Anti-tumor efficacy of HRS-4642 and its potential combination with proteasome inhibition in KRAS G12D-mutant cancer.

KRAS G12D is the most frequently mutated oncogenic KRAS subtype in solid tumors and remains undruggable in clinical settings. Here, we developed a high affinity, selective, long-acting, and non-covalent KRAS G12D inhibitor, HRS-4642, with an affinity constant of 0.083 nM. HRS-4642 demonstrated robust efficacy against KRAS G12D-mutant cancers both in vitro and in vivo. Importantly, in a phase 1 clinical trial, HRS-4642 exhibited promising anti-tumor activity in the escalating dosing cohorts. Furthermore, the sensitization and resistance spectrum for HRS-4642 was deciphered through genome-wide CRISPR-Cas9 screening, which unveiled proteasome as a sensitization target. We further observed that the proteasome inhibitor, carfilzomib, improved the anti-tumor efficacy of HRS-4642. Additionally, HRS-4642, either as a single agent or in combination with carfilzomib, reshaped the tumor microenvironment toward an immune-permissive one. In summary, this study provides potential therapies for patients with KRAS G12D-mutant cancers, for whom effective treatments are currently lacking.

Evaluation on the Risk of Water Inrush Due to Roof Bed Separation Based on Improved Set Pair Analysis-Variable Fuzzy Sets.

To study and prevent bed separation water inrush accidents in coal mines, it is necessary to evaluate the risk according to the limited geological data correctly. In this work, based on hydrogeological and mining conditions, we established a risk evaluation model and selected seven important factors, including the aquifer thickness, aquifer water abundance, hydraulic pressure of the aquifer, effective aquifuge thickness, mining failure ratio, mining height of the working face, and advancing distance as evaluation indexes. The intuitionistic fuzzy analytic hierarchy process (IFAHP) and entropy weight method (EWM) were used to analyze the weights of the original data, and the minimum information entropy principle was used to further integrate the abovementioned calculation results. With the weight results, set pair analysis-variable fuzzy set (SPA-VFS) theory was applied to determine the risk grade of each working face, which provided scientific guidance for the safe mining of coal mines. For the working face where water inrush may occur, the risk of bed separation water inrush can be reduced by optimizing the parameters or changing the mining conditions through the model analysis.

Mathematical Evaluation on the Control of Mining-Induced Ground Subsidence in Thick Loose Strata.

Coal mining under thick loose strata in North China leads to ground subsidence, which is a natural result of hydromechanical coupling (fluid flow coupled with solid deformation). Therefore, the land surrounding the mining areas is greatly damaged. In this study, the combined weight (CW) method and the fuzzy matter-element analysis (FMEA) method were used to analyze and evaluate the control effect of subsiding land. Overall, 20 sets of geological samples were collected from this area. The influencing factors and the corresponding weights for the control effect of subsiding land were comprehensively analyzed, and an FMEA model was built to predict and verify the results. The results showed that (1) the two evaluation indicators with the most significant impact on land reclamation were the degree of integration and the economic and social benefits and (2) among the 20 sets of samples selected, the predicted conclusions of 17 sets were consistent with the actual engineering situations, which led to an accuracy of 85%. In other words, the CW-FMEA model showed good reliability for evaluating the control effect of subsiding land, which can provide scientific references for control and quality evaluations of land subsidence due to coal mining.

Correction to "New Type of Similar Material for Simulating the Processes of Water Inrush from Roof Bed Separation".

[This corrects the article DOI: 10.1021/acsomega.0c03535.].

New Type of Similar Material for Simulating the Processes of Water Inrush from Roof Bed Separation.

With an increase in the mining depth and breadth of coal mines in China, water inrush accidents from the roof bed separation of a mining face have become more frequent. A similar simulation experiment in the laboratory is an important way to study the dynamic seepage processes of separation water and explore the mechanism of roof bed separation formation. In this study, we develop a new type of similar material to simulate strata separation. The skeleton of this material is made from river sand, while nanosized calcium carbonate, Ca-bentonite, gypsum, and emulsified wax act as additives. These synthetic materials are then used for the compressive strength experiment, the water permeation height experiment, the structural stability experiment, and the microscopic analysis. Besides, a physical experiment is also conducted to verify the effectiveness of this new similar material. This work could provide the scientific basis for the prediction and control of water accidents caused by the separation strata.

Experimental Study on the Basic Properties of a Green New Coal Mine Grouting Reinforcement Material.

Waste tires are internationally recognized as hazardous waste. Many waste tires storing in the open air for a long time will not only waste the land resources but also pollute the environment after the incineration. Meanwhile, the cumulative accumulation of fly ash in China has also been one of the largest sources of solid waste in coal mines. Under the action of high winds, the accumulated fly ash forms flying dust, which pollutes the environment and endangers human health. Herein, a new type of grouting material for floor reinforcement was developed based on solid waste products using the waste tire rubber particles and fly ash. Using this new material, disaster accidents such as flooding and casualties caused by the inrush of the floor-confined water due to floor fractures, activation, and expansion in the mining process can be greatly reduced, thereby ensuring the safety of coal mine production. The grout body was prepared using a large amount of fly ash, waste tire rubber particles and clay, and mixing additives. The ratio optimization test, uniaxial compressive strength test, permeability characteristic test, stability test, and microanalysis of mine water environment were performed. From the test results, the macrophysical and chemical properties including optimal mix ratio, compressive strength, permeability coefficient, and stability of mine water environment were obtained. The microstructural properties of the grouts were analyzed using scanning electron microscopy microanalysis methods. Considering the situation of the coal floor and attempting to use as many solid waste products as possible, the optimized proportion that can meet the requirements of low cost, high bonds, and dense filling is as follows: 20% of rubber particles, 65% of fly ash, 15% of clay, and 1% of nanosilica. Furthermore, this study can provide scientific reference for large-scale floor grouting reinforcement and large-scale utilization of solid waste products.

[Analysis of primary site and pathology on 903 patients with neuroendocrine neoplasms].

OBJECTIVE: To explore the primary site and pathological feature of neuroendocrine neoplasm (NEN), especially the NEN of digestive system. METHODS: Clinicopathological data of NEN patients at China-Japan Friendship Hospital from January 2012 to December 2016 were retrospectively analyzed. Tumor primary sites were summarized. Association between tumor site and pathological grading in gastroenteropancreatic neuroendocrine neoplasm(GEP-NEN) was examined. RESULTS: There were a total of 903 cases of NEN. Sites of primary tumor included the digestive system in 699 cases(77.4%), the thorax(including lung, thymus and mediastinum) in 87 cases(9.6%), other sites in 60 cases (6.6%), unknown in 57 cases(6.3%). Among 699 GEP-NEN cases, the primary sites included the stomachin in 207 cases (29.6%), pancreas in 201 (28.8%), rectumin in 185 (26.5%), duodenum in 43(6.2%), jejunum and ileum in 18(2.6%), appendix in 15 (2.1%), gallbladder in 11(1.6%), esophagus in 10(1.4%), and the colon in 9 cases (1.3%). Pathologically, the tumor grading was neuroendocrine tumor (NET) G1 in 336 cases(48.1%), NET G2 in 203 cases (29.0%), neuroendocrine carcinoma (NEC) G3 in 139 cases (19.9%). All the esophagus NEN(10/10), most gallbladder NEN(9/11) and colon NEN(6/9) were poorly-differentiated NEC (G3), while all appendix NEN(15/15), most stomach NEN(147/207, 71.0%), pancreas NEN (156/201, 77.6%), rectum NEN (169/185, 91.4%), duodenum NEN (31/43, 72.1%), jejunum and ileum NEN(16/18, 88.9%) were well-differentiated NET G1 or G2. CONCLUSIONS: The most common primary site of NEN is the digestive system. The stomach, pancreas and rectum are most common primary sitesof GEP-NEN. Difference in pathological grading is quite greatin different primary sites of GEP-NEN. Most NENs fromesophagus, colon and gallbladder are poorly-differentiated NEC.

Down-regulation of microRNA-144 in air pollution-related lung cancer.

Air pollution has been classified as a group 1 carcinogen in humans, but the underlying tumourigenic mechanisms remain unclear. In Xuanwei city of Yunnan Province, the lung cancer incidence is among the highest in China, owing to severe air pollution generated by the combustion of smoky coal, providing a unique opportunity to dissect lung carcinogenesis. To identify abnormal miRNAs critical for air pollution-related tumourigenesis, we performed microRNA microarray analysis in 6 Xuanwei non-small cell lung cancers (NSCLCs) and 4 NSCLCs from control regions where smoky coal was not used. We found 13 down-regulated and 2 up-regulated miRNAs in Xuanwei NSCLCs. Among them, miR-144 was one of the most significantly down-regulated miRNAs. The expanded experiments showed that miR-144 was down-regulated in 45/51 (88.2%) Xuanwei NSCLCs and 34/54 (63%) control region NSCLCs (p = 0.016). MiR-144 interacted with the oncogene Zeb1 at 2 sites in its 3' untranslated region, and a decrease in miR-144 resulted in increased Zeb1 expression and an epithelial mesenchymal transition phenotype. Ectopic expression of miR-144 suppressed NSCLCs in vitro and in vivo by targeting Zeb1. These results indicate that down-regulation of miR-144 is critical for air pollution-related lung cancer, and the miR-144-Zeb1 signalling pathway could represent a potential therapeutic target.

Tobacco smoke induces production of chemokine CCL20 to promote lung cancer.

Tobacco kills nearly 6 million people each year, and 90% of the annual 1.59 million lung cancer deaths worldwide are caused by cigarette smoke. Clinically, a long latency is required for individuals to develop lung cancer since they were first exposed to smoking. In this study, we aimed to identify clinical relevant inflammatory factors that are critical for carcinogenesis by treating normal human lung epithelial cells with tobacco carcinogen nicotine-derived nitrosaminoketone (NNK) for a long period (60 days) and systematic screening in 84 cytokines/chemokines. We found that a chemokine CCL20 was significantly up-regulated by NNK, and in 78/173 (45.1%) patients the expression of CCL20 was higher in tumor samples than their adjacent normal lung tissues. Interestingly, CCL20 was up-regulated in 48/92 (52.2%) smoker and 29/78 (37.2%) nonsmoker patients (p = 0.05), and high CCL20 was associated with poor prognosis. NNK induced the production of CCL20, which promoted lung cancer cell proliferation and migration. In addition, an anti-inflammation drug, dexamethasone, inhibited NNK-induced CCL20 production and suppressed lung cancer in vitro and in vivo. These results indicate that CCL20 is crucial for tobacco smoke-caused lung cancer, and anti-CCL20 could be a rational approach to fight against this deadly disease.

Quercetin prevents pyrrolizidine alkaloid clivorine-induced liver injury in mice by elevating body defense capacity.

Quercetin is a plant-derived flavonoid that is widely distributed in nature. The present study is designed to analyze the underlying mechanism in the protection of quercetin against pyrrolizidine alkaloid clivorine-induced acute liver injury in vivo. Serum transaminases, total bilirubin analysis, and liver histological evaluation demonstrated the protection of quercetin against clivorine-induced liver injury. Terminal dUTP nick end-labeling assay demonstrated that quercetin reduced the increased amount of liver apoptotic cells induced by clivorine. Western-blot analysis of caspase-3 showed that quercetin inhibited the cleaved activation of caspase-3 induced by clivorine. Results also showed that quercetin reduced the increase in liver glutathione and lipid peroxidative product malondialdehyde induced by clivorine. Quercetin reduced the enhanced liver immunohistochemical staining for 4-hydroxynonenal induced by clivorine. Results of the Mouse Stress and Toxicity PathwayFinder RT2 Profiler PCR Array demonstrated that the expression of genes related with oxidative or metabolic stress and heat shock was obviously altered after quercetin treatment. Some of the alterations were confirmed by real-time PCR. Our results demonstrated that quercetin prevents clivorine-induced acute liver injury in vivo by inhibiting apoptotic cell death and ameliorating oxidative stress injury. This protection may be caused by the elevation of the body defense capacity induced by quercetin.

Proteomic characterization of the possible molecular targets of pyrrolizidine alkaloid isoline-induced hepatotoxicity.

Pyrrolizidine alkaloids (PAs) are distributed in plants worldwide including medicinal herbs or teas. In the present study, we investigated the effects of isoline, which is a retronecine-type PA isolated from traditional Chinese medicinal herb Ligularia duciformis, on mouse liver proteins by using proteomic approaches. Firstly, our results showed that 110mg/kg isoline increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum, and hepatic tissue pathological observation further confirmed isoline-induced liver injury. Proteomic analysis showed that the liver samples from mice of isoline group demonstrated about 13 differentially expressed proteins compared with normal group, and those proteins may be involved in isoline-induced liver injury in mice. Next, all these 13 protein spots were identified by MALDI-TOF-TOF MS or LTQ MS; and among them 9 differentially expressed proteins are involved in the process of oxidative stress or cellular energy metabolism. Further lipid peroxidation analysis and ATPase assay confirmed the existing of oxidative injury induced by isoline and consequent disruption of energy metabolism. Furthermore, an in silico drug target searching program INVDOCK identified 2 potential protein targets of isoline, and the results are in support of proteomic analysis. In summary, the possible signaling molecules related with isoline-induced liver injury were demonstrated in this study.

[Primary safety evaluation of sulfated paeoniae radix alba].

The paper is to report the development of a method of quantitative analysis of multi-components by high performance liquid chromatography (HPLC) for simultaneously determining paeoniflorin sulfonate (PS), paeoniflorin (PF) and albiflorin (AF) in sulfated Paeoniae Radix Alba. Moreover, the cytotoxicity of paeoniflorin sulfonate by MTT-assay and the acute toxicity of mice by administration of paeoniflorin sulfonate were evaluated. Chromatographic separation of paeoniflorin sulfonate, PF and AF were performed on a SHISEIDO CAPCELL PAK C18 column (250 mm x 4.6 mm, 5 microm) for HPLC and a mixture of acetonitrile and 0.02% phosphoric acid solution (15 : 85) as the mobile phase. As detector a spectrophotometer set at 230 nm; column temperature 30 degrees C; flow rate 1.0 mL x min(-1). The toxicity of paeoniflorin sulfonate was evaluated by in vitro cytotoxicity carried out on mouse and human primary hepatocytes, and by acute oral toxicity test carried out on mice. The calibration curve of paeoniflorin sulfonate, PF and AF revealed linearity in the range of 0.041 8 - 1.045 0, 0.023 5 - 0.587 5, and 0.039 8 - 0.995 0 mg x mL(-1), respectively (r > 0.999 8). The average recovery was ranged from 99.11% to 101.71%, RSD < 2%. Paeoniflorin sulfonate does not have any cytotoxicity to cells at all the tested concentrations (< or = 300 micromol x L(-1)) in the in vitro cytotoxicity assay. The maximum tolerance dose of paeoniflorin sulfonate solution and extraction of Paeoniae Radix Alba to mouse is 5 g x kg(-1) and 80 g x kg(-1) respectively. The contents of these three components in the samples were determined with the developed method. It is a rapid, convenient and accurate method to determine multi-components. The content of PF in sulfated Paeoniae Radix Alba is significantly lower, and there is negative correlationship between the content of paeoniflorin sulfonate and PF. The in vitro cytotoxicity assay and in vivo mouse acute toxicity test showed that there is no obvious toxicity of paeoniflorin sulfonate and water-soluble extract of sulfated Paeoniae Radix Alba.

Antitumor activity of Dioscorea bulbifera L. rhizome in vivo.

Antitumor activities of water extract (fraction A), ethanol extract (fraction B), ethyl acetate extract (fraction C), non-ethyl acetate extract (fraction D) and compound diosbulbin B isolated from Dioscorea bulbifera L. (DB) were investigated in vivo in this present study. The results showed that fractions B and C both decreased tumor weight in S180 and H22 tumor cells bearing mice, while fractions A and D had no such effect. Furthermore, fraction C altered the weight of spleen and thymus, and the amounts of total leukocytes, lymphocytes and neutrophils in tumor-bearing mice. Further results showed that compound diosbulbin B demonstrated anti-tumor effects in the dose-dependent manner at the dosage of 2 to 16 mg/kg without significant toxicity in vivo. Furthermore, on the basis of chemical analysis of the above extracts by high-performance liquid chromatography (HPLC) with a diode array detector (DAD), diosbulbin B was found to be the major antitumor bioactive component of DB. These results suggest that DB has potential anti-tumor effects which may be related to influencing the immune system for the first time, and the compound diosbulbin B is the major antitumor component of DB.

Pyrrolizidine alkaloid isoline-induced oxidative injury in various mouse tissues.

Isoline is a retronecine-type pyrrolizidine alkaloid (PA) isolated from the traditional Chinese medicinal herb Ligularia duciformis. The present investigation was carried out to evaluate isoline-induced oxidative injury in various important mouse organs. Various tissue samples were collected after mice were administrated with 100mg/kg isoline for 36h, and then lipid peroxidation (LPO) level, total antioxidant capacity, glutathione-S-transferase (GST), glutathione peroxidase (GPx) and catalase (CAT) activities were determined to evaluate the oxidative injury. Our results showed that the total antioxidant capacity of liver, brain and lung were all decreased after given isoline, and the LPO level was increased in liver and heart of isoline-treated mice. Further antioxidant-related enzyme activity assays showed that isoline (100mg/kg) decreased GPx activity in liver and heart, increased CAT activity in liver, brain and heart, and decreased the GST activity in lung. Taken together, our results demonstrate that isoline can induce different oxidative injury in various important mouse organs, and of which liver is the most sensitive organ.

[The cryosurgery of central lung cancer by rigid bronchoscopy].

OBJECTIVE: To evaluate the method and effectiveness of rigid-bronchoscopic cryosurgery for advanced central lung cancer. METHODS: Forty-eight patients were enrolled in this study from June 2002 to December 2008, including 33 male and 15 female. The average age was 70 years (ranged from 45 to 83 years old). For the 48 patients, 38 cases were patients with advanced central lung cancer who were not suitable for surgery, and the remaining 10 cases were patients with local recurrence in trachea or main bronchus postoperatively. Cryosurgery was performed 120 times for all patients, 2.5 times per patient on average. The trachea or bronchus station, symptom such as dyspnea, hemoptysis, respiratory function and quality of life were observed. RESULTS: The unblocked ratio of trachea and bronchi was 97%. All patients got satisfied improvement ratio of symptoms, 87.5% for dyspnea, 72.9% for cough, 93.8% for hemoptysis and 62.5% for chest pain. Respiratory function tests showed that both the mean forced expiratory volume in first second and forced vital capacity got an improvement from (1.03+/-0.05) L to (1.85+/-0.13) L and from (1.69+/-0.18) L to (2.96+/-0.14) L respectively (P<0.01). Karnofsky score also got no less than 20 scores improvement. The Follow-up time was 6 to 62 months. The longest survival was 62 months. The median survival time was 20 months. There was no severe perioperative complications and mortality except for 3 cases of moderate exeduation. CONCLUSIONS: Cryosurgery is easy to perform with minimal complications. Not only could it provide an effective and rapid control of symptoms caused by central lung cancer, it could also unobstructed bronchus promptly and improve patients' quality of life.

Bile acids metabonomic study on the CCl4- and alpha-naphthylisothiocyanate-induced animal models: quantitative analysis of 22 bile acids by ultraperformance liquid chromatography-mass spectrometry.

Bile acids (BAs) are crucial for the diagnosis, follow-up, and prognostics of liver and intestinal disorders and other diseases affecting BA metabolism. A rapid, simple, and sensitive analytical method is needed to demonstrate the full metabolic profile and simultaneously determine the individual BAs in biological samples. In our present study, an ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) method has been established and validated for simultaneous quantitation of 22 BAs and a metabonomic study was performed based on the chemometric analysis of the serum samples from carbon tetrachloride (CCl4)- and alpha-naphthylisothiocyanate (ANIT)-induced liver failure rats. The optimal chromatographic condition was effected by UPLC (Acquity UPLC BEH column, 1.7 microm, 2.1 mm x 100 mm) using a linear gradient elution system of methanol-5 mM ammonium acetate containing 0.01% acetic acid after a simple-step deproteinization by precipitation. The separation of the 22 BAs can be finished in less than 12 min, and the concentrations of these BAs in rat serums were simultaneously determined using a selective ions monitoring mode. The method was validated with respect to repeatability (relative standard deviation < 9.78%) and accuracy (relative errors from -13.55 to 9.58%). The range of each BA was found from not detected (nd) to 8301 ng mL(-1), respectively. Furthermore, the developed method was successfully applied to the metabonomics analysis of BAs in CCl4- and ANIT-induced liver failure rats, using principle component analysis and canonical discriminant analysis. The serum samples from the two types of rat liver injury could be distinguished from each other and from the untreated animals according to the varieties of BAs. It indicated that the level of BAs could be considered as a sensitive parameter of hepatotoxicity induced by different chemical toxins. This novel metabonomics study of BAs based on the UPLC-MS profile provides not only an accurate quantitative assay of the serum concentrations of biomarkers but also a promising methodology for evaluation of liver injury.

[Results of unilateral lung volume reduction surgery in twenty-five patients with chronic obstructive pulmonary disease].

OBJECTIVE: To evaluate the effectiveness of unilateral lung volume reduction surgery (LVRS) in patients with chronic obstructive pulmonary disease (COPD). METHODS: The follow-up data of 25 patients with COPD who had underwent unilateral LVRS between January 1996 to December 2002 in department of thoracic surgery, China-Japan friendship hospital were analyzed retrospectively. The operative target was determined by pre-operative CT and pulmonary ventilation-perfusion (V/Q) scintigraphy. LVRS was performed in 21 patients through video assisted thoracoscopy surgery (VATS) or VATS with adjuvant small lateral thoracotomy. In 4 patients LVRS was performed through posterolateral thoracotomy. Destroyed pulmonary tissue was resected by liner stapler or Endo GIA. To evaluate the effectiveness of unilateral LVRS, the changing of dyspnea score, pulmonary function and the quality of life were analyzed. RESULTS: The postoperative follow-up interval range was 2 years. The mean postoperative FEV(1) increased by (35 +/- 9)%, and six minute walking distance (6MWD) increased by (88 +/- 22)%. For dyspnea score, among 16 patients with a preoperative grade of IV, 4 patients improved to grade I, 12 improved to grade II; among 9 patients with a preoperative grade of V, 1 improved to grade I, 1 improved to grade II, 4 improved to grade III, and the other 3 improved to grade IV. Karnofsky score increased by (44 +/- 10) in average. One and two year survival rates was 96% and 92%, respectively. There was no perioperative death in this group, and the total postoperative morbidity was 32%. CONCLUSIONS: Unilateral LVRS shows significant clinical benefits for the majority of patients with COPD. It is associated with lower operative mortality and morbidity, and has a wide range of indications. The key points of the operation are to resect dysfunctional lung tissues as much as possible and to prevent pulmonary air leak. Preoperative and postoperative breathing training plays a very important role in the postoperative recovering of lung function.

[Late-stage emphysema treated with lung volume reduction: report of 22 cases].

OBJECTIVE: To Summarize the clinical experience in the treatment of late-stage emphysema by lung volume reduction (LVR) in 5 years. METHODS: We retrospectively studied the indications, contraindications, operation procedures and complications of LVR in 22 patients. RESULTS: Before operation, the average FEV(1) was 24.5%, RV 196.8%, and TLC 130.5%; after operation they were 27.8%, 148.8% and 112.5%, respectively. 16 patients needed inhaling oxygen before operation, and 5 after operation. 16 patients finished 6-minute walking test with an average of 198 m, all patients walked much longer with an average of 256 m after operation. 3-degree lung function was observed in 14 patients, and 4-degree before operation in 8 patients; but 2-degree lung function in 5 patients, 3-degree in 13, and 4-degree in 4 after operation. CONCLUSIONS: Heterogeneous type emphysema with clear target area, especially bullous emphysema is the best indication for LVR. Lung function and life quality could be much improved postoperatively. Homogeneous type could also be treated with LVR in highly selected cases. TLCO < 20% is not an absolute contraindication, others standards need further investigation. Video-assistant thoracoscopic surgery (VATS) with subaxillary small incision for LVR is safe, reliable and effective. Application of stapler buttressing with bovine pericardia could decrease air leakage postoperatively.