Objective To investigate the expression and subcellular localization of long non-coding RNA (lncRNA)COL11A1-208, as well as the effect on the proliferation and invasion of oral squamous cell carcinoma (OSCC) cells. Methods The differential expression of COL11A1-208 in seven OSCC cell lines compared with primary cultured normal oral epithelial cells was detected by quantitative real-time PCR (qPCR), and the subcellular localization of COL11A1-208 was evaluated by cell nuclear/cytoplasmic fractionation and fluorescence in situ hybridization (FISH). CAL27 and HN4 cells were respectively transfected with COL11A1-208 Smart Silencer (SS-COL11A1-208 group) and negative control (NC) Smart Silencer (SS-NC group), while HN4 and HN6 cells were stably infected by COL11A1-208 lentivirus overexpression vector (LV-COL11A1-208 group) and NC lentivirus overexpression vector (LV-NC group). Cell proliferation in each group was detected by CCK-8 assay and plate colony formation assay, while cell migration and invasion ability in each group were detected by Transwell assays. Then, the expression of COL11A1 were detected by qPCR and Western blotting assays, while the protein expression of E-cadherin and vimentin was detected by Western blotting assay. Finally, the effect of COL11A1-208 on OSCC cells in vivo was studied by xenograft formation assay. Results COL11A1-208 was highly expressed in OSCC cell lines compared with the normal cells (P<0.05). The results of nuclear and cytoplasmic RNA isolation assay showed that the nuclear proportion of COL11A1-208 in CAL27, HN4 and HN6 cells was significantly higher than that in cytoplasm (P<0.01); similarly, FISH results showed that COL11A1-208 was primarily localized within the nucleus of OSCC cells. The relative expression of COL11A1-208 in SS-COL11A1-208 group was significantly lower than that in the SS-NC group (CAL27: 0.225±0.030 vs. 1.000±0.000; HN4: 0.393±0.028 vs. 1.000±0.000; P<0.01). Compared with the SS-NC group, proliferation activity, colonizing ability, migration and invasion abilities of SS-COL11A1-208 group decreased significantly (P<0.05). The relative expression of COL11A1-208 in LV-COL11A1-208 group was significantly higher than that in the LV-NC group (HN6: 6524.216±3395.926 vs. 1.000±0.000; HN4: 3486.230±743.908 vs. 1.000±0.000; P<0.05).Compared with the LV-NC group, proliferation activity, colonizing ability, migration and invasion abilities of LV-COL11A1-208 group were significantly increased (P<0.05). Compared with the SS-NC group, the relative expression of COL11A1 protein in SS-COL11A1-208 group decreased significantly (P<0.05), while the relative expression of COL11A1 protein in LV-COL11A1-208 group increased significantly than that in the LV-NC group (P<0.01). In addition, compared with the SS-NC group, the relative expression of E-cadherin in SS-COL11A1-208 group increased significantly (P<0.05), and the relative expression of vimentin decreased significantly (P<0.05). In LV-COL11A1-208 group, the relative expression of E-cadherin decreased significantly (P<0.05),and the relative expression of vimentin increased significantly (P<0.01). In vivo experiments showed that the xenograft tumor weight and volume of ASO-COL11A1-208 group were significantly reduced (P<0.05), while the xenograft weight and volume of LV-COL11A1-208 group were significantly enhanced (P<0.05). Conclusion COL11A1-208 could facilitate the cell proliferation and invasion of OSCC, which plays an important role in the development and progression of OSCCs.

Objective To investigate the role and significance of abdominal paracentesis drainage (APD) to pyroptosis in the pancreas of rats with severe acute pancreatitis (SAP), which is mediated by NLRP3 inflammasome-activated-caspase-1. Methods A total of 48 healthy male SD rats were randomly divided into three groups: sham operation (Sham) group, SAP group, and APD group, with 16 rats in each group. 5% sodium sulfonate was retrogradely injected into the pancreaticobiliary duct to establish the SAP model. In the APD group, besides the SAP induction, a drainage tube was placed on the right lower abdomen for drainage. Serum and pancreatic tissue were collected 12 hours after modeling. We examined the activities of serum lipase and amylase using an automatic biochemical analyzer and evaluated pancreatic damage through HE staining. In addition, we quantified the expression of serum inflammatory factors by ELISA and measured the expression level of genes and proteins related to caspase-1 mediated pyroptosis in pancreatic tissue using RT-PCR and Western blotting. Lastly, we observed the structural changes of the subcellular organelles and the characteristic changes of pyrolysis in pancreatic acinar cells using transmission electron microscopy. Results Compared with the SAP group, the pancreas tissue in the ADP group showed alleviated damages with a much lower pathological score; the levels of serum lipase, amylase, tumor necrosis factor(TNF)-α, Interleukin(IL)-6, IL-1β, and IL-18 also showed significant reduction (P<0.05).Compared with the Sham group, the expression levels of NLRP3, ASC, Caspase-1, cleaved caspase-1, GSDMD, and cleaved-GSDMD were all significantly up-regulated in the pancreas in the SAP group. After APD treatment, the expression of these Caspase-1-mediated pyrolysis pathway key genes in pancreatic tissue was significantly down-regulated. In addition, the expression levels of IL-1β and IL-18 mRNA in pancreatic tissue were also significantly reduced (P<0.05). Transmission electron microscopy showed that the SAP group had endoplasmic reticulum and mitochondrial expansion, chromatin condensation, and the characteristic change of pyrolysis, namely the formation of membrane pores in the cell membrane. In the APD group, we observed reduced chromatin condensation in the nucleus and less expansion in the endoplasmic reticulum and mitochondrial. Conclusion SAP showed activated Caspase-1-mediated pyroptosis. Early treatment with APD can alleviate the severity of SAP, possibly by inhibiting the activation of Caspase-1-mediated pyroptosis, thereby reducing local and systemic inflammatory reactions.

Objective To study the effect of Fritillaria cirrhosae bulbus on ovalbumin (OVA)-sensitized asthmatic mice and its mechanism. Methods We set up the study including a blank control group, OVA model group (intraperitoneal injection of 0.2 mg OVA to induce sensitization), dexamethasone group (after successful modeling, intraperitoneal injection of 0.5 mg/kg dexamethasone), and Fritillaria cirrhosae bulbus low, medium and high dose groups (after successful modeling, gavage 7.0 mg/kg,14.0 mg/kg, 21.0 mg/kg Fritillaria cirrhosae bulbus), once a day for 4 weeks, with 10 mice in each group (total=60 BALB/c mice).After drug administration, we measured the amount of phenol red excretion in the airway and collected the bronchoalveolar lavage fluid (BALF) to determine the content of brain-derived neurotrophic factor (BDNF) and the number of inflammatory cells. We also took the blood samples through eyeballs followed by quantifying the concentration of serum immunoglobulin E (IgE), interleukin-4(IL-4), IL-13, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) using ELISA. We further collected the lung tissues and detected the mRNA and protein expressions of Janus kinase 3 (JAK3), signal transducer and activator of transcription 6 (STAT6),and IL-4 by RT-qPCR and Western blotting. Results Compared with the blank control group, the level of phenol red in the airway of the OVA model group reduced significantly, the content of BDNF as well as the number of inflammatory cells in BALF increased significantly, the contents of IgE, IL-4, IL-13, and TNF-α in serum were significantly elevated, the concentration of IFN-γ and IFN-γ/IL-4 ratio decreased significantly, the relative expression levels of JAK3, STAT6, IL-4 mRNA and p-JAK3/JAK3, p-STAT6/STAT6, and IL-4 protein in lung tissue increased significantly (P<0.05). Compared with the OVA model group, the above indexes were all significantly improved in each dose of the Fritillaria cirrhosae bulbus groups, and the high Fritillaria cirrhosae bulbus dose group had the best index (P<0.05). Conclusion The mechanism of Fritillaria cirrhosae bulbus regulating airway and lung inflammation in asthmatic mice may be related to inhibiting the activation of JAK3/STAT6 signal pathway.

Objective To investigate the effect and mechanism of verteporfin (Vp) in airway remodeling of experimental asthma. Methods (1) Animal experiment: Twelve female BALB/c mice were divided into 3 groups (4 each): control group experimental asthma group and Vp intervention asthma group. Mice in experimental asthma group were treated by intranasal delivery of house dust mite (HDM) extract, in Vp intervention group were treated with intraperitoneal injection of Vp before HDM stimulation, and in control group was treated with the same amount of normal saline. Lung tissues and bronchoalveolar lavage fluid (BALF) of mice were collected, HE staining, Masson staining and Periodic Acid-Schiff (PAS) staining were used to observe the pathological changes of lung tissue, Wright-Giemsa staining was used to count the inflammatory cells in BALF, and the expressions of Yes-associated protein (YAP) and phosphorylated Yes-associated protein (p-YAP) in lung tissues were detected with immunohistochemistry, RT-PCR and Western blotting were performed to detect the expressions of YAP, p-YAP, osteopontin (OPN)and smooth muscle myosin heavy chain (SMMHC). (2) Cell experiment: Human airway smooth muscle cells (HASMC) were divided into control group (treated with the same amount of normal saline), HDM group (treated with 50 μg/ml HDM for 24 hours)and HDM+Vp group (treated with 0.05 mg/L Vp for 2 hours, and then with 50 μg/ml HDM for 24 hours). The cell proliferation of HASMC was detected by CCK-8. The apoptosis rate of HASMC were detected by flow cytometry. The protein levels of YAP, OPN and SMMHC in cells were detected by Western blotting. Results (1) Animal experiment: Compared with control group, the lung inflammation was aggravated, the bronchial basement membrane, airway smooth muscle (ASM) and airway wall were thickened in experimental asthma group [(1.13±0.38) μm vs. (0.79±0.36) μm, (6.49±2.36) μm vs. (4.56±1.52) μm, (33.85±5.95) μm vs. (22.08±3.30) μm, P<0.05]. The deposition proportion of collagen fiber around airway and PAS staining area increased(5.85%±2.35% vs. 0.36%±0.12%, 28.81%±5.89% vs. 13.57%±2.08%, P<0.01). The mRNA and protein expression levels of YAP and OPN increased (P<0.05 or P<0.01), and the ratio of p-YAP/YAP and the mRNA and protein expression levels of SMMHC decreased (P<0.05 or P<0.01). Compared with experimental asthma group, the lung inflammation was reduced, and the thickness of bronchial basement membrane, ASM and airway wall were thinned in Vp intervention group [(0.93±0.27) μm, (4.99±1.75) μm,(26.59±2.76) μm, P<0.05]. The deposition proportion of collagen fiber around airway and PAS staining area decreased obviously(2.14%±0.89%, 17.92%±1.89%, P<0.05). The mRNA and protein expression levels of YAP and OPN decreased markedly (P<0.05 or P<0.01), and the ratio of p-YAP/YAP and the mRNA and protein expression levels of SMMHC increased (P<0.05). (2) Cell experiment: Compared with control group, the cell proliferation ability was higher (0.2874±0.0055 vs. 0.2626±0.0051, P<0.01)and apoptosis rate was lower in HDM group (5.14%±0.82% vs. 6.75%±0.21%, P<0.05). The protein expressions of YAP and OPN increased (3.14±0.48 vs. 1.51±0.61, 5.87±2.42 vs. 0.94±0.23, P<0.01), while of SMMHC decreased (0.80±0.19 vs. 2.96±0.96,P<0.01). Compared with HDM group, the cell proliferation ability reduced (0.2748±0.0043, P<0.05) and apoptosis rate increased(6.29%±0.49%, P<0.05), the protein expressions of YAP and OPN decreased (2.02±0.53, 2.93±1.09, P<0.05), while of SMMHC increased (2.11±0.85, P<0.05) in HDM+Vp group. Conclusions The abnormal expression of YAP may be involved in the airway remodeling in experimental asthma, and Vp may restrain the airway remodeling in experimental asthma by decreasing the YAP activity and nuclear shift, and reversing the HASMC phenotypic transformation.

Objective To screen the risk factors of preeclampsia and construct the predictive model of preeclampsia based on machine learning algorithm. Methods A retrospective study was conducted to collect the clinical data of 1609 hospitalized pregnant women from January 2016 to December 2018 on the big data platform of Academy of Medical Data Science of Chongqing Medical University. The 1609 cases were divided into preeclampsia group (n=291) and non-preeclampsia group (n=1318) according to the occurrence of preeclampsia during hospitalization. The clinical data of 70% patients were randomly selected as the training set (n=1126) to construct the prediction model, and the remaining 30% were used as the test set (n=483) for verification, and a consistency check between training set and test set was performed. The independent risk factors were screened by univariate analysis and logistic regression analysis, and the optimal parameters of LightGBM algorithm were searched by 5-fold cross-validation algorithm, and the prediction model was constructed based on LightGBM machine learning algorithm. Results A total of 58 indicators were collected, 13 indicators with missing rate ≥30% were excluded, and 45 indicators were finally included. Significant differences of 35 indicators existed between preeclampsia group and non-preeclampsia group (P<0.05) such as gamma-glutamyl transferase (GGT), alanine aminotrans ferase (ALT), thrombin time, aspartase transaminase (AST) and specific gravity of urine.Logistic regression analysis showed that specific gravity of urine, uric acid, hemoglobin concentration of erythrocyte, globulin, platelet distribution width, potassium ion, visiting age, family history of hypertension, systolic blood pressure, diastolic blood pressure,pulse and gestational age ≥34 weeks were independent risk factors for preeclampsia. The results of 5-fold cross-validation showed that, when num_leaves=5, max_depth=3, min_data_in_leaf=91, feature_fraction=0.8, bagging_fraction=0.6, and bagging_freq=5,the LightGBM model achieved the best effect the area under the curve (AUC), sensitivity and specificity of LightGBM model were 0.964, 84.9% and 92.7%. Conclusion The prediction model of preeclampsia based on LightGBM machine learning algorithm has a higher prediction effect, which can effectively predict the occurrence of preeclampsia in pregnant women in Chongqing, and provide decisions for clinicians.

Small cell lung cancer (SCLC) is an aggressive malignancy with a 5-year survival rate of 7.2%, which is characterized by exceptionally high proliferation rate, early metastasis, and poor prognosis. In the face of DNA damage and replicative stress, the DNA damage response (DDR) elicits activation of cell cycle checkpoints to promote repair by pausing the cell cycle or, in cases of unrepairable DNA damage, stimulate programmed cell death. SCLC carries a high mutation burden and genomic instability, and almost all SCLC tumors have functional inactivation of both TP53 and RB1, which leads to the form of a dysfunctional G₁/S checkpoint and results in an increased reliance on subsequent cell cycle checkpoints (S, G₂/M) to ensure genome stability and correct chromosomal segregation. Therefore, under the use of radio-chemotherapy for DNA damaging, promoting the cell cycle progress by inhibiting the cycle checkpoints and increasing DNA damage by inhibiting DNA damage repair can be new strategies for targeted therapies in SCLC. In the present article, we review the advances of targeting DDR and it's pathway inhibitors in SCLC, and provide some thoughts for the targeted treatment strategies of SCLC.

Vascular complications of type 2 diabetes mellitus (T2DM) include macrovascular complications and microvascular complications, of which the increased incidence is often accompanied with a high incidence of obesity. Obesity is an independent risk factor for T2DM. In obese patients, body fat distribution rather than total body fat is associated with metabolic and cardiovascular risk. Body mass index (BMI) and waist circumference (WC) cannot accurately reflect body fat distribution in obese patients, so visceral fat has received more and more attention and may be as a related risk factor for vascular complications in T2DM.The research progress on the relationship between visceral fat and vascular complications of T2DM and its action mechanism have been reviewed in present paper, in order to provide a reference for in-depth research and clinical examination of visceral fat.

Objective To observe the effect of iron overload on the cardiac function of mice and cardiac myocytes ferroptosis of rats, and explore the mechanism of action of iron transporter receptor (TFRC) in cardiac myocytes ferroptosis of rats. Methods Fourteen C57BL/6J mice were randomly divided into normal-iron diet (NID) group and high-iron diet (HID)group (7 each), fed with normal diet or high-iron diet for 8 weeks, respectively. The mice cardiac function was then detected by echocardiography; The content of malondialdehyde (MDA) in the heart was detected with ELISA; Masson staining was performed to detect the cardiac fibrosis; Propidium iodide (PI) staining and lipophilic fluorescent dye (C11) were used to detect the cell mortality and the content of lipid peroxide. The effect was observed of ferric ammonium citrate (AIC) and ferroptosis inhibitor ferrostatin-1 (Fer-1) on the cardiac myocytes of H9C2 rat. H9C2 cells were treated with AIC and ferroptosis inducer Erastin,Western blotting was performed to detect the expression level of TFRC and glutathione peroxidase 4 (GPX4) protein, qRT-PCR was used to detect the mRNA level; at the same condition, small interfering RNA was applied to knock-down the expression of TFRC,detect the mortality of H9C2 cells and the content of lipid peroxide. Results Compared with mice in NID group, the mice in HID group showed obviously increased interventricular septal thickness (IVSd), left ventricular posterior wall thickness (LVPWd), MDA relative content and area of collagen deposition (0.96±0.12 vs. 0.73±0.09, 1.18±0.28 vs. 0.84±0.07, 2.08±0.8 vs. 1.00±0.50,4.04±0.60 vs. 1.00±0.21, P<0.05); But no significant difference between the two groups on left ventricular ejection fraction and shortening fraction. Treated with 500 μmol/L AIC, the death rate and lipid peroxide content of H9C2 increased obviously(33.73%±1.20% vs. 2.30%±1.73%, 5.36±0.06 vs. 1.00±0.19, P<0.05), while after treatment with Fer-1, the death rate and lipid peroxide content of H9C2 decreased markedly (19.63%±0.81% vs. 33.73%±1.20%, 2.03±0.12 vs. 5.36±0.06, P<0.05). Treated with 500 μmol/L AIC, the expression level increased obviously of TFRC in H9C2 cell (P<0.05), while knocked-down of TFRC, the cell death rate and the content of lipid peroxide induced by high-iron decreased markedly (P<0.05). Under inducement of Erastin,the expression levels of TFRC and GPX4 protein in H9C2 cells obviously up- and down-regulated, respectively; and after knocked-down of TFRC, the cell death rate induced by Erastin decreased markedly (P<0.05). Conclusions HID may induce the damage of myocardial ferroptosis, and TFRC participated in the pathologic process induced by iron overload, which may be as a new target point for the treatment of heart diseases.

Objective To explore the mutation status of KRAS, NRAS, PIK3CA, and BRAF, and the relations with clinicopathological features and hormone expression levels in ovarian low-grade serous tumors. Methods Tissue specimens (23 cases of common-type serous borderline tumor, 27 cases of micropapillary serous borderline tumor and 14 cases of low-grade serous carcinoma) were collected from the First People's Hospital of Yunnan Province during January 2017 to April 2021, and then retrospectively analyzed. The mutation status of human KRAS, NRAS, PIK3CA and BRAF genes were detected by expression status in tumor tissues was detected by immunohistochemistry, the relationship was analyzed between gene mutation status and clinicopathological features and hormone expression levels. Results A total of 36 mutations were detected in 64 tumor tissues,including 21 KRAS mutation, 12 BRAF mutation, 2 PIK3CA mutation and 1 NRAS mutation. KRAS and BRAF mutations were common in younger women ≤35 years old. In the KRAS mutant subtype, G12S and G12D mutations are common in micropapillary serous borderline tumor (P<0.05); G12C, G12R*, G12V, G12A, and G13C* mutations are common in common-type serous borderline tumor (P<0.05); G13D mutation was only detected in low-grade serous carcinoma. Patients with BRAF mutations were all seen in the early stages of the disease. Immunohistochemistry showed the positive expression rates of both estrogen receptor and progesterone receptor were 76.6% (49/64). The positive expression rate of common-type serous borderline tumor was similar to that of micropapillary serous borderline tumor, and both of them were significantly higher than that in lower-grade serous carcinoma(P<0.05). KRAS G13D mutation was significantly associated with low progesterone expression (P<0.05). Conclusion Gene mutations were mutual exclusion. KRAS mutation was an oncogenic driving factor in serous tumors, and different mutant subtypes were associated with histological type and progesterone receptor expression status. As the disease progresses, BRAF mutation rate gradually decreased and associated with a variety of clinicopathological features and good prognosis and might have a protective effect against disease progression. The mutation rates of NRAS and PIK3CA were lower.

Objective To prepare fibrin-targeted, integration of diagnosis and treatment and multi-functional gelatin nanoparticles, explore their potential of dual-mode imaging and the mechanism and effect of photothermal synergistic drug thrombolytic therapy in vitro, and observe the active-targeting ability of thrombus in vivo. Methods Loaded with alteplase (rt-PA)and Fe₃O₄ mediated by targeting fibrin polypeptide (Cys-Arg-Glu-Lys-Ala, CREKA), the magnetic gelatin nanoparticles (CREKA-Fe-rt-PA-Gel) were prepared by two-step desolvation and carbodiimide method. The morphology of CREKA-Fe-rt-PA-Gel was observed, and its physical and chemical properties and in vitro safety were detected. Its photoacoustic (PA) and magnetic resonance(MR) imaging features were also verified in vitro. The photothermal performance was monitored, the fibrin and hemoglobin indexes were used to evaluate the thrombolytic effect of photothermal therapy and the ability to damage fibrin network. Carotid artery thrombosis model was constructed with rats and HE staining section was performed to observe the targeting thrombus ability in vivo of the nanoparticles. Results CREKA-Fe-rt-PA-Gel were with uniform particle size, diameter of (254.79±6.83) nm, polydispersity coefficient of 0.09±0.05, surface potential of (5.48±4.60) mV. The CREKA peptide was successfully connected with the target connection rate of 98.66%, the encapsulation efficiency of rt-PA was 23.03%±0.05%, and of Fe₃O₄ was 92.78%±0.57%, the magnetization curve showed that it has good paramagnetism. The cytotoxicity and hemolysis test in vitro confirmed that CREKA-Fe-rt-PA-Gel had appropriate biological compatibility. At the same time, CREKA-Fe-rt-PA-Gel showed excellent imaging ability of PA and MR imaging in vitro. After laser irradiation, the nano solution heated up rapidly in a concentration-dependent manner,hemoglobin and fibrin in solution increased significantly after incubation with thrombus by near infrared irradiation (P<0.01), and nanoparticles released drugs combined with photothermal therapy could damage the fibrin network. HE staining of carotid artery thrombosis in rats showed good thrombus targeting ability of the nanoparticles in vivo. Conclusion The constructed CREKA-Fe-rt-PA-Gel have potential in dual-mode (PA/MR) imaging, good photothermal properties, outstanding targeting ability for thrombus(fibrin), and excellent thrombolytic ability when combined with photothermal, which is expected to be used in early diagnosis and multi-form treatment in clinical thrombotic diseases.