[Effect of SLC7A11 gene downregulation on the gefitinib resistance of lung adenocarcinoma PC9/GR cells and its mechanism].

Objective: To screen the key genes involved in gefitinib resistance of lung adenocarcinoma PC9/GR cells which harbored 19 exon mutation of epidermal growth factor receptor (EGFR) gene, and discuss the effect and mechanism of downregulation of solute carrier family 7 member 11 (SLC7A11) on the gefitinib resistance of PC9/GR cells. Methods: RNA microarray was conducted to detect the gene expressions in PC9 and PC9/GR cells. The differently expressed genes were screened by using limma package of R language and analyzed by Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Western blotting was performed to determine the expression of SLC7A11 protein in PC9 and PC9/GR cells. PC9/GR cells were infected with lentivirus plasmid containing short hairpin RNA (shRNA) targeting SLC7A11 or negative control shRNA (sh-NC), respectively. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the efficacy of shRNA on the expression of SLC7A11 mRNA. Cell counting kit-8 (CCK-8) assay was conducted to determine the suppressing effect of gefitinib on PC9/GR cells. Mito-Tracker Red CMXRos probe and malondialdehyde (MDA) assay kit were used to evaluate gefitinib-induced ferroptosis in PC9/GR cells. Immunohistochemistry (IHC) was conducted to detect the expression of SLC7A11 protein in the tumor tissues of advanced stage lung adenocarcinoma patients harboring 19 exon mutation of EGFR gene. Thirty-six advanced stage lung adenocarcinoma patients who received EGFR-tyrosihe kinase inhibitor(TKI) as first-line treatment in Fourth Hospital of Hebei Medical Unviersity were enrolled. Kaplan-Meier survival curve was drawn to analyze the correlation between SLC7A11 expression and progression-free survival (PFS) of the patients. Results: RNA array demonstrated that 2 888 genes were differently expressed between PC9 and PC9/GR cells. KEGG analysis showed that ferroptosis-related gene was one of the most enriched region of the differently expressed genes between PC9 and PC9/GR cells. These ferroptosis-related gene cohort contained 13 genes, among which SLC7A11 exhibited the most significant difference. Western blotting showed that the expression of SLC7A11 protein in PC9/GR cells was significantly higher than that in PC9 cells (0.76±0.03 vs. 0.19±0.02, P<0.001). The 50% inhibiting concentration (IC(50)) of gefitinib was 35.08 µmol/L and 64.01 µmol/L for sh-SLC7A11 and sh-NC group PC9/GR cells, respectively. PC9/GR cells in sh-SLC7A11 group exhibited significantly lower density of mitochondria fluorescence after gefitinib treatment, compared to the sh-NC group (213.77±26.50 vs. 47.88±4.55, P<0.001). In addition, PC9/GR cells in sh-SLC7A11 group exhibited significantly higher MDA after gefitinib treatment, compared to the sh-NC group [(15.43±1.60) µmol/mg vs. (82.18±7.77) µmol/mg, P<0.001]. The PFS of the patients with low expression of SLC7A11 (n=18) was significantly longer than the patients with high expression of SLC7A11 (n=18, 16.77 months vs. 9.14 months, P<0.001). Conclusion: Downregulation of SLC7A11 could increase the sensitivity of PC9/GR cells to gefitinib by promoting ferroptosis.

Ru-Based Organometallic Agents Bearing Phenyl Hydroxide: Synthesis and Antibacterial Mechanism Study against Staphylococcus aureus.

The development of antimicrobial agents with novel model of actions is a promising strategy to combat multiple resistant bacteria. Here, three ruthenium-based complexes, which acted as potential antimicrobial agents, were synthesized and characterized. Importantly, three complexes all showed strong bactericidal potency against Staphylococcus aureus. In particular, the most active one has a MIC of 6.25 µg/mL. Mechanistic studies indicated that ruthenium complex killed S. aureus by releasing ROS and damaging the integrity of bacterial cell membrane. In addition, the most active complex not only could inhibit the biofilm formation and hemolytic toxin secretion of S. aureus, but also serve as a potential antimicrobial adjuvant as well, which showed synergistic effects with eight traditional antibiotics. Finally, both G. mellonella larva infection model and mouse skin infection model all demonstrated that ruthenium complex also showed significant efficacy against S. aureus in vivo. In summary, our study suggested that ruthenium-based complexes bearing a phenyl hydroxide are promising antimicrobial agents for combating S. aureus.

[Current status of diagnosis and treatment of chronic lymphocytic leukemia in China: A national multicenter survey research].

Objective: To understand the current status of diagnosis and treatment of chronic lymphocytic leukemia (CLL) /small lymphocytic lymphoma (SLL) among hematologists, oncologists, and lymphoma physicians from hospitals of different levels in China. Methods: This multicenter questionnaire survey was conducted from March 2021 to July 2021 and included 1,000 eligible physicians. A combination of face-to-face interviews and online questionnaire surveys was used. A standardized questionnaire regarding the composition of patients treated for CLL/SLL, disease diagnosis and prognosis evaluation, concomitant diseases, organ function evaluation, treatment selection, and Bruton tyrosine kinase (BTK) inhibitor was used. Results: ①The interviewed physicians stated that the proportion of male patients treated for CLL/SLL is higher than that of females, and the age is mainly concentrated in 61-70 years old. ②Most of the interviewed physicians conducted tests, such as bone marrow biopsies and immunohistochemistry, for patient diagnosis, in addition to the blood test. ③Only 13.7% of the interviewed physicians fully grasped the initial treatment indications recommended by the existing guidelines. ④In terms of cognition of high-risk prognostic factors, physicians' knowledge of unmutated immunoglobulin heavy-chain variable and 11q- is far inferior to that of TP53 mutation and complex karyotype, which are two high-risk prognostic factors, and only 17.1% of the interviewed physicians fully mastered CLL International Prognostic Index scoring system. ⑤Among the first-line treatment strategy, BTK inhibitors are used for different types of patients, and physicians have formed a certain understanding that BTK inhibitors should be preferentially used in patients with high-risk factors and elderly patients, but the actual use of BTK inhibitors in different types of patients is not high (31.6%-46.0%). ⑥BTK inhibitors at a reduced dose in actual clinical treatment were used by 69.0% of the physicians, and 66.8% of the physicians had interrupted the BTK inhibitor for >12 days in actual clinical treatment. The use of BTK inhibitors is reduced or interrupted mainly because of adverse reactions, such as atrial fibrillation, severe bone marrow suppression, hemorrhage, and pulmonary infection, as well as patients' payment capacity and effective disease progression control. ⑦Some differences were found in the perceptions and behaviors of hematologists and oncologists regarding the prognostic assessment of CLL/SLL, the choice of treatment options, the clinical use of BTK inhibitors, etc. Conclusion: At present, a gap remains between the diagnosis and treatment of CLL/SLL among Chinese physicians compared with the recommendations in the guidelines regarding the diagnostic criteria, treatment indications, prognosis assessment, accompanying disease assessment, treatment strategy selection, and rational BTK inhibitor use, especially the proportion of dose reduction or BTK inhibitor discontinuation due to high adverse events.

[A case report of chronic granulomatous disease with CYBB gene mutation in an adult].

We reported a 28-year-old male patient who had been admitted to a local hospital for several times in the past four years because of recurrent fever and cough. Each chest CT scan during hospitalization showed consolidation accompanied by exudation and mild pleural effusion. After treatment, the consolidation apparently absorbed, but similar symptoms recurred within half a year, and the new consolidation appeared. For this reason, he was diagnosed with tuberculosis or bacterial pneumonia several times in other hospitals, and was hospitalized two to three times a year. Finally, he was diagnosed with chronic granulomatous disease (CGD) with CYBB gene mutation through whole-exome sequencing.

Cefoperazone/sulbactam-induced hemolytic anemia.

Drug-induced hemolytic anemia (DIHA) is a rare complication of drug therapy and usually underdiagnosed. Cefoperazone/sulbactam is a compound prepared from the third generation of cephalosporin and ß-lactamase inhibitor. There are limited data of DIHA induced from cefoperazone/sulbactam. A 93-year-old female patient, who had an operation on the biliary tract 3 months ago, was admitted to our hospital with an abdominal infection. After cefoperazone/sulbactam was given as anti-infection treatment, the patient developed hemolytic anemia on the third day. Cefoperazone/sulbactam was discontinued and replaced with meropenem. Subsequently the level of red blood cells, hemoglobin, and hematocrit returned to normal. Clinicians should pay attention to monitoring the possible adverse reactions during the use of cefoperazone/sulbactam and should be aware of the occurrence of DIHA, so as to give timely treatment.

[Progress on clinical trials of common gastrointestinal cancer drugs in China from 2012 to 2021].

Objective: Systematically summarize the research progress of clinical trials of gastric cancer oncology drugs and the overview of marketed drugs in China from 2012 to 2021, providing data and decision-making evidence for relevant departments. Methods: Based on the registration database of the drug clinical trial registration and information disclosure platform of Food and Drug Administration of China and the data query system of domestic and imported drugs, the information on gastric cancer drug clinical trials, investigational drugs and marketed drugs from January 1, 2012 to December 31, 2021 was analyzed, and the differences between Chinese and foreign enterprises in terms of trial scope, trial phase, treatment lines and drug type, effect and mechanism studies were compared. Results: A total of 114 drug clinical trials related to gastric tumor were registered in China from 2012 to 2021, accounting for 3.7% (114/3 041) of all anticancer drug clinical trials in the same period, the registration number showed a significant growth rate after 2016 and reached its peak with 32 trials in 2020. Among them, 85 (74.6%, 85/114) trials were initiated by Chinese pharmaceutical enterprise. Compared with foreign pharmaceutical enterprise, Chinese pharmaceutical enterprise had higher rates of phase I trials (35.3% vs 6.9%, P=0.001), but the rate of international multicenter trials (11.9% vs 67.9%, P<0.001) was relatively low. There were 76 different drugs involved in relevant clinical trials, of which 65 (85.5%) were targeted drugs. For targeted drugs, HER2 is the most common one (14 types), followed by PD-1 and multi-target VEGER. In the past ten years, 3 of 4 marketed drugs for gastric cancer treatment were domestic and included in the national medical insurance directory. Conclusions: From 2012 to 2021, China has made some progress in drug research and development for gastric carcinoma. However, compared with the serious disease burden, it is still insufficient. Targeted strengthening of research and development of investment in many aspects of gastric cancer drugs, such as new target discovery, matured target excavating, combination drug development and early line therapy promotion, is the key work in the future, especially for domestic companies.

Protective effect of miR-146 against kidney injury in diabetic nephropathy rats through mediating the NF-κB signaling pathway.

The article "Protective effect of miR-146 against kidney injury in diabetic nephropathy rats through mediating the NF-κB signaling pathway", H.-Y. Yu, L.-F. Meng, X.-H. Lu, L.-H. Liu, X. Ci, Z. Zhuo, published in Eur Rev Med Pharmacol Sci 2020; 24 (6): 3215-3222-DOI: 10.26355/eurrev_202003_20688-PMID: 32271439, has been retracted by the authors due to some inaccuracies (some data cannot be repeated by further research). The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20688.

[Clinical effect analysis of 146 cases of ipsilateral simultaneous pancreas and kidney transplantation].

Objective: To investigate the clinical effect of ipsilateral simultaneous pancreas and kidney transplantation (SPK). Methods: A total of 146 cases of SPK surgeries completed in the Second Affiliated Hospital of Guangzhou Medical University from September 2016 to June 2020 were selected to summarize the outcome, curative effect and complications of the operation. Results: The patients were followed up for 1 to 45 months. Good clinical results were obtained in 146 patients. Renal function indicators suggest that on the 7th day after operation, the serum creatinine returned to normal level [142.4 (108.6, 213.4)µmol/L]. The index of pancreatic function decreased to the normal level as expected. The level of blood amylase was 160.5(109.3, 249.8) U/L within 7 days after operation, and then decreased. The trend of urinary amylase was similar to that of blood amylase, which was 240(121.0, 370.0) U/L 7 days after operation, and glycosylated hemoglobin decreased to the normal level (5.8%±1.4%) 1 month after operation. The main medical complications were infection including pulmonary infection (26.03%, 38/146), urinary tract infection (26.03%,38/146), and abdominal infection (4.79%,7/146), acute rejection including renal graft rejection (5.8%,8/146), pancreas/duodenum rejection (18.49%,27/146), and renal graft combined pancreatic graft rejeciton (6.85%,10/146), as well as gastrointestinal bleeding (30.82%,45/146), of which 5 cases were severe bleeding (3.42%, 5/146). The main surgical complications were poor incision healing (10.27%, 15/146), serious surgical complications including arteriovenous thrombosis of the transplanted pancreas (2.05%, 3/146) and intestinal leakage (0.68%,1/146). The 1-year and 3-year patient, renal and pancreatic survival rates were both 92.5%, 91.5% and 89.5%, respectively, and despite the death, the 1-year, 3-year transplanted kidney survival rate was both 99.3%, and 95% for the the 1-year, 3-year pancreas survival rate. Conclusion: Strict preoperative evaluation of the function of large organs, reasonable surgical methods, perioperative anticoagulation, and prompt diagnosis of complications can achieve good clinical results for patients with SPK.

[Application of metabolomics in nanotoxicity].

As an emerging material with excellent property, nanoparticle has been widely used in multiple fields such as chemical industry, textile, biomedicine, etc., which has caused widespread concern about its potential toxicity in society. Due to the limitations of traditional toxicology technology, the high-throughput methods were urgently needed to assess the potential toxicity of particles at nanoparticle size systematically. Metabolomics has been recognized as an emerging omics technology developed by multidisciplinary combination of modern analytical techniques, biochemistry and bioinformatics, which has been widely used to screened early toxicity potential markers and metabolic pathway by analyzing changes in the types and quantities of metabolites in biological samples such as cells and tissues. This article reviewed the progress and challenges of metabolomics in nanotoxicology research.

[Current status and prospects of population exposure assessment of nanomaterials consumer products].

With the wide application of nanomaterials in consumer products in the market, it is necessary to understand the existence and release of nanomaterials in consumer products, as well as the current situation of exposure assessment of consumers. China has been a large industrial producer with a huge consumer market, but the supervision of consumer goods with nanomaterials is almost blank. This article summarized and classified the existing consumer products of nanomaterials in the international market, and discussed the release of key nanomaterials in consumer products and the exposure assessment methods of consumers, in order to provide a scientific basis for the establishment of a regulatory system for consumer products of nanomaterials in China in the future.

[Application effects of self-made simple vacuum sealing drainage device in postoperative treatment of sural neurocutaneous flap transplantation in the foot and ankle].

Objective: To investigate the application effects of self-made simple vacuum sealing drainage (VSD) device in the postoperative treatment of sural neurocutaneous flap transplantation in the foot and ankle. Methods: From January 2017 to January 2019, 36 patients with foot and ankle skin defects and bone exposure admitted to People's Hospital of Xinjiang Uygur Autonomous Region met the inclusion criteria, and a retrospective cohort study was conducted. According to the bandaging method of the operative area, simple negative pressure group and antibacterial dressing group were both allocated with 18 patients, with 12 males and 6 females in the former group, 14 males and 4 females in the latter group, aged (41.6±2.8) and (42.3±2.6) years, respectively. Patients in the two groups all received sural neurocutaneous flap transplantation. Patients in antibacterial dressing group received nano silver antibacterial dressing change in the operative area, and the dressing was changed once every 3 days. In simple negative pressure group, the operative area was sealed with a simple VSD device made of gauze, silicone sputum suction tube with holes cut out, and biological permeable membrane, etc., which was connected with the wall central negative pressure suction system for continuous VSD treatment of -40.0 to -16.6 kPa. The negative pressure material was changed once every 5 days. The number of dressing change, the pain score evaluated by Numeric Rating Scale during each dressing change, the cost of dressing change, and the degree of flap swelling evaluated on the 3rd and 5th day after surgery were recorded, and the flap survival was observed. Data were statistically analyzed with independent sample t test, Wilcoxon rank sum test, and chi-square test. Results: The number of dressing change of patients in simple negative pressure group was (3.4±0.5) times, which was significantly less than (7.0±0.8) times in antibacterial dressing group (t=15.338, P<0.01). The pain score during dressing change of patients in simple negative pressure group was (4.3±0.8) points, which was significantly lower than (6.8±0.7) points in antibacterial dressing group (t=10.168, P<0.01). The cost of dressing change of patients was similar between the two groups. On the 3rd and 5th day after surgery, the degrees of flap swelling of patients in simple negative pressure group were significantly superior to those in antibacterial dressing group (Z=4.448, 2.395, P<0.05 or P<0.01). The flap survival of patients in simple negative pressure group was significantly superior to that in antibacterial dressing group (χ(2)=4.500, P<0.05). Conclusions: Compared with the traditional dressing bandage, the self-made simple VSD device used after sural neurocutaneous flap transplantation can reduce the frequency of dressing change, relieve the pain of dressing change and the swelling of flap, and promote the flap survival, which is worth popularizing and applying in clinic.

Study on the role and mechanism of ß4GalT1 both in vivo and in vitro glioma.

OBJECTIVE: To discuss the role and mechanism of ß4GalT1 both in vivo and in vitro glioma, observe whether pathophysiological processes of glioma can be improved after ß4GalT1 is knocked down, and study whether ß4GalT1 plays a role in malignant biological processes of glioma by regulating the apoptosis and immune processes. PATIENTS AND METHODS: Firstly, the distribution difference of ß4GalT1 in tumor tissues and normal tissues was analyzed by Gene Expression Profiling Interactive Analysis (GEPIA) tumor analysis system to deduce the possible role of ß4GalT1 in glioma. Secondly, whether the malignant degree of glioma was related to the expression of ß4GalT1 and its immunity using human tumor tissues and blood lymphocyte subsets was analyzed. Thirdly, interfere lentivirus vector with ß4GalT1 and knockdown ß4GalT1 was analyzed to observe whether the malignant degree of glioma has changed. Fourthly, interfere lentivirus vector with recombinant ß4GalT protein and ß4GalT1 was analyzed to verify the effect of ß4GalT in vitro test. Fifth, interfere lentivirus vector with recombinant ß4GalT protein and ß4GalT1 was analyzed to verify effect of ß4GalT in vivo test. Finally, we discuss whether ß4GalT is involved in the biological process of glioma through inflammatory reaction. RESULTS: In the GEPIA tumor analysis system, the expression in tumor was significantly higher than that in normal tissues. The expression of ß4GalT1 in glioma tissues was higher than that in normal tissues, and the higher the malignancy of the tumor, the higher the expression of ß4GalT1 in the glioma tissues, and the lower the immune level was. The expression of IDH1, MGMT, and ki-67 was reduced, and the survival rate of the mice with glioma was improved after ß4GalT1 was knocked down. In vitro tests, the activity of tumor cells and their reproductive ability can be reduced after ß4GalT1 was knocked down, the immune level of the body can be improved, and the level of tissue apoptosis can be reduced. After recombinant ß4GalT1 was given alone, the result was opposite to that of ß4GalT1 knocked down group. In vivo tests, gross tumor volume can be reduced after ß4GalT1 was knocked down, the immune level of the body can be improved, and the level of tissue apoptosis can be reduced. After recombinant ß4GalT1 was given alone, the result was opposite to that of ß4GalT1 knocked down group. After knocking down ß4GalT1, the expression of inflammatory factors can be reduced both in vivo and in vitro, and the inflammatory microenvironment of tumors can be improved. After recombinant ß4GalT1 was given alone, the result was opposite to that of ß4GalT1 knocked down group. CONCLUSIONS: The level of ß4GalT1 expression in tumor tissues was increased. The malignant degree of glioma is related to the expression of ß4GalT1 and its immunity. The level of tumor marker can be decreased, and the survival rate of glioma model mice can be increased after ß4GalT1 is knocked down. Apoptosis and immune injury caused by tumor can be improved and gross tumor volume can be deduced after ß4GalT1 is knocked down. During the development of glioma, ß4GalT1 may play a malignant biological role through inflammatory response.

[A pair of baby twins with severe burns simultaneously].

December 22, 2018, a pair of female baby twins aged 2 years with severe burns at the same time were transferred from the primary hospital to the People's Hospital of Xinjiang Uygur Autonomous Region on day 17 post burn. Physical examination on admission showed that both of them had severe burns on head, face, left upper limb, foot, etc. The blood preparations were difficult for them due to their rare blood types, with the elder sister of type B blood combined with Rh D mutant and the younger sister of type B combined with Rh negative blood. The total area of burn of the younger sister was 15% total body surface area (TBSA), with full-thickness burns with tendon and bone exposure in left ear and most of the ear auricle fallen off. The younger sister was treated with anti-infection, anti-shock by fluid infusion, and nutritional support. Split-thickness skin graft on left thigh was harvested to repair wound on left upper limb, and thin medium-thickness skin graft was harvested from right thigh in different operations to repair wounds on head and face, and thin medium-thickness skin graft from right thigh with area of 3 cm×1 cm was reserved. The total area of burn of the elder sister was 22% TBSA, combined with inhalation injury and severe lung infection after tracheotomy, and the condition was critical. After admission to the hospital, anti-infection, anti-shock by fluid infusion, nutritional support, atomization, and eliminating phlegm treatment were performed. After the general condition was basically stable and blood preparation was sufficient, thin medium-thickness skin grafts were harvested from left thigh to repair wounds on head, face, and left upper limb, and the reserved thin medium-thickness skin graft from right thigh of the younger sister was transferred to the posterior ear wound of the elder sister. The skin grafts survived after operation with expansion showing no difference from autogenous skin. The elder sister and the younger sister were discharged from the hospital on the 24 and 21 day of hospitalization respectively. During follow-up of 6 months, the effect of the operation was good.

Protective effect of miR-146 against kidney injury in diabetic nephropathy rats through mediating the NF-κB signaling pathway.

OBJECTIVE: To study the protective effect of micro ribonucleic acid (miR)-146 against kidney injury in diabetic nephropathy (DN) rats through the nuclear factor-κB (NF-κB) signaling pathway. MATERIALS AND METHODS: In this experiment, 30 adult Sprague-Dawley rats with 5-6 weeks old and weighing 20-30 g were selected and randomly divided into control group (n=10), model group (n=10), and miR-146 Mimic group (n=10, DN rat model + miR-146 Mimic). The serum levels of creatinine (Cr) and blood urea nitrogen (BUN) in the three groups were detected using the full-automatic biochemical analyzer. The protein expression levels of phosphorylated-inhibitor of NF-κB (p-IκB), p-P65, P65, and Tubulin were detected via Western blotting. The messenger RNA (mRNA) of P65 was determined using quantitative Polymerase Chain Reaction (qPCR). Positive expression of p-IκB in tissues was determined using immunohistochemistry. Moreover, the contents of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and IL-6 were detected using the enzyme-linked immunosorbent assay (ELISA) kits. Finally, the apoptosis was detected through Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) dual-fluorescence labeling. RESULTS: The serum levels of Cr and BUN were significantly higher in the model group than those in the control group (p<0.01), while they were significantly lower in miR-146 Mimic group than those in the model group (p<0.05). The levels of p-IκB and p-P65/P65 significantly increased in the model group compared with those in the control group (p<0.01), while they remarkably declined in the miR-146 Mimic group compared with those in the model group (p<0.05). The results of qPCR showed that the mRNA level of P65 had no significant difference among the three groups (p>0.05). The immunohistochemical assay showed that the positive expression of p-IκB in tissues was consistent with those of the protein level as Western blotting revealed. The rats in the model group had evidently increased levels of TNF-α, IL-1ß, and IL-6 compared with the control group (p<0.01), while miR-146 Mimic group had evidently decreased levels of them compared with the model group (p<0.01). Finally, apoptosis was enhanced in the model group compared with that in the control group, while it was remarkably inhibited in the miR-146 Mimic group. CONCLUSIONS: MiR-146 can inhibit the NF-κB signaling pathway, lower the levels of TNF-α, IL-1ß, and IL-6, and reduce the apoptosis, thereby exerting a protective effect against kidney injury in DN.

Effect of lncRNA AK125437 on postmenopausal osteoporosis rats via MAPK pathway.

OBJECTIVE: The aim of this study was to explore the effect of long non-coding ribonucleic acid (lncRNA) AK125437 on rats with postmenopausal osteoporosis via the mitogen-activated protein kinase (MAPK) pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly divided into three groups, including normal group, model group, and an inhibitor group, with 12 rats in each group. Only ovaries were exposed in normal group. The postmenopausal osteoporosis model was established in model group. Meanwhile, the intervention was performed with inhibitor for 3 months after modeling in inhibitor group, followed by sampling. The expression of receptor activator of nuclear factor kappa-B ligand (RANKL) was detected via immunohistochemistry. The protein expression level of phosphorylated p38 (p-p38) MAPK was determined via Western blotting (WB). Furthermore, the expression level of lncRNA AK125437 and the content of serum estradiol were determined via quantitative Polymerase Chain Reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively. In addition, bone mineral density was measured using dual-energy X-ray bone mineral absorptiometer. RESULTS: Immunohistochemistry results indicated that model group and inhibitor group had notably up-regulated positive expression level of RANKL than normal group (p<0.05), which was remarkably lower in inhibitor group than model group (p<0.05). Western blot results showed that compared with normal group, the protein expression level of p-p38 MAPK was substantially elevated in model and inhibitor groups (p<0.05). Meanwhile, the protein expression level of p-p38 MAPK was markedly lower in inhibitor group than that in model group (p<0.05). According to qPCR results, the expression level of lncRNA AK125437 was significantly up-regulated in both model group and inhibitor group compared with normal group, showing statistically significant differences (p<0.05). However, no significant differences were observed between model group and inhibitor group (p>0.05). ELISA results revealed that model group and inhibitor group had markedly lower estradiol content than normal group (p<0.05). There was no statistically significant difference in the content of estradiol between the two groups (p>0.05). According to the measurement results of bone mineral density, compared with normal group, bone mineral density was notably lower in model group and inhibitor group (p<0.05). Furthermore, it was markedly higher in inhibitor group than that of model group (p<0.05). CONCLUSIONS: LncRNA AK125437 affects the bone mineral density of rats with postmenopausal osteoporosis by activating the MAPK pathway.

Melatonin reduces OGD/R-induced neuron injury by regulating redox/inflammation/apoptosis signaling.

OBJECTIVE: To investigate the roles and underlying mechanisms of melatonin in oxygen-glucose deprivation/reoxygenation (OGD/R)-insulted SH SY5Y cells. MATERIALS AND METHODS: SH SY5Y cells were cultured for OGD/R stimulation. Cell viability and cytotoxicity were measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, lactate dehydrogenase (LDH), and Hoechst 33258/propidium iodide (PI) staining assays. The mRNA levels of high mobility group box-1 (HMGB1), tumor necrosis factor α (TNF-α), and inducible nitric oxide synthase (iNOS) were analyzed by quantitative Real Time-PCR assays. Nitric oxide (NO) production was assessed by Griess reagent. Reactive oxygen species (ROS) production was detected by fluorescent probe. Malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were examined by commercial kits. Cell apoptosis was analyzed by flow cytometry and caspase-3 activity. The protein levels were detected by Western blot. RESULTS: Melatonin enhanced the viability and reduced the death and LDH release of OGD/R exposed SH SY5Y cells. Melatonin repressed the HMGB1, TNF-α, and iNOS mRNA expression, NO production, and nuclear factor κB (NF-κB) activation in OGD/R challenged SH SY5Y cells. Melatonin reduced the ROS, MDA, 4-HNE, and 8-OHdG contents but further enhanced the levels of the nuclear factor E2-related factor-2 (Nrf2) and heme oxygenase (HO-1). Melatonin-increased viability and melatonin-decreased LDH release were also mediated by the blockage of NF-κB or reversed by Nrf2 or HO-1 knockdown. Melatonin exerted antiapoptotic effect on OGD/R treated SH SY5Y cells partly by activating Akt signaling. OGD/R challenged SH SY5Y cell autophagy was also repressed by melatonin, as evidenced by the decreased levels of LC-II and beclin-1 and the increased phosphorylation of mammalian target of rapamycin (mTOR), p70 ribosomal protein S6 kinase (p70S6K), and eukaryotic initiation factor 4E binding protein 1 (4E-BP-1). CONCLUSIONS: Melatonin protected SH SY5Y cells from OGD/R induced oxidative stress, inflammation, apoptosis, and autophagy by blocking NF-κB signaling and activating Nrf2/HO-1, Akt, and mTOR/p70S6K/4E-BP-1 pathways, thereby indicating that melatonin is a potential and novel therapeutic drug for ischemic stroke.

Implementation of a classification strategy of Raman data collected in different clinical conditions: application to the diagnosis of chronic lymphocytic leukemia.

The literature is rich in proof of concept studies demonstrating the potential of Raman spectroscopy for disease diagnosis. However, few studies are conducted in a clinical context to demonstrate its applicability in current clinical practice and workflow. Indeed, this translational research remains far from the patient's bedside for several reasons. First, samples are often cultured cell lines. Second, they are prepared on non-standard substrates for clinical routine. Third, a unique supervised classification model is usually constructed using inadequate cross-validation strategy. Finally, the implemented models maximize classification accuracy without taking into account the clinician's needs. In this paper, we address these issues through a diagnosis problem in real clinical conditions, i.e., the diagnosis of chronic lymphocytic leukemia from fresh unstained blood smears spread on glass slides. From Raman data acquired in different experimental conditions, a repeated double cross-validation strategy was combined with different cross-validation approaches, a consensus label strategy and adaptive thresholds able to adapt to the clinician's needs. Combined with validation at the patient level, classification results were improved compared to traditional strategies.