2. Expression, purification, and functional verification of recombinant human glycoprotein hormone beta 5/alpha 2 fusion protein in CHO-S cells
Ai-Jun QIAN ; Geng-Miao XIAO ; Zhuang LI ; Yun-Ping MU ; Zi-Jian ZHAO ; Fang-Hong LI ; Zhi-Cheng LIANG
Chinese Pharmacological Bulletin 2024;40(2):390-396
		                        		
		                        			
		                        			 Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice. 
		                        		
		                        		
		                        		
		                        	
3.Clinical Experience of LI Yan in Treating Coronary Heart Disease from the Spleen and Stomach
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(1):213-218
		                        		
		                        			
		                        			Coronary heart disease(CHD)is a common cardiovascular disease in clinic,which can be classified as"chest qi-obstruction"and"heart pain"in traditional Chinese medicine.Professor LI Yan believes that CHD can be differentiated as the syndrome of deficiency in origin and excess in superficiality,and the deficiency in origin is characterized by the deficiency of spleen and the failure of spleen and stomach in transportation,while the excess in superficiality manifests as the obstruction of visible pathogens such as phlegm turbidity,blood stasis,qi stagnation,and cold accumulation.The clinical treatment should focus on regulating the spleen and stomach,using Astragali Radix,Atractylodis Macrocephalae Rhizoma,Codonopsis Radix,Schisandrae Chinensis Fructus to strengthen the spleen and nourish the heart in order to treat the root cause.Moreover,therapies of activating yang to remove turbidity,activating blood to remove stasis,soothing liver to rectify qi,and warming yang to dissipate the cold can be adopted.Therefore,the symptom and root cause will be treated simultaneously,the unblocking and supplementing effect will be achieved,and then the spleen-qi is vigorous,the heart qi is in abundance,the blood stasis and the phlegm are removed,and the chest yang can be stretched.As a result,the heart vessels are unblocked,and the symptoms of chest qi-obstruction will be relieved.Professor LI Yan's experience from the perspective of spleen and stomach can provide reference for the clinical diagnosis and treatment of CHD with traditional Chinese medicine.
		                        		
		                        		
		                        		
		                        	
4.Impact of early nasogastric tube removal on functional recovery after total gastrectomy
Ran HU ; Zhi ZHENG ; Yuhao ZHAI ; Lihong ZHAO ; Dan XUE ; Chenchen SONG ; Jun ZHANG ; Meng SUN
International Journal of Surgery 2024;51(8):542-547
		                        		
		                        			
		                        			Objective:To investigate the impact of early removal of nasogastric tubes on functional recovery after total gastrectomy for gastric cancer patients, to provide scientific evidence for enhanced recovery after surgery strategies in gastric cancer.Methods:A retrospective cohort study was conducted on 102 gastric cancer patients who underwent total gastrectomy at Beijing Friendship Hospital affiliated with the Capital Medical University from March 2018 to July 2022. Patients were divided into two groups based on whether the gastric tube was removed within 24 hours post-operation: the early removal group (within 24 hours, 55 patients) and the non-early removal group (beyond 24 hours, 47 patients). The recovery outcomes, including time to first flatus, time to fluid intake, length of hospital stay, and the incidence of postoperative complications, were compared between the two groups. Non-normally distributed data were expressed as M( Q1, Q3) and compared using the Wilcoxon rank-sum test. Categorical data were expressed as frequencies or percentages and compared using the chi-square test or Fisher′s exact test. To minimize the impact of potential confounders, multivariable linear regression and logistic regression analyses adjusted for propensity scores were further employed to compare quantitative and qualitative data between the groups. Statistical analyses were performed using R software. Results:The exhaust time, water intake time, and hospital stay in the early removal group were 3.0(2.0, 3.5) days, 4.0(3.0, 5.0) days, and 7.0(7.0, 8.0) days, respectively, while those in the non-early removal group were 4.0(3.0, 5.0) days, 6.0(5.0, 7.0) days, and 8.0(7.5, 11.0) days, respectively. Statistically significant differences were observed between the two groups ( P<0.05). However, there was no significant difference in the incidence of postoperative complications between the two groups (5.45% vs 17.02%, P=0.060). Propensity score-adjusted regression analysis suggested that early tube removal might reduce the risk of postoperative complications ( P=0.042). Conclusion:Early removal of nasogastric tubes can significantly accelerate functional recovery after total gastrectomy for gastric cancer patients and may reduce the risk of postoperative complications, providing important clinical guidance for enhanced recovery after surgery management in gastric cancer.
		                        		
		                        		
		                        		
		                        	
5.Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines
Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG
Parasites, Hosts and Diseases 2024;62(4):476-483
		                        		
		                        			
		                        			 Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines. 
		                        		
		                        		
		                        		
		                        	
6.Comparison of the efficacy of TiRobot orthopaedic robot assisted F screw technique and inverted triangle parallel nail internal fixation in the treatment of unstable femoral neck fractures
Xing-Long ZHAO ; Jian-Jun SHEN ; Kang-Hu FENG ; Zhi-Wei CHEN ; Yuan-Long SI ; Xuan ZHANG ; Guan-De WANG ; Xiang HAI
China Journal of Orthopaedics and Traumatology 2024;37(2):129-134
		                        		
		                        			
		                        			Objective To compare the effectiveness of TiRobot assisted F screw technique and inverted triangle parallel nail internal fixation in the treatment of unstable femoral neck fractures.Methods A retrospective analysis was conducted on 72 patients with unstable femoral neck fractures who were treated with percutaneous cannulated screw fixation assisted with TiRobot Orthopaedic robot from December 2019 to April 2021.Among them,37 patients were treated with F screw internal fixa-tion,including 16 males and 21 females,aged47 to 64years old with an average of(53.87±5.28)years old;According to Pauwels classification,there were 1 case of type Ⅰ,19 cases of type Ⅱ,17 cases of type Ⅲ;8 cases of combined medical diseases;17 cases of falling,8 cases of traffic accident and 12 cases of falling from height;The time from injury to operation was 29 to 49 hours with average of(35.00±7.34)hours.Another 35 cases used internal fixation with an inverted triangle parallel nail,including 13 males and 22 females with an average age of 46 to 63 years old(52.36±5.05)years old;According to the Pauwels injury classifi-cation:there were 2 cases of type Ⅰ,21 cases of type Ⅱ,12 cases of type Ⅲ;6 cases of medical diseases,15 cases of falling in-jury,9 cases of traffic accident,11 cases of falling injury;The time from injury to operation was 30 to 45 hours with an average of(33.00±6.83)h.The intraoperative blood loss,operation time,intraoperative fluoroscopy times,follow-up time,fracture healing time,postoperative complications were observed and compared between the two groups.The hip joint function was e-valuated by Harris score at 6 months and 12 months after operation.Results There was no significant difference in operation time,intraoperative blood loss,intraoperative fluoroscopy times and other intraoperative data between two groups(P>0.05).Both groups were followed up regularly,and the follow-up time was 12 to 16 months.The fracture healing time and Harris score of the F screw internal fixation group were better than those of the inverted triangle parallel nail internal fixation group(P<0.05).There was 1 case of femoral neck shortening in the F screw internal fixation group,1 case of nonunion,1 case of nail withdrawal,and 1 case of lower extremity deep vein thrombosis in the inverted triangle internal fixation group.The incidence of complications in the F screw internal fixation group was lower than that in the inverted triangle parallel nail internal fixation group(P<0.05).Conclusion Percutaneous cannulated F screw technique using Tirobot navigation positioning system is a safe and effective treatment for patients with unstable femoral neck fractures.It can significantly shorten the fracture healing time,reduce the incidence of postoperative complications,significantly improve hip joint function,and improve the quality of life.
		                        		
		                        		
		                        		
		                        	
7.Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines
Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG
Parasites, Hosts and Diseases 2024;62(4):476-483
		                        		
		                        			
		                        			 Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines. 
		                        		
		                        		
		                        		
		                        	
8.Chinese clinical guidelines on diagnosis and treatment of lung cancer bone metastasis (version 2024)
Xiuyi ZHI ; Jie WANG ; Lunxu LIU ; Jun ZHAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(05):643-653
		                        		
		                        			
		                        			Lung cancer is the highest morbidity of malignant tumor in China, and bone metastasis is one of the common sites. With the development of imaging and nuclear medicine technology, the level of early diagnosis of bone metastasis has been improved. There are also many evidence-based evidences and advances in systemic therapy (chemotherapy, targeted therapy, immunotherapy) and bone modification drugs for treatment of bone metastases from lung cancer. The comprehensive treatment model under the guidance of multiple disciplines (including medical oncology, surgery, radiotherapy, interventional medicine, nuclear medicine, psychological rehabilitation, etc.) has been widely implemented in clinical practice. Therefore, Lung Cancer Medical Education Committee of China Medicine Education Association, Youth Specialists Committee of Lung Cancer, Beijing Medical Award Foundation and Lung Cancer Specialty Committee of Chinese Elderly Health Care Association have written the "Chinese Clinical Guidelines on Diagnosis and Treatment of Lung Cancer Bone Metastasis (Version 2024)", based on the "Expert Consensus on Diagnosis and Treatment of Lung Cancer Bone Metastasis (Version 2019)". The aim is to enhance the comprehensive treatment level of lung cancer bone metastasis in China.
		                        		
		                        		
		                        		
		                        	
9.Application of CRISPR/Cas-based Electrochemical Biosensors for Tumor Detection
Shuang LI ; Zhi CHEN ; Yun-Xia HUANG ; Guo-Jun ZHAO ; Ting JIANG
Progress in Biochemistry and Biophysics 2024;51(8):1771-1787
		                        		
		                        			
		                        			Tumors represent one of the primary threats to human life, with the dissemination of malignant tumors being a leading cause of mortality among cancer patients. Early diagnosis of tumors can reliably predict their progression, significantly reducing mortality rates. Tumor markers, including circulating tumor cells, exosomes, proteins, circulating tumor DNA, miRNAs and so on, generated during the tumor development process, have emerged as effective approach for early tumor diagnosis. Several methods are currently employed to detect tumor markers, such as polymerase chain reaction, Northern blotting, next-generation sequencing, flow cytometry, and enzyme-linked immunosorbent assay. However, these methods often suffer from time-consuming process, high costs, low sensitivity, and the requirement for specialized personnel. Therefore, a new rapid, sensitive, and specific tumor detection method is urgently needed.The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system, originating from the adaptive immune system of bacteria, has found extensive applications in gene editing and nucleic acid detection. Based on the structure and function of Cas proteins, the CRISPR/Cas system can be classified into two classes and six types. Class I systems consist of multiple Cas protein complexes, including types I, III, and IV, while Class II systems comprise single, multi-domain Cas proteins mediated by RNA, including types II (Cas9), V (Cas12), and VI (Cas13). Class II systems have been widely employed in the fields of biotechnology and nucleic acid diagnostics due to their efficient target binding and programmable RNA specificity. Currently, fluorescence method is the most common signal output technique in CRISPR/Cas-based biosensors. However, this method often requires the integration of signal amplification technologies to enhance sensitivity and involves expensive and complex fluorescence detectors. To enhance the detection performance of CRISPR/Cas-based biosensors, the integration of CRISPR/Cas with some alternative techniques can be considered. The CRISPR/Cas integrated electrochemical sensor (E-CRISPR) possesses advantages such as miniaturization, high sensitivity, high specificity, and fast response speed.E-CRISPR can convert the reactions between biomolecules and detecting components into electrical signals, rendering the detection signals more easily readable and reducing the impact of background values. Therefore,E-CRISPR enhances the accuracy of detection results. E-CRISPR has been applied in various fields, including medical and health, environmental monitoring, and food safety. Furthermore, E-CRISPR holds tremendous potential for advancing the detection levels of tumor markers.Among all types of Cas enzymes, the three most widely applied are Cas9, Cas12, and Cas13, along with their respective subtypes. In this work, we provided a brief overview of the principles and characteristics of Class II CRISPR/Cas single-effector proteins. This paper focused on the various detection technologies based on E-CRISPR technique, including electrochemical impedance spectroscopy, voltammetry, photoelectrochemistry, and electrochemiluminescence. We also emphasized the applications of E-CRISPR in the field of tumor diagnosis, which mainly encompasses the detection of three typical tumor markers (ctDNA, miRNA, and proteins). Finally, we discussed the advantages and limitations of E-CRISPR, current challenges, and future development prospects. In summary, althoughE-CRISPR platform has made significant strides in tumor detection, certain challenges still need to be overcome for their widespread clinical application. Continuous optimization of the E-CRISPR platform holds the promise of achieving more accurate tumor subtyping diagnoses in clinical settings, which would be of significant importance for early patient diagnosis and prognosis assessment. 
		                        		
		                        		
		                        		
		                        	
10.Species-level Microbiota of Biting Midges and Ticks from Poyang Lake
Jian GONG ; Fei Fei WANG ; Qing Yang LIU ; Ji PU ; Zhi Ling DONG ; Hui Si ZHANG ; Zhou Zhen HUANG ; Yuan Yu HUANG ; Ben Ya LI ; Xin Cai YANG ; Meihui Yuan TAO ; Jun Li ZHAO ; Dong JIN ; Yun Li LIU ; Jing YANG ; Shan LU
Biomedical and Environmental Sciences 2024;37(3):266-277,中插1-中插3
		                        		
		                        			
		                        			Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control. Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake. Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors. Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.
		                        		
		                        		
		                        		
		                        	
            
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