AIM: To investigate the mechanism of elemene synergistic bortezomib against multiple myeloma based on ROS-NF-κB-p38MAPK signaling pathway. METHODS: CCK-8 assay was used to detect cell activity. Nude mice were randomly divided into control group, bortezomib (BTZ) group, elemene (ELE) group and combination group. Each group was treated with BTZ, ELE and BTZ combined with ELE, respectively. Tunel staining was performed to observe the apoptosis of tumor tissues. The expressions of Caspase-3, Bcl-2, NF-κB and p38 MAPK were detected by Western Blot. Cell cycle, apoptosis and reactive oxygen species (ROS) expression were detected by flow cytometry using human myeloma U266 cells. RESULTS: When 4.0 μmol/L ELE combined with 50 nmol/L BTZ treated U266, the cell activity was significantly reduced compared with that of NC, BTZ and ELE groups (P< 0.05). The tumor volume of nude mice in BTZ group, ELE group and combined group was significantly reduced compared with the control group (P <0.05), and the combined group was the smallest. Tunel staining results showed that the apoptosis level in the control group was lower than that in the BTZ group, ELE group and the combined group (P<0.05), and the combined group had the lowest apoptosis level. Compared with the control group, the expressions of Caspase-3 and p38 MAPK in BTZ group, ELE group and combination group were significantly increased, while the expression of Bcl-2 was significantly decreased. The apoptosis level and expression of ROS in BTZ group, ELE group and the combined group was significantly increased compared with the control group (P<0.05). CONCLUSION: ELE can enhance the role of BTZ in promoting apoptosis of myeloma cells, which may be achieved by regulating ROS/NF-κB/p38 MAPK signaling pathway to enhance the level of apoptosis of tumor cells to achieve anti-tumor effect.

Objective:To investigate the efficacy of laparoscopic sleeve gastrectomy (LSG) in morbidly obese patients aged 10 to 21 years.Methods:We conducted a retrospective analysis of clinical data from 89 out of 200 patients who underwent LSG at the Gastrointestinal Surgery/Weight Loss Center of the First Affiliated Hospital of Jinan University between January 2015 and December 2020. The primary outcome measures were the completion rate of LSG, the incidence of perioperative complications, and weight-related indicators 3, 6, 12, and ≥24 months postoperatively. Additionally, we compared glucose metabolism, lipid metabolism, vitamin levels, liver function, and other relevant biochemical variables before and after surgery. Normally distributed continuous data are presented as x±s. Because the numbers of patients at each follow-up time point were not identical with the number of patients in the study cohort preoperatively, independent sample t-tests were used for intergroup comparisons. Non-normally distributed continuous data are presented as M( Q1, Q3), and Mann-Whitney U tests were used for intergroup comparisons. Results:Among the 89 patients, 35 were male (39.3%), the mean age was (18±2) years, and mean body mass index (BMI) 38.5±4.8 kg/m2; 37 of the patients having a BMI greater than 40 kg/m2. Additionally, 63 patients (70.8%) had fatty livers, 34 (38.2%) hyperuricemia, 31(34.8%) sleep apnea syndrome, 20 (22.4%) gastroesophageal reflux, eight (8.9%) type 2 diabetes, and two (2.2%) hypertension. All 89 patients underwent LSG surgery successfully, with no conversions to open surgery. During the perioperative period, there were no cases of major bleeding, gastric leakage, or infections. Notable postoperative symptoms included nausea, vomiting, and pain, most of which improved by the second postoperative day. BMI values 3, 6, and 12 months postoperatively had decreased to 31.5±5.8 kg/m2, 28.6±4.3 kg/m2, and 26.3±4.4 kg/m2, respectively. All of these BMI values differed significantly from preoperative values (all P<0.05). At 12 and ≥24 months postoperatively, the percentages of total weight loss were (31.3±9.3)% and (33.1±10.5)%, respectively, both differing significantly from 3 months postoperatively (20.5±5.1)% (all P<0.05). The percentages of excess weight loss at 12 and ≥24 months postoperatively were 91% (70%, 113%) and 95% (74%, 118%) , respectively, both differing significantly from the percentage of total weight loss 3 months postoperatively (56% [45%, 72%]) (both P<0.05). Alanine transaminase and aspartate transaminase serum concentrations decreased from preoperative values of 44.4 (25.5, 100.5) U/L and 29.0 (9.5, 48.0) U/L to 14.0 (10.8, 18.3) U/L and 13.0 (10.5, 17.3) U/L, respectively, ≥24 months postoperatively. Hemoglobin A1c decreased from 5.6 (5.3, 5.8)% preoperatively to ≥24 months postoperatively 5.3 (5.0, 5.4)%. High-density lipoprotein increased from 1.0 (0.9, 1.2) mmol/L preoperatively to 1.4 (1.1, 1.6) mmol/L ≥24 months postoperatively. Vitamin B12 decreased from 350.0 (256.8, 441.3) μg/L preoperative to 230.3(195.4, 263.9) μg/L ≥24 months postoperatively. All differed significantly from preoperative values (all P<0.05). Conclusion:LSG has favorable efficacy in morbidly obese patients aged 10 to 21 years. However, further confirmation is required through long-term, multicenter, randomized, controlled trials.

Objective:To investigate the efficacy of laparoscopic sleeve gastrectomy (LSG) in morbidly obese patients aged 10 to 21 years.Methods:We conducted a retrospective analysis of clinical data from 89 out of 200 patients who underwent LSG at the Gastrointestinal Surgery/Weight Loss Center of the First Affiliated Hospital of Jinan University between January 2015 and December 2020. The primary outcome measures were the completion rate of LSG, the incidence of perioperative complications, and weight-related indicators 3, 6, 12, and ≥24 months postoperatively. Additionally, we compared glucose metabolism, lipid metabolism, vitamin levels, liver function, and other relevant biochemical variables before and after surgery. Normally distributed continuous data are presented as x±s. Because the numbers of patients at each follow-up time point were not identical with the number of patients in the study cohort preoperatively, independent sample t-tests were used for intergroup comparisons. Non-normally distributed continuous data are presented as M( Q1, Q3), and Mann-Whitney U tests were used for intergroup comparisons. Results:Among the 89 patients, 35 were male (39.3%), the mean age was (18±2) years, and mean body mass index (BMI) 38.5±4.8 kg/m2; 37 of the patients having a BMI greater than 40 kg/m2. Additionally, 63 patients (70.8%) had fatty livers, 34 (38.2%) hyperuricemia, 31(34.8%) sleep apnea syndrome, 20 (22.4%) gastroesophageal reflux, eight (8.9%) type 2 diabetes, and two (2.2%) hypertension. All 89 patients underwent LSG surgery successfully, with no conversions to open surgery. During the perioperative period, there were no cases of major bleeding, gastric leakage, or infections. Notable postoperative symptoms included nausea, vomiting, and pain, most of which improved by the second postoperative day. BMI values 3, 6, and 12 months postoperatively had decreased to 31.5±5.8 kg/m2, 28.6±4.3 kg/m2, and 26.3±4.4 kg/m2, respectively. All of these BMI values differed significantly from preoperative values (all P<0.05). At 12 and ≥24 months postoperatively, the percentages of total weight loss were (31.3±9.3)% and (33.1±10.5)%, respectively, both differing significantly from 3 months postoperatively (20.5±5.1)% (all P<0.05). The percentages of excess weight loss at 12 and ≥24 months postoperatively were 91% (70%, 113%) and 95% (74%, 118%) , respectively, both differing significantly from the percentage of total weight loss 3 months postoperatively (56% [45%, 72%]) (both P<0.05). Alanine transaminase and aspartate transaminase serum concentrations decreased from preoperative values of 44.4 (25.5, 100.5) U/L and 29.0 (9.5, 48.0) U/L to 14.0 (10.8, 18.3) U/L and 13.0 (10.5, 17.3) U/L, respectively, ≥24 months postoperatively. Hemoglobin A1c decreased from 5.6 (5.3, 5.8)% preoperatively to ≥24 months postoperatively 5.3 (5.0, 5.4)%. High-density lipoprotein increased from 1.0 (0.9, 1.2) mmol/L preoperatively to 1.4 (1.1, 1.6) mmol/L ≥24 months postoperatively. Vitamin B12 decreased from 350.0 (256.8, 441.3) μg/L preoperative to 230.3(195.4, 263.9) μg/L ≥24 months postoperatively. All differed significantly from preoperative values (all P<0.05). Conclusion:LSG has favorable efficacy in morbidly obese patients aged 10 to 21 years. However, further confirmation is required through long-term, multicenter, randomized, controlled trials.

Objective To analyze the angiographic results of intracranial aneurysms without complete embolization immediately after low-profile visualized intraluminal support device (LVIS) stent at mid-term follow up,and explore the risk factors for healing of partial occluded aneurysms.Methods One hundred and sixty-one patients with intracranial aneurysms treated by LVIS stent embolization in our hospital from December 2014 to December 2018 were selected;193 aneurysms in total,including 93 un-ruptured aneurysms and 100 ruptured aneurysms,were noted.DSA was performed immediately after operation to evaluate the degrees of aneurysm embolization according to Raymond grading criteria.The embolization degrees of aneurysms were compared at 8 months after surgery and immediately after surgery,and the healing of aneurysms (Raymond grading 1) was calculated.Univariate Logistic regression analysis and multivariate Logistic regression analysis (forward maximum likelihood ratio method) were used to screen the risk factors for healing of incomplete aneurysm embolization.Results The embolization degrees immediately after surgery were as follows:78 were with Raymond grading 1,54 with Raymond grading 2,and 61 with Raymond grading 3;complete aneurysm embolization were noted in 78 aneurysms (40.41％) and incomplete aneurysm embolization were noted in 115 (59.59％).Follow up (8 months after surgery) results indicated that,of the 193 aneurysms,171 were with Raymond grading 1,10 with Raymond grading 2,and 12 with Raymond grading 3;there were 171 aneurysms (88.60％) having complete aneurysm embolization and 22 (11.40％) having incomplete aneurysm embolization.In the aneurysms having incomplete aneurysm embolization immediately after surgery,the healing rate was 81.74％ (94/115).Univariate Logistic regression analysis showed that hypertension,diabetes mellitus,posterior circulation aneurysm,dissecting aneurysm,body-neck ratio and embolization degrees immediately after surgery were risk factors for healing of aneurysms (P＜0.05).Multivariate Logistic regression analysis revealed that posterior circulation aneurysm and diabetes mellitus were independent risk factors for healing of aneurysms (P＜0.05).The area under the curve of receiver operating characteristic curve of the regression model was 0.755,indicating that the prediction efficiency of the regression model was moderate.Conclusion Treatment of intracranial aneurysms with LVIS stent is effective,and the complete occlusion rate is high,even in the aneurysms with incomplete embolization immediately after surgery;aneurysms located in the posterior circulation and associated with diabetes can affect the treatment of aneurysms.

Objective To analyze the residents′ willingness to hierarchical medical system and their intention of first visit hospitals.Methods Customized questionnaire was used for in-home survey of 1 500 households in three cities of Shandong province,with 773 of the residents aware of hierarchical medical system used as the study samples.Descriptive analysis and multi factor Logistic regression were employed for statistical analysis of their intention.Results 85.0% the residents expressed willingness to embrace hierarchical medical system.55.0% of the residents prefer primary medical institutions as their first visit in case of mild illness,a choice chosen by more rural residents(58.9%)than urban residents(48.0%),a difference of statistical significance(P<0.01).In the case of severe illness,87.3% of them prefer major hospitals as their first visit,and more urban residents(94.3%)take this option than the rural residents(83.3%),a difference of statistical significance(P<0.01).Service ability of medical institutions and residents′ concept for medical service played a key role in their choice of first visit hospitals.Conclusions The key to ensuring the implementation effect of hierarchical medical system lies in better service ability of primary medical institutions and correction of residents′ irrational medication concepts.

Objective To analyze the correlation factors that promote or impede the residents to preferentially use essential medicines.Methods Adopting stratified random sampling method,1 700 households selected from 5 cities of Shandong province were investigated with a questionnaire.The framework of Andersen behavior model of health service utilization was used as the framework,with such methods as descriptive analysis and univariate logistic regression models for the analysis and evaluation of relevant information.Results The residents′ tendency factor,ability factor and environmental factor influence their preference to use essential medicines,while the requirement factor plays a minimal role.There was a significant difference for the preference of combined medication,first visit preference and self-medication experience,the efficacy and policy response of essential medicine from logistic regression analysis.Conclusion At present,the government should focus on the construction of the formation mechanism of the residents′drug use behavior and the policy response mechanism of essential medicine system.

Objective To investigate the role of ORAI1 in the balloon injury-induced rat carotid artery neointimal formation and the relationship between ORAI1 and sodium and calcium exchanger(NCX) 1.Methods According to condition of carotid artery balloon injury and observation time,Sprague Dawley rats were divided into 3 groups including sham group,7 days injury group,and 14 day injury group (n =3 each).According to virus of transfection,Sprague Dawley rats were divided into 2 groups including negative control group (negative lentiviruses particles transfection after establishment of carotid artery balloon injury model,n =3) and siORAI1 group(siORAI1 transfection after operation,n =3).Carotid artery neointimal formation was analyzed after HE staining.The ORAI mRNA expression level was detected using real-time PCR.The proliferating cell nuclear antigen (PCNA),ORAI and NCX1 protein expression level were measured by Western blot.Results (1) Carotid artery intima was significantly thicker in 7 days and 14 days injury group than in sham group((0.54 ±0.11) μm2 and (0.89 ±0.12) μm2 vs.(0.11 ±0.08) μm2,both P ＜0.05).Relative expression level of PCNA protein was significantly higher in 7 days and 14 days injury group than in sham group(1.43 ±0.16 and 1.95 ±0.16 vs.1,both P ＜0.05).Relative mRNA expression level of ORAI1 (1.39 ± 0.14 and 1.78 ± 0.21 vs.0.56 ± 0.09,both P ＜ 0.05) and protein expression level (1.42 ± 0.19 and 1.78 ±0.22 vs.1,both P ＜0.05) were significantly higher in 7 days and 14 days injury group than in sham group.(2) After 14 days,relative expression level of ORAI1 protein was significantly lower in siORAI1 group than in negative control group (0.21 ±0.16 比 1,P ＜0.05).Carotid artery intima thickness was significantly reduced in siORAI1 group compared to negative control group((0.19 ± 0.14)μm2 vs.(0.91 ±0.23) μm2,P＜0.05).Relative expression level of NCX1 protein was also significantly lower in siORAI1 group than in negative control group(0.53 ±0.13 比 1,P ＜0.05).Conclusions ORAI1 may play a key role in the balloon injury-induced neointimal formation and vascular smooth muscle cells proliferation in rat carotid artery.ORAI1 knockdown could down regulate the NCX1 expression and attenuate the balloon injury-induced neointimal formation and vascular smooth muscle cells proliferation in rat carotid artery.

Objective:To establish a method based on the iPLEX analysis of MassARRAY mass spectrometry platform to detect multiplex genetic mutations among Chinese lung cancer patients. Methods:We reviewed the related literature and data of lung cancer treatments. We also determined 99 mutation hot spots in 13 target genes, namely, EGFR, KRAS, ALK, FGFR1, FGFR2, FGFR3, PIK3CA, BRAF, PTEN, MET, ERBB2, AKT1, and STK11, which are closely related to the pathogenesis, drug resistance, and metastasis of lung cancer and are associated with relevant transduction pathways. A total of 297 primers comprising 99 paired forward and reverse amplification primers and 99 matched extension primers were designed by using Assay Design in accordance with the mutation label and format requirements of the MassARRAY platform. The detection method was established by analyzing eight cell lines and six lung cancer specimens;the proposed method was then validated through comparisons with a LungCarta kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases. Results:The proposed method could detect multiplex genetic mutations in the lung cancer cell lines, and this finding is consistent with that observed using previously reported methods. The proposed method could also detect such mutations in clinical lung cancer specimens;this result is also consistent with that observed by using the LungCarta kit. However, an FGFR2 mutation was detected only by using the proposed method. The measured sensitivity and specificity were 100%and 96.3%, respectively. Conclusion:The proposed MassARRAY technology-based method could detect multiplex genetic mutations among Chinese lung cancer patients. Indeed, the proposed method can be potentially applied to detect mutations in cancer cells.

ObjectiveTo investigate the fusion sequence complexity of EML4-ALK in non-small cell lung cancer (NSCLC) patients,and the potential mutation in tyrosine kinase ( TK ) domain of ALK gene.MethodsIn routine practice,a novel echinoderm microtubule-associated protein-like4 and anaplastic lymphoma kinase (EML4-ALK) V3c variant was detected by rapid amplification of cDNA ends-polymerase chain reaction ( RACE-PCR )-sequencing technology in a patient with NSCLC.The further consecutive 39 cases( total of 40 cases)were screened by use of reverse transcription (RT)-PCR for EML4-ALK fusion.Positive PCR products were purified and cloned into T vectors,transformed into DH5a germ cells and colony picked up and sequenced for sequence complexity analysis.Tyrosine kinase domain of ALK was amplified by RT-PCR and sequenced.ResultsThree out of 40 cases had EML4-ALK fusion.One case had six novel variants of EML4-ALK co-existing,termed as V3c ( 64.6％ ),V3d ( 25.0％ ),V3e ( 2.1％ ),V3f (4.2％ ),V3g(2.1％ )and V3h(2.1％ ) variants,whereas without common V3a and V3b variants.In other two positive cases,one was V1 variant,another was concurrent V2,V3a and V3b variants.No mutations were detected in the TK domain of EML4-ALK in any case.ConclusionsSeveral EML-ALK variants could co-exist in a given lung cancer tissue,which suggest that the diversity and sequence complexity of EML4-ALK fusion are exist.Attentions should be paid to screen all the variants in clinic to improve the pick-up rate.

Objective To establish a HRM assay to screen for KRAS mutations in clinical colorectal cancer patients.Methods The sensitivity of HRM was analyzed by detecting somatic mutations in exon 2,notably codons 12 and 13 of the KRAS gene in the serial plasmid mixture samples which were mixed using the different proportions mutation plasmid and wide type plasmid of KRAS.HRM analysis was performed for KRAS on DNA insolated from a panel of 60 colorectal cancer samples derived from fresh tissues.The results were compared with the direct sequencing data.Results After the PCR amplification,the mutation results could be available by performing HRM analysis in the same tube on a real time PCR machine with HRM capability.HRM detection could identify KRAS mutation in a proportion of 10% of mutation plasmid DNA.All 60 samples identified the KRAS mutation by HRM and sequencing.17 samples were positive(28.3%) by HRM for KRAS exon 2 mutations,and 15 samples were confirmed the presence of codon 12 or 13 mutations(25.0%) and the other 2 samples were wild type by sequencing.The 60 samples detected by HRM were given 100% sensitivity with 96% specificity.Conclusions HRM is a sensitive intube methodology to screen for mutations in clinical samples.HRM will enable high-throughput screening to gene mutations to allow appropriate therapeutic choices for patients and accelerate research aimed at identifying novel mutations in human cancer.