0 .25% ,0 .54% ;all P ＜0 .01) . T he intrauterine rotation rate of uterine ring and γ ring were higher than that of McuIUD , circular ring and T‐ring ( all P ＜0 .01) . Pregnancy ＞1 and ring times ≥2 were related with the occurrence of adverse reactions of IUD ( P ＜ 0 .05 ) . T he incidence of cervical erosion in the T‐ring was the highest compared with the McuIUD ,circular ring ,uterine ring ,and γ ring ( 30 .65% vs 16 .78% ,16 .80% ,18 .02% , 17 .69% ; all P ＜0 .01) . Conclusions The incidence of adverse reactions is higher in women with different types of IUD . T he incidence of different types of abnormal IUD position is different . T he frequency of pregnancy and band ring are related to the occurence of adverse veactions to IUD .

Rhizoma Corydalis ,a herb with analgesic effects and being able to promote blood circulation ,has been used for more than hundreds of years in China .Rhizoma Corydalis mainly comes from Zhejiang ,Jiangsu ,Hubei and Hunan prov-ince .The major effective components are alkaloids ,which have sedative ,hypnotic ,analgesic pharmacological effects .It has in-dications for chest ,rib and back pain ,dysmenorrhea ,traumatic injury ,etc .The main formulation of Rhizoma Corydalis is vinegar product .The processing technology has significant effect on the content of active components and therapeutic efficacy . It is very important to improve and optimize decoction method .This paper reviewed the processing technology and the pharma-cological effects of Rhizoma corydalis .

Objective:To optimize the formula of film-forming materials for compound rhizoma corydalis plastics. Methods:Poly-vinyl alcohol 124 (PVA124) and bletilla striata gum were selected as the film-forming materials. The amounts of PVA 124, bletilla striata gum, glycerol and absolute ethanol were used as the influencing factors in an orthogonal test with the composite score of film-forming time and appearance as the indices to screen out the best composition of film-forming materials. Results:The best composition of film-forming materials was as follows:8 g PVA 124, 1 g bletilla striata gum, 2 ml glycerol and 15 ml ethanol. Conclusion:The op-timized film-forming materials can be used for the preparation of compound rhizoma corydalis plastics.

Objective Compared with povidone iodine solution to clean,glutaraldehyde immersion,highpressure steam sterilization of three disinfection methods for dental handpiece sterilization effect.Methods Select the 20 dental clinical used over phone,were randomly divided into A group,B group,C group,D group 5,A group for the control group,only cleaning method did not use any disinfection,B group was 1% Polyvinylpyrrolidone iodine solution,wipe,C group were soaked in 2% glutaraldehyde soluton,D group were treated with high-pressure steam sterilization,after each phone were inoculated with bacteria sample dish,the more monitoring of four groups of bacterial culture.Results A group of bacterial culture for the intensive growth of bacteria,B group had bacterial growth for the(+ ～+ + +),bacterial growth;C group had bacterial culturegrowth was(+ ～ + +),bacterial growth;D group of bacterial culture Growth of(-),no bacterial growth.Conclusion High-pressure steam sterilization was the disinfection of dental handpieces most effective way.