OBJECTIVE To observe the effects of endogenous metabolite mesaconate combined with Sodium lactate Ringer’s injection （LR） on prolonging the golden treatment time window and its resuscitation efficacy in rats with hemorrhagic shock under high-altitude conditions. METHODS Rats were divided into the shock group， LR group， and 5， 20， 50 mg/kg mesaconate+LR groups， with 20 rats in each group， to investigate the effect of additional use of mesaconate on the golden treatment time window. After establishing a model of uncontrolled hemorrhagic shock under high-altitude conditions in all groups by housing in a hypobaric hypoxia chamber combined with splenic artery transection， rats in the shock group received no resuscitation， while rats in the LR group and mesaconate+LR groups underwent low-pressure resuscitation with LR or mesaconate combined with LR. Blood pressure control， fluid infusion volume， blood loss rate and survival status were observed in each group. Rats were further divided into the normal group， shock group and mesaconate （50 mg/kg）+LR group， with 10 or 20 rats in each group， to evaluate the resuscitation effects after extending the golden treatment time window by additionally using mesaconate. Except for the normal group， the other groups underwent the same procedure to establish an uncontrolled hemorrhagic shock model under high-altitude conditions. Rats in the shock group received no resuscitation. In the mesaconate+LR group， after 3 h of low-pressure resuscitation， bleeding control was performed by ligation of the spleen artery， and the infusion volume and blood loss rate were recorded； subsequently， the rats received LR resuscitation with twice the volume of blood loss. Then， blood gas indicators of the mesaconate+LR group were measured at different time points. Survival rates， indicators related to sublingual microcirculatory perfusion， liver and kidney blood flow， indicators related to the function of vital organs， and lung and brain water content were observed in all groups. RESULTS LR infusion alone could effectively maintain mean arterial pressure （MAP） within 50-60 mmHg for approximately 1 h. The administration of mesaconate combined with LR during hypotensive resuscitation could maintain MAP within 50-60 mmHg for over 3 h， with significantly reduced fluid infusion volume and blood loss rate in 50 mg/kg mesaconate+LR group， compared to the LR group （ P ＜0.05）. In the LR group， rats maintained low pressure for up to 1 hour with a survival rate of 52.94%， and no rats survived beyond 2 h. In the 5， 20 and 50 mg/kg mesaconate+LR groups， rats maintained low pressure for up to 1 h with a survival rate exceeding 80%； in the 20 and 50 mg/kg mesaconate+LR groups， rats maintained low pressure for up to 3 h with a survival rate exceeding 70%. After complete resuscitation with mesaconate combined with LR， the 72 h survival rate of rats was 43.75%， and significant improvements in blood gas parameters were observed compared to the end of the shock phase （ P ＜0.05）. Compared to the shock group， the mesaconate+LR group showed significant recovery in sublingual microcirculatory indicators， and liver/kidney blood flow after complete resuscitation （ P ＜0.05）， with significant reductions in heart， liver and kidney function-related indicators and lung water content （ P ＜0.05）. CONCLUSIONS Mesaconate combined with LR significantly extends the golden treatment time window for hemorrhagic shock in rats under high-altitude conditions， improves blood gas parameters， sublingual microcirculatory perfusion， and liver/kidney blood flow， mitigates vital organ impairment and pulmonary edema， and increases the survival rate of shocked rats.

Objective To explore the protective effect of L-carnitine on myocardial systolic dysfunction in sepsis and its underlying mechanism.Methods A mouse sepsis model was established by cecal ligation and puncture(CLP).Ten-week-old male SPF-grade C57BL/6 mice(body weight 20～30 g)were randomly divided into 5 groups via random number table:Sham group,Sepsis group,L-carnitine group,L-carnitine+Etomoxir(Eto)group,and Eto group.Echocardiography assessed cardiac function,ELISA measured serum creatine kinase isoenzyme MB(CK-MB)levels,and 72-hour survival rates were recorded to evaluate L-carnitine's effects on cardiac function.Cardiomyocytes were isolated,and a cell microtensiometer was used to detect cardiomyocyte contractile function and calcium transients.Myocardial tissues were collected from each group,and ELISA was used to determine the contents of triglyceride(TG),free fatty acid(FFA),and adenosine triphosphate(ATP).An in vitro sepsis model was constructed by stimulating HL-1 cardiomyocytes with lipopolysaccharide(LPS)for 12 hours,which was divided into 5 groups:control(CTRL)group,LPS group,L-carnitine group,L-carnitine+Eto group,and Eto group.ELISA was used to detect the contents of TG,FFA,and ATP as well as the activity of carnitine palmitoyltransferase 1A(CPT1A)in cardiomyocytes.A cellular energy metabolism analysis system was employed to measure fatty acid oxidation capacity,and Western blot was used to detect the protein expression of CPT1A in cardiomyocytes.BODIPY-FL-C16(green fluorescently labeled palmitic acid)was utilized to detect the distribution of fatty acids in the cytoplasm and mitochondria via immunofluorescence technology,thereby observing the ability of cells to transport fatty acids into mitochondria.Results Compared with the Sham group,cardiac function was significantly impaired in the Sepsis group,as evidenced by decreased ejection fraction and mean arterial pressure(P<0.05),along with elevated levels of the cardiac injury marker CK-MB(P<0.05).Treatment with L-carnitine significantly improved myocardial function,restored blood pressure in septic mice,and increased their survival rate from 12.50%to 81.25%(P<0.05).Compared with the Sham group,the contractile function and calcium transients of acutely isolated single cardiomyocytes were significantly reduced in the Sepsis group(P<0.05),while L-carnitine treatment remarkably restored the contractile function and calcium release capacity of septic cardiomyocytes(P<0.05).Both in vivo and in vitro experiments showed that TG and FFA levels were significantly increased(P<0.05),and ATP levels was significantly decreased(P<0.05)in the Sepsis and LPS groups—effects significantly reversed by L-carnitine treatment.Compared with the CTRL group,the basal oxidation rate and maximum oxidation capacity of fatty acids in cardiomyocytes of the LPS group were significantly reduced(P<0.05),and L-carnitine treatment notably improved these indicators.Compared with the CTRL group,the expression and activity of CPT1A in cardiomyocytes of the LPS group were significantly decreased(P<0.05),while L-carnitine treatment significantly increased the expression and activity of CPT1A(P<0.05).In LPS group cardiomyocytes,green fluorescently labeled palmitic acid primarily formed numerous granular/clumpy aggregates in the cytoplasm with minimal mitochondrial colocalization.In the L-carnitine group,the green fluorescent granules in the cytoplasm of cardiomyocytes were smaller,and colocalization with mitochondria was increased.However,the L-carnitine+Eto group exhibited similar phenomena to the LPS group.In addition,both in vivo and in vitro experiments demonstrated that treatment with the CPT1A inhibitor Eto reversed the effect of L-carnitine.Compared with the L-carnitine group,the ATP content in the L-carnitine+Eto group was significantly decreased(P<0.05),while the FFA content was significantly increased(P<0.05).Conclusion L-carnitine facilitates fatty acid entry into mitochondria for β-oxidation via a CPT1A-dependent mechanism,thereby ameliorating fatty acid oxidation dysfunction in septic cardiomyocytes and improving myocardial contractile function.

Objective To investigate the clearance capacity of cardiac resident macrophages in post-sepsis and its underlying mechanism.Methods A mouse model of sepsis was established using cecum perforation ligation.Thirty male C57BL/6 mice(8 weeks old,weighing 20～25 g)were randomly and equally divided into a sham operation group(sham group)and a model group(sepsis group).Immunofluorescence assay was employed to label the cardiomyocytes and macrophages to observe the apoptosis of cardiomyocytes and the phagocytosis by cardiac resident macrophages.Cardiac resident macrophages were extracted for transcriptomic sequencing to determine the functional changes of the cells after sepsis.Cardiac resident macrophage cell lines were established at the cellular level and served as the normal group(RAC group),and the RAC cells treated with LPS were subjected as the sepsis group(RAC+LPS group).Then the differences in the ability to clear apoptotic cardiomyocytes between the 2 groups were observed.Then DQ-BSA-RED lysosomal activity detection probe,Lyso-Sensor yellow/bule dye,ELISA,and Western blotting were applied to detect the lysosomal function of cardiac resident macrophages,activity and expression of important lysosomal hydrolases,changes in contents and related subunits of vacuolar-type adenosine triphosphatases(V-ATPase).Results Compared with the sham group,the sepsis group had larger number of apoptotic cardiomyocytes(P＜0.05)and increased phagocytosis of cardiomyocytes by cardiac macrophages(P＜0.05).The results of transcriptomic sequencing revealed a significant dysfunction of lysosome-associated functions of cardiac-resident macrophages after sepsis.In in vitro experiments,the RAC+LPS group had a reduced fragmentation capacity of apoptotic cardiomyocytes,reduction in the intensity of yellow fluorescence of lysosomes(P＜0.05),and decrease in lysosomal hydrolase activity(P＜0.05)when compared with the RAC group.In addition,LPS treatment significantly decreased the activity and expression of V-ATPase and its major subunit ATP6V1A in cardiac resident macrophages(P＜0.05).Conclusion Cardiac resident macrophages show reduced clearance of apoptotic cardiomyocytes after sepsis,which may be related to a decrease in the activity of ATP6V1A,an important subunit of its lysosomal V-ATPase,and reduced activity of lysosomal hydrolases.

Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

Aristolochic acids (AAs) have long been considered as a potent carcinogen due to its nephrotoxicity. Aristolochic acid I (AAI) reacts with DNA to form covalent aristolactam (AL)-DNA adducts, leading to subsequent A to T transversion mutation, commonly referred as AA mutational signature. Previous research inferred that AAs were widely implicated in liver cancer throughout Asia. In this study, we explored whether AAs exposure was the main cause of liver cancer in the context of HBV infection in mainland China. Totally 1256 liver cancer samples were randomly retrieved from 3 medical centers and a refined bioanalytical method was used to detect AAI-DNA adducts. 5.10% of these samples could be identified as AAI positive exposure. Whole genome sequencing suggested 8.41% of 107 liver cancer patients exhibited the dominant AA mutational signature, indicating a relatively low overall AAI exposure rate. In animal models, long-term administration of AAI barely increased liver tumorigenesis in adult mice, opposite from its tumor-inducing role when subjected to infant mice. Furthermore, AAI induced dose-dependent accumulation of AA-DNA adduct in target organs in adult mice, with the most detected in kidney instead of liver. Taken together, our data indicate that AA exposure was not the major threat of liver cancer in adulthood.

Objective:To investigate the feasibility and efficacy of three mini-incision double eyelid blepharoplasty with concurrent reposition of orbital fat pad to correct sunken upper eyelids.Methods:The data of patients who underwent orbital fat reposition to correct upper eyelid depression during the period from January 2018 to April 2021 in the Medical Plastic and Cosmetic Center of Beijing Tongren Hospital affiliated to Capital Medical University were retrospectively analyzed. During the operation, the central mass of orbital fat and its lateral extension were fully dissociated, and then transferred and fixed inward through three mini-incisions. Postoperative follow-up was performed to observe the improvement of the double eyelid shape and upper eyelid depression, and the patient satisfaction was recorded.Results:A total of 72 patients (140 eyes) were included in this study, including 1 male and 71 females, with an average age of 28 years (19-40 years old), 122 eyes with mild upper eyelid depression and 18 eyes with moderate upper eyelid depression before operation. Postoperative follow-up was 6-24 months, with an average of 11 months. The curvature of the double eyelid after operation was smooth and basically symmetrical. Among the 122 eyes with mild upper eyelid depression before operation, 118 eyes were completely corrected after operation, and 4 eyes were not improved. Of the 18 eyes, 3 eyes were completely corrected after operation, and 15 eyes still had mild upper lid depression. The overall satisfaction rate was 93.1% (67/72), the rate of double eyelid failure was 3.6% (5/140), and no other serious complications were found.Conclusions:Three mini-incision double eyelid blepharoplasty simultaneously with orbital fat reposition can effectively correct mild and moderate sunken upper eyelid deformity. The postoperative double eyelid shape is stable with high satisfaction rate.

Objective:To investigate the feasibility and efficacy of three mini-incision double eyelid blepharoplasty with concurrent reposition of orbital fat pad to correct sunken upper eyelids.Methods:The data of patients who underwent orbital fat reposition to correct upper eyelid depression during the period from January 2018 to April 2021 in the Medical Plastic and Cosmetic Center of Beijing Tongren Hospital affiliated to Capital Medical University were retrospectively analyzed. During the operation, the central mass of orbital fat and its lateral extension were fully dissociated, and then transferred and fixed inward through three mini-incisions. Postoperative follow-up was performed to observe the improvement of the double eyelid shape and upper eyelid depression, and the patient satisfaction was recorded.Results:A total of 72 patients (140 eyes) were included in this study, including 1 male and 71 females, with an average age of 28 years (19-40 years old), 122 eyes with mild upper eyelid depression and 18 eyes with moderate upper eyelid depression before operation. Postoperative follow-up was 6-24 months, with an average of 11 months. The curvature of the double eyelid after operation was smooth and basically symmetrical. Among the 122 eyes with mild upper eyelid depression before operation, 118 eyes were completely corrected after operation, and 4 eyes were not improved. Of the 18 eyes, 3 eyes were completely corrected after operation, and 15 eyes still had mild upper lid depression. The overall satisfaction rate was 93.1% (67/72), the rate of double eyelid failure was 3.6% (5/140), and no other serious complications were found.Conclusions:Three mini-incision double eyelid blepharoplasty simultaneously with orbital fat reposition can effectively correct mild and moderate sunken upper eyelid deformity. The postoperative double eyelid shape is stable with high satisfaction rate.

Objective To analyze the drug consumption of the joint logistics support force during field training in Yunnan, summarize the drug use characteristics of medical staff, and analyze the drug use rules, so as to provide the basis for modular drug support. Methods The basic information and drug use of the personnel in an institution of the joint logistics support force during the resident training period from April to June 2020 were collected and sorted out. The drug use indicators were calculated by using the defined daily dose (DDD) as the unit, including: daily drug dose (DDDs), daily drug dose cost (DDDc), drug use intensity (DUI), drug utilization rate (DUR) and drug ranking ratio (R) were used to describe drug use characteristics. Results During the resident training period, the amount of musculoskeletal system drugs was large, and the number of users was large. The dosage of cardiovascular system drugs, digestive system drugs and hormone drugs was large, but the number of users was small. Sensory organ drugs, dermatological drugs, heat-clearing agents (Chinese patent medicine) and respiratory system drugs were small in dosages, but the number of users was large. Ranked by DDDs, the top 3 drugs with the highest dosage were Yunnan Baiyao plaster, Yunnan Baiyao Band-Aid and Loratadine Tablets. Ranked by DDDc, the largest daily cost was Budesonide and Formoterol Fumarate Powder for Inhalation. As age increases, the intensity of drug use increased; The intensity of drug use in women was greater than that in men. The intensity of drug use was different for different categories personnel. Conclusion Musculoskeletal system drugs are important drugs for field training. The task personnel should formulate a list of medicines for basic diseases based on their own medication for basic diseases and the duration of the task, and submit it to the medicinal material guarantee department to ensure the carrying drugs. The factors affecting modular drug support in field training include: task type, region, solar term, duration and personnel composition, etc.

Objective To investigate the early resuscitation effect of hemoglobin-based oxygen carriers (HBOC) in rats with uncontrolled hemorrhagic shock.Methods 170 Sprague-Dawley (SD) rats were randomly divided into five groups:lactate Ringer solution (LR) control group,whole blood control group,and 0.5％,2.0％,5.0％ HBOC groups,with 34 rats in each group.The uncontrolled hemorrhagic shock model in SD rats was reproduced by cutting off the splenic artery branch,and induced mean arterial pressure (MAP) reducing to 40 mmHg (1 mmHg =0.133 kPa).The corresponding solution was infused after model reproduction in each group,maintaining MAP at 50 mmHg for 1 hour,then completely ligating and hemostasis,and maintaining MAP at 70 mmHg for 1 hour and 80 mmHg for 1 hour respectively,after maintaining MAP 80 mmHg,all were supplemented with LR to 2 times blood loss volume.The survival rate and blood loss rate were observed in 16 rats in each group,hemodynamics parameters including MAP,left ventricular systolic pressure (LVSP) and the maximum rate of left ventricular pressure rise (+dp/dtmax) were determined in another 10 rats,and cardiac output (CO) and tissue oxygen supply (DO2) were observed in the rest 8 rats.Results ① When resuscitation by LR alone,the blood loss rate of animals was as high as 60％ to 70％.Compared with the LR control group,whole blood recovery could significantly reduce the blood loss rate before hemostasis in uncontrolled hemorrhagic shock rats [(46.6 ± 4.5)％ vs.(62.3 ± 4.0)％,P ＜ 0.01];0.5％,2.0％,5.0％ HBOC could significantly decrease the blood loss rate,especially in 5.0％ HBOC group with significant difference as compared with that in the LR control group [(45.6±4.1)％ vs.(62.3±4.0)％,P ＜ 0.01].② When LR was used alone for resuscitation,the rats died quickly and survived for a short time.Only one rat survived for 12 hours,and no rat survived for more than 24 hours.Compared with the LR control group,whole blood resuscitation could improve the survival rate of uncontrolled hemorrhagic shock rats,and the survival time was significantly prolonged (hours:20.4± 4.6 vs.3.5 ± 1.1,P ＜ 0.01);0.5％,2.0％ and 5.0％ HBOC also significantly prolonged the survival time of rats.The 5.0％ HBOC group had the best effect,4 rats survived in 24 hours,and the survival time was significantly longer than that of the LR control group (hours:18.4 ± 4.0 vs.3.5 ± 1.1,P ＜ 0.01),and it was the same as the whole blood control group.③ Compared with pre-shock,CO,DO2 and hemodynamic parameters of uncontrolled hemorrhagic shock rats were significantly decreased,and the above parameters were gradually increased with the prolongation of rehydration time.Compared with the LR control group,whole blood resuscitation could significantly increase CO and DO2,and improve hemodynamics in rats with uncontrolled hemorrhagic shock at different time points.Three concentrations of HBOC could also increase CO,DO2 and other hemodynamic parameters of rats at 1 hour of maintaining MAP of 80 mmHg after hemostasis and 1 hour and 2 hours after resuscitation.The effect of 5.0％ HBOC group was more significant than that of the LR control group with statistically significant difference [CO (× 10-3,L/min):72.84±2.84 vs.63.11±2.38 at 1 hour of maintaining MAP of 80 mmHg,70.25±4.55 vs.59.88 ± 9.31 at 1 hour after resuscitation,71.51 ± 2.90 vs.53.24 ± 6.32 at 2 hours after resuscitation;DO2 (L· min-1 · m-2):271.9± 13.5 vs.159.1 ±25.4 at 1 hour of maintaining MAP of 80 mmHg,261.0± 15.0 vs.145.7±20.1 at 1 hour after resuscitation,249.6± 12.0 vs.107.4± 18.2 at 2 hours after resuscitation;MAP (mmHg):82.1±2.1 vs.74.0±2.8 at 1 hour of maintaining MAP of 80 mmHg,107.5±9.3 vs.64.0±5.7 at 1 hour after resuscitation,104.0±9.7 vs.73.0±4.2 at 2 hours after resuscitation;LVSP (mmHg):128.6±7.9 vs.103.8±0.8 at 1 hour of maintaining MAP of 80 mmHg,129.3±± 15.0 vs.99.4±0.0 at 1 hour after resuscitation,127.5± 11.3 vs.97.4±0.0 at 2 hours after resuscitation;+dp/dt max (mmHg/s):6 534.2±± 787.6 vs.5 074.0± 71.7 at 1 hour of maintaining MAP of 80 mmHg,5 961.5 ±± 545.4 vs.4 934.5 ± 510.2 at 1 hour after resuscitation,5 897.4± 350.5 vs.4 534.7 ±489.2 at 2 hours after resuscitation,all P ＜ 0.05].Conclusions HBOC infusion prolonged the survival time,increased survival rate,and improved hemodynamics,cardiac function and tissue oxygen supply in a dose-dependent manner in the early stage of uncontrolled hemorrhagic shock.The recovery effect of 5.0％ HBOC was similar to that of the whole blood.

Objective To observe the effectiveness of transplantation of neuron-like cells induced by GDNF from BMSCs on treatment of spinal cord injury in rats.Methods Transfected BMSCs by recombinant adenovirus containing the gene of rGDNF and GFP.36 SD rats with spinal cord injury were randomly divided into three groups.All the 36 SD rats with spinal cord injury were scored by the Basso,Beattle,Bresnahan locomotoRrating scale at different time points (1、2、3、4 weeks afteRoperation).In the 4th week afteRoperation,tissue from the injury section was prepared to observe repaiRof spinal cord injury.Results BMSCs was able to secrete GDNF continuously afteRtransfected by recombinant adenovirus containing the gene of rGDNF and GFP.AfteRoperation,BBB scores in intralesional treatment group significantly increased when compared with that of intravenous treatment group and injury control group.In the 4th week afteRoperation,HEmatoxylin-eosin staing results showed that cavitation in intravenous treatment group was largeRthan that in intralesional treatment group with the one of injury control group being the biggest.Immunohistochemical method showed that there were more NF200+cells and GFAP+cells in intralesional treatment group than in the otheRtwo groups,and injury control group had the least NF200+cells and GFAP+ cells.Conclusion AfteRinduced into nuron-like cells,BMSCs can treat spinal cord injury effectively.