OBJECTIVE To investigate the relationship of long non-coding RNA （LncRNA） metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） and doxorubicin （DOX） resistance in osteosarcoma （OS） cells. METHODS MG-63 and MG-63/DOX cells were treated with different concentrations of DOX （0， 0.01， 0.05， 0.1， 1 μmol/L）， and survival rates and half maximal inhibitory concentration were determined using CCK-8 assay. The expressions of LncRNA MALAT1 in MG-63 and MG-63/ DOX cells were detected by real-time quantitative fluorescence PCR. MG-63/DOX cells were divided into Control group， knocking down LncRNA MALAT1 negative control （sh-NC） group， sh-MALAT1 group， sh-MALAT1+anti-NC group， and sh-MALAT1+ anti-miR-154-5p group. The expressions of LncRNA MALAT1， miR-154-5p and cyclin D1 （CCND1） mRNA in MG-63/DOX cells of each group were detected. The effects of knocking down LncRNA MALAT1 on the proliferation， migration， invasion， and apoptosis of MG-63/DOX cells were detected by CCK-8 assay， scratch test， Transwell experiment and flow cytometry， respectively. The expression of proliferating cell nuclear protein （PCNA） and CCND1 protein in MG-63/DOX cells was detected by Western blot assay. Interactions between LncRNA MALAT1 and miR-154-5p， miR-154-5p and CCND1 were detected by dual luciferase reporter gene experiment. RESULTS Compared with 0 μmol/L DOX， 0.01， 0.05， 0.1 and 1 μmol/L DOX could reduce the survival rates of MG-63 and MG-63/DOX cells （except for 0.01 μmol/L DOX） （P＜0.05）， IC50 were 0.07 and 0.13 μmol/L， respectively. The survival rate， cell migration number and invasion number of MG-63/DOX cells， scratch closure rate， mRNA expressions of LncRNA MALAT1， mRNA and protein expressions of CCND1， and PCNA protein expression in sh-MALAT1 group were significantly lower than sh-NC group and Control group； the apoptosis rate and miR-154-5p expression were significantly higher than sh-NC group and Control group （P＜0.05）. sh-MALAT1+anti-miR-154-5p group was able to reverse the aforementioned biological effects in sh-MALAT1 group （P＜0.05）. In MG-63/DOX cells transfected with both MALAT1-wild type （WT） and CCND1-WT， the luciferase activity in the miR-154-5p mimic group was significantly lower than mimic negative control group （P＜ 0.05）. CONCLUSIONS Knocking down LncRNA MALAT1 can inhibit the DOX resistance of OS cells， and its mechanism may be targeting the miR-154-5p/CCND1 axis.

Objective:To explore the severe complications, such as blindness, brain infarction and even death caused by cosmetic autologous fat filler injection and their underlying mechanisms.Methods:From May 2022 to October 2023, 64 male New Zealand rabbits were selected in the clinical laboratory of Shaanxi University of Chinese Medicine and divided into 8 groups with 8 rabbits in each group. They were divided into grinding fat group, fat granule group, fat lipid group and normal saline group, and each group was further divided into 0.2 ml group and 0.4 ml group. Fat was cut from the rabbit groin, then chopped or treated with collagenase I, and centrifuged to separate fat lipids and fat particles, as well as other tissues. The rabbit facial artery was exposed along the incision below the mandibular angle, and 0.2 or 0.4 ml of chopped fat, fat particles, and lipids were retrogradely injected into the facial artery in each group, and then the incision was closed under the microscope. Ophthalmic and neurological symptoms were observed and recorded after surgery, and visual electrophysiology and fundus microscopy were performed to verify visual acuity and fundus artery obstruction.Results:Two weeks after surgery, the incidence of ophthalmic symptoms in the 0.2 ml injection group was 100% in the grinding fat group, 62.5% in the fat granule group, 0 in the lipid group and 0 in the normal saline group. The complication rates of 0.4 ml embolic injection were 100%, 87.50%, 12.5% and 0, respectively. The incidence of neurological symptoms was 62.5%, 25.0%, 0 and 0, respectively. Mortality rates were 37.5%, 12.5%, 0 and 0 after injection of 0.2 ml, and 100%, 50%, 0 and 0 after injection of 0.4 ml, respectively.Conclusions:Animal models have shown that grinding adipose tissue without collagenase I treatment is more likely to cause serious complications, simple lipid entry into blood vessels does not block relevant arteries, and fat volume is positively correlated with the incidence of postoperative complications.

Objective:To construct pancreatic cancer cell lines expressing luciferase and mesothelin (MSLN), and evaluate the feasibility of using them as target cells in analyzing the cytotoxicity activity of immune cells.Methods:Lentiviral vectors expressing luciferase and MSLN genes were constructed, and pancreatic cancer cell lines were infected after lentivirus packaging. Single-cell clones were obtained by limited dilution following antibiotic screening, and the stable expression of the target genes were verified. These cells were used as target cells to detect the cytotoxicity of immune cells by real-time cell analysis (RTCA) and luciferase activity. Besides, these luciferase-expressing cells were transplanted into B-NDG mice to establish the animal models of pancreatic cancer, and in vivo optical imaging technology was used to detect the expression of luciferase and monitor the tumor growth in mice. The cytotoxicity of chimeric antigen receptor T (CAR-T) cells was verified in these animal models. Results:Three pancreatic cancer cell lines, panc-1-luc, panc-1-luc-MSLN and capan-2-luc, that could stably express luciferase and MSLN genes were successfully constructed. The expression of the reporter gene in these cells were high, and positively correlated with the number of cells. There were 95.6% of panc-1-luc-MSLN cells expressing MSLN. MSLN-CAR-T cells had specific killing effect on MSLN-positive panc-1-luc-MSLN cells and capan-2-luc cells, with the minimum killing rates of (70.00±18.19)% and (57.00±5.29)%, respectively. But they had no cytotoxicity to MSLN-negative panc-1-luc cells. RTCA results showed that MSLN-CAR-T cells were able to lyse all three pancreatic cancer cell lines, and the minimum killing rates were (56.33±7.64)%, (93.00±2.65)% and (26.33±28.15)%, respectively. The killing of target cells by NK-92MI cells was not depended on MSLN expression. The cytotoxicity in the mice models of pancreatic cancer was consistent with the results in vitro. The in vivo and in vitro test results suggested that the expression of luciferase by target cells could reflect the cytotoxicity of immune cells. Conclusions:This study establishes three pancreatic cancer cell lines stably expressing luciferase, which can be used to evaluate the cytotoxicity of immunotherapy products targeting tumor cells in vitro and in vivo.

Proto-Oncogene Proteins c-akt/metabolism* ; Butyrates ; Ligands ; Glucose ; Receptors, G-Protein-Coupled/metabolism* ; Glycogen Synthase Kinases/metabolism* ; Glycogen Synthase Kinase 3 beta ; Phosphorylation

Abstract OBJECTIVE To develop LC-MS method for the simultaneous determination of impurities A, C, and D of caspofungin acetate. METHODS Waters CORTECS® C18+(4.6 mm×150 mm, 2.7 μm) was used as the chromatography column. Mobile phase A and B were 0.1% formic acid-H2O and 0.1% formic acid-CH3CN, respectively. Electrospray ion source-single quadrupole mass spectrometry was used to detect impurities A and C in positive ion mode and impurity D in negative ion mode. RESULTS The correlation coefficient r was ≥ 0.999 in linearity ranges of impurities A, C and D. The average recoveries were 100.5%, 104.1% and 105.2%, respectively, with RSD<4%(n=6). The LOQs (S/N=10) of impurities A, C and D were 31.8, 6.99 and 15.5 ng·mL-1 respectively. The contents of impurities A, C and D in the three samples were all below the limits. CONCLUSION The developed LC-MS method is simple, sensitive, and applicable, which can be used to simultaneously determine impurities A, C and D in caspofungin acetate and can also provide a reference for the detection of other impurities in caspofungin acetate.

OBJECTIVE To analyze caspofungin acetate and the samples under different strong degradation conditions by LC-QTOF-MS, and to study the characteristics in mass spectra of caspofungin and its related impurities(impurities A, B, C, D and E). METHODS Chromatographic separation was accomplished on Waters CORTECS® C18+(4.6 mm×150 mm, 2.7 μm) column using a gradient elution with monile phase of 0.1% formic acid-H2O(A) and 0.1% formic acid-CH3CN(B) at a flow velocity of 0.6 mL·min-1; The analytes was detected in positive ion scan mode by ESI-QTOF-MS. RESULTS In MS1 spectra, except that impurity D mainly showed single-charge quasi-molecular ion, caspofungin and the other four impurities showed muti-charge quasi-molecular ions with high abundance; In MS2 spectra, caspofungin and its impurities that containing ethylenediamine generated fragment ions at m/z 1 033 by losing the ethylenediamine and the groups attached to it; caspofungin and its impurities produced a series of fragment ions mainly through the cleavage of peptide bonds, as well as through the loss of hydroxyl, acyl, or amino groups from amino acid residues; Impurity A and C showed characteristic fragment ions m/z 137.070 8 and m/z 77.071 1 with high abundance, respectively, which could be used to distinguish them from caspofungin and the other impurities. CONCLUSION Caspofungin and its five impurities have distict characteristics in their mass spectra. The research results can provide reference for identifying the structures of unknown impurities that may occur in the production process of caspofungin, so as to quickly discover the potential problems in the production process and reduce the quality risk of the products.

Objective: To identify the lifestyle pattern of adolescents in Guangzhou and to provide a scientific basis for targeted and individualized interventions for adolescents with different lifestyle pattern. Methods: A survey was conducted by questionnaire among 12 540 students from 27 middle schools in the urban area of Guangzhou using a convenient sampling method. The latent class model (LCM) was used to identify lifestyle patterns of adolescents, and further analyses were conducted to compare differences in dietary habits and willingness to exercise among the different classes. Results: Three lifestyle patterns were identified based on the latent class model: "high sweet snacks/excessive screen time, sleep and exercise deficiency group", 3 797 people, accounting for 30.3%; "low nutrition diet/severe sleep and exercise deficiency group", 2 745 people, accounting for 21.9%; "general diet/sleep and exercise deficiency group", 5 998 people, accounting for 47.8%. Adolescents of different classes had different perceptions of their eating habits and different degrees of willingness to participate in physical exercise, these differences were statistically significant(χ 2=671.54, Z=153.16, P<0.05). Conclusion The results of the latent class model showed that the three classes of adolescents had their own unique characteristics. It is necessary to inform them of their needs, provide guidance and implement targeted interventions according to the unique characteristics of the different lifestyle patterns in empirical work.

Objective: To understand the knowledge, attitudes and practices of drinking water among college students of Hebei Province, to provide evidence for health education. Methods: A total of 715 subjects were selected from a university in Baoding City, Hebei Province in March 2017 by random cluster sampling. Knowledge, attitude and behavior of drinking water among college students were investigated by self-designed questionnaires. Results: The awareness rate of water consumption of 1 500-1 700 mL was 19.0%, the awareness rate of drinking modes was 95.0%, the awareness rate of suitable drinking time was 30.1%, the awareness rate on drinking water was higher among girls and medical major sutdents (χ2=22.16, 12.49, P<0.05); the awareness rate on the relationship between insufficient drinking water and stroke, hypertension and coronary heart disease was generally low, with sophornore higher than freshman (χ2=11.10, P<0.05). The most common medium for accessing drinking water knowledge among the respondents was network (60.0%), followed by campus publicity/classroom teaching (48.3%). 99.6% of the respondents believed that drinking water was important to health, 98.3% were willing to change their drinking habits, 93.0% were willing to pay attention to drinking water education, and the proportion of female students was higher than that of male students (χ2=5.46, 6.49, 8.01, P<0.05). In real life, 48.4% of the respondents drank water on an empty stomach in the morning and 40.7% before going to bed. 45.5% of the respondents drank water when they thought of it. Conclusion College students in Hebei province are generally lack of appropriate awareness on water drinking, which varied in sex, grade and major. While the attitude towards water drinking is satisfactory, drinking behavior need to be improved.

Objective:To investigate the effect of postoperative hypoalbuminemia on acute kidney injury (AKI) after cardiac surgery under cardiopulmonary bypass (CPB).Methods:The clinical data of adult patients undergoing cardiac surgery under CPB were retrospectively analyzed. The difference between preoperative and postoperative serum albumin level was compared. The patients were divided into hypoalbuminemia group (≤35 g/L) and non-hypoalbuminemia group (>35 g/L) according to the lowest serum albumin concentration within 48 hours after surgery. The incidence and severity of postoperative AKI were compared between the two groups. Univariate analysis and binary logistic regression analysis were used to evaluate the effect of postoperative hypoalbuminemia on the incidence of postoperative AKI.Results:Among the 749 patients, the serum albumin level after cardiac surgery was significantly lower than that before surgery ( Z=-15.739, P<0.001), and the proportion of patients with hypoalbuminemia increased from 9.6% to 27.6% ( χ2=83.516, P<0.001). Postoperative AKI occurred in 273 patients, including 109 cases (52.7%) in hypoalbuminemia group and 164 cases (30.3%) in non-hypoalbuminemia group. The incidence of AKI in hypoalbuminemia group was significantly higher than that in non-hypoalbuminemia group ( χ2=32.443, P<0.001), and the severity of AKI in hypoalbuminemia group increased than that in non-hypoalbuminemia group ( Z=-2.098, P=0.036), and the time of hospital stay extended ( Z=-2.442, P=0.015). After adjusted by gender, age, preoperative hypoalbuminemia, comorbidities (hypertension, hyperuricemia, diabetes mellitus, cerebrovascular disease), renal insufficiency, preoperative heart function, coronary angiography, CPB time, aorta blocking time, type of heart surgery and postoperative hypotension, binary logistic regression analysis revealed that postoperative hypoalbuminemia was an independent risk factor for CPB-associated AKI ( OR=2.319, 95% CI 1.586-3.392, P<0.001). Conclusions:AKI is a common complication following cardiac surgery under CPB. Serum albumin after CBP is significantly lower than that before CBP, and postoperative hypoalbuminemia within 48 hours after surgery is an independent risk factor for AKI.

Objective: To examine the association of pre-pregnancy body mass and weight gain during pregnancy with macrosomia. Methods: From January 2015 to December 2015, a total of 20 477 pregnant women were recruited by probabilistic proportional scale sampling with simple randomization in Sichuan, Yunnan and Guizhou Provinces. Basic information of pregnant women, weight gain during pregnancy and weight of newborn were collected. A multiple logistic regression model was used to assess the association between the pre-pregnancy body mass and gestational weight gain indicators with macrosomia. Results: 20 321 mother-infant were included in the final analysis. 20 321 pregnant women were (30.09±4.10) years old and delivered at (39.20±1.29) weeks, among which 12 341 (60.73%) cases were cesarean delivery. The birth weight of 20 321 infants were (3 292.26±431.67) grams, and 970 (4.77%) were macrosomia. The multiple logistic regression model showed that after adjusting for the age of women, compared to the normal weight group in the pre-pregnancy, the overweight and obesity group elevated the risk of macrosomia, with OR (95%CI) about 1.99 (95%CI: 1.69−2.35) and 4.05 (95%CI: 3.05−5.39), respectively. After adjusting for the age, the pre-pregnancy BMI, delivery weeks, delivery mode and infant′s gender, compared to the weight-gain appropriate group, higher weight gain rate in the mid-pregnancy and excessive total gestational weight gain elevated the risk of macrosomia, with OR (95%CI) about 1.99 (95%CI: 1.66−2.39) and 1.80 (95%CI: 1.55−2.08), respectively. Conclusion The overweight before pregnancy, obesity before pregnancy, the rate of weight gain in the second trimester and the high total weight gain during pregnancy could increase the risk of macrosomia.