(+)-Borneol, the main component of "Natural Borneol" in the Chinese Pharmacopoeia, is a high-end spice and precious medicine. Plant extraction cannot meet the increasing demand for (+)-borneol, while microbial biosynthesis offers a sustainable supply route. However, its production was extremely low compared with other monoterpenes, even with extensively optimizing the mevalonate pathway. We found that the key challenge is the complex and unusual dephosphorylation reaction of bornyl diphosphate (BPP), which suffers the side-reaction and the competition from the cellular dephosphorylation process, especially lipid metabolism, thus limiting (+)-borneol synthesis. Here, we systematically optimized the dephosphorylation process by identifying, characterizing phosphatases, and balancing cellular dephosphorylation metabolism. For the first time, we identified two endogenous phosphatases and seven heterologous phosphatases, which significantly increased (+)-borneol production by up to 152%. By engineering BPP dephosphorylation and optimizing the MVA pathway, the production of (+)-borneol was increased by 33.8-fold, which enabled the production of 753 mg/L under fed-batch fermentation in shake flasks, so far the highest reported in the literature. This study showed that rewiring dephosphorylation metabolism was essential for high-level production of (+)-borneol in Saccharomyces cerevisiae, and balancing cellular dephosphorylation is also helpful for efficient biosynthesis of other terpenoids since all whose biosynthesis involves the dephosphorylation procedure.

Objective To investigate the effect of whey protein supplementation on rehabilitation of stroke-related sarcopenia patients with diabetes mellitus.Methods Forty-seven stroke-related sarcopenia patients with diabetes mellitus admitted to the First Affiliated Hospital of Wenzhou Medical University from January 2021 to September 2023 were selected and divided into observation group(24 cases)and control group(23 cases)according to random number table method.Patients in both groups were given normal diabetic diet,conventional drug therapy and rehabilitation training,and patients in observation group were given whey protein powder orally every day for 4 weeks.The non-hemiplegic hand grip strength,appendicular skeletal muscle mass index(ASMI),modified Barthel index(MBI),Fugl-Meyer assessment(FMA),mini-nutritional assessment short-form(MNA-SF),serum albumin(Alb),C-reactive protein(CRP)were compared between two groups before and after intervention.Results After intervention,the grip strength,ASMI,FMA,MBI of two groups were significantly higher than before ntervention,and CRP level was significantly lower than before intervention(P<0.05).The grip strength,ASMI,FMA,MBI,MNA-SF score and Alb of observation group were significantly higher than those of control group,while CRP was significantly lower than that of control group(P<0.05).ASMI was positively correlated with Alb,MNA-SF score and grip strength(P<0.05),while CRP was negatively correlated with Alb,MNA-SF score and ASMI(P<0.05).Conclusion Supplementation of whey protein can improve the nutritional status,muscle strength and muscle mass of patients with stroke-related sarcopenia complicated with diabetes mellitus,and improve the physical motor function and the ability of daily living.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To establish a nomogram model for predicting the hyper-progression recurrence after hepatectomy in patients with hepatocellular carcinoma (HCC) based on circulating tumor cells (CTC).Methods:Clinical data of 231 HCC patients undergoing hepatectomy at the Department of Hepatobiliary Surgery, Guangxi Medical University Cancer Hospital from January 2013 to December 2022 were retrospectively analyzed, including 200 males and 31 females, aged 46(39, 52) years old. Patients were divided into two groups: the modeling group ( n=154) and the validation group ( n=77). According to the state of postoperative hyper-progression recurrence, patients in the modeling group were subdivided into hyper-progression recurrence ( n=39) and non-hyper-progression recurrence group ( n=115). Patients in the validation group were also subdivided into hyper-progression recurrence ( n=16) and non-hyper-progression recurrence group ( n=61). Clinicopathological data such as the total CTC count, alpha-fetoprotein, and postoperative pathology were collected. Logistic regression analysis was used to analyze the influencing factors of postoperative hyper-progression recurrence. A nomogram model was established based on the results of multivariate logistic regression analysis. The receiver operating characteristic (ROC) curve, calibration curve, decision curve analysis (DCA) and clinical impact curve (CIC) were used to validate the nomogram model. Results:Multivariate logistic regression analysis showed that HCC patients with age ≤45 years old ( OR=6.704, 95% CI: 1.619-27.760, P=0.009), incomplete tumor capsule ( OR=13.292, 95% CI: 3.084-57.295, P=0.001), high total numbers of CTC ( OR=1.101, 95% CI: 1.023-1.186, P=0.011) and high Ki67 index ( OR=52.659, 95% CI: 3.215-862.604, P=0.005) had a high risk of hyper-progression recurrence after hepatectomy. The above three preoperative variables were integrated to construct a nomogram model. The calibration curve showed that the predicted results of the nomogram model were in good agreement with the actual results. The ROC curves of the nomogram model for predicting hyper-progression recurrence after hepatectomy in HCC patients were plotted, and the area under the curve was 0.907 (95% CI: 0.856-0.959) and 0.833 (95% CI: 0.721-0.945) in the modeling group and validation group, respectively. DCA showed that the nomogram model could be used as a valuable predictive tool for the hyper-progression recurrence after hepatectomy. The CIC showed that the population judged by the nomogram model was highly matched with the actual population with hyper-progression recurrence. Conclusions:This study established a nomogram model based on age, tumor capsular integrity and total CTC count, which could accurately predict the postoperative hyper-progression recurrence in HCC patients before hepatectomy. The model is promising in guiding clinical practice after further validation.

Objective To explore the trajectory and influencing factors of joint awareness after total knee arthroplasty.Methods With the method for convenience sampling,patients who met the standards for total knee arthroplasty from July 2021 to March 2022 were selected.General information,severity of osteoarthritis,anxiety and depression,and self-efficacy were investigated before surgery(T0).Joint awareness was investigated at 1 month(T1),3 months(T2),and 6 months(T3)after surgery,respectively.Latent growth curve model was used to describe the overall trend of joint awareness;latent class growth model was used to analyze latent subgroups;logistic regression analysis was used to determine the impact of related variables on the trajectory of joint awareness.Results The scores of joint awareness were(32.70±5.80),(47.67±4.67)and(61.53±7.81)respectively at T1 to T3,and joint awareness showed an increasing trend(P＜0.001).The trajectory of joint awareness was divided into 2 potential subgroups:the rapid growth group(61.83％)and the slow growth group(38.17％).Age,BMI,other chronic diseases or not,years of osteoarthritis,severity of osteoarthritis,anxiety,depression,and self-efficacy affected the trend of joint awareness(all P＜0.05).Conclusion Joint awareness showed a linear growth trend with the postoperative time,and there were 2 potential subgroups in the trajectory.Medical workers could develop corresponding interventions based on the influencing factors to improve patients'postoperative awareness of artificial joints.

Objective:To investigate the relationship between dyslipidemia and the risk of non-alcoholic fatty liver disease (NAFLD) in adults.Methods:A total of 13 449 individuals with an average age of 37 (31,48) years including 4 815(35.80%) males, who underwent physical examination and had no NAFLD in Tianjin Medical University General Hospital in 2017 were recruited in this prospective study. Participants were followed up till December 31, 2021, and the newly onset NAFLD was defined as endpoint of the study. Cox regression model was used to analyze the correlation between the dyslipidemia and the risk of NAFLD.Results:The mean follow-up period was 2.80 years with total 37 707 person-years of follow-up. Among 13 449 participants, 2 711 subjects developed NAFLD during the follow-up period with a cumulative incidence rate of 20.2%. After adjusting for confounding factors, compared with the non-dyslipidemia group, the hazard ratio (HR) (95% CI) of the 1, 2, and 3 types of dyslipidemia groups were 1.388 (1.253-1.538), 1.200 (1.143-1.261) and 1.282 (1.226-1.342), respectively (trend test: P<0.001); and the HR (95% CI) of the hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia and low high-density lipoprotein cholesterolemia groups were 1.165 (1.058-1.283), 1.436 (1.345-1.533), 1.291 (1.240-1.344) and 1.068 (1.014-1.124) respectively (all P<0.05). After adjusting for confounding factors, compared with the normal triglycerides group, HR (95% CI) of moderately elevated triglycerides and elevated triglycerides group were 1.774 (1.590-1.980) and 1.443 (1.345-1.549) respectively (trend test: P<0.001). Conclusion:Dyslipidemia can increase the risk of NAFLD, especially for individuals with hypertriglyceridemia, which indicates that appropriate management of blood lipid levels, especially triglyceride level may effectively prevent NAFLD.

Objective To assess the impact of intravenous esketamine administered prior to car-diopulmonary bypass(CPB)initiation on ventricular function and internal carotid artery blood flow in pa-tients undergoing heart valve replacement surgery.Methods Sixty patients underwent elective CPB heart valve replacement,38 males and 22 females,aged 18-75 years,BMI 18.5-30.0 kg/m2,ASA physical status Ⅱ or Ⅲ,NYHA cardiac function classification Ⅰ-Ⅲ,and a left ventricular ejection fraction(LVEF)of≥45％,were selected.The patients were randomly divided into two groups:esketamine group(group E)and normal saline group(group C),30 patients in each group.Total intravenous anesthesia was used during the operation.Following the initiation of CPB,group E received an intravenous infusion of es-ketamine at a rate of 0.5 mg·kg-1·h-1 until the conclusion of the procedure,while group C received an equivalent volume of normal saline concurrently at the same rate.HR,MAP,CVP,and cardiac output index(CI)were recorded before anesthesia induction,during skin resection,and within 60 minutes after stopping CPB.LVEF,left ventricular global longitudinal strain(GLS),global longitudinal time-to-peak strain standard deviation(GLTSD),global circumferential strain(GCS),global circumferential time-to-peak strain standard deviation(GCTSD),right ventricular ejection fraction(RVEF),right ventricular GLS,and GLTSD were obtained during skin resection,within 40 minutes of CPB,and 60 minutes after stopping CPB.rScO2,BIS,concentrations of Hb and lactic acid(Lac),peak systolic flow velocity(SPV),quantity of flow-internal carotid artery(Q-ICA),and blood flow resistance index(RI)were recorded before anesthesia induction,during skin resection,within 40 minutes of CPB,and within 60 minutes after stopping CPB.Concentrations of cardiac troponin Ⅰ(cTnⅠ),alanine aminotransferase(ALT),creatinine(Cr),and neuron-specific enolase(NSE)were recorded before anesthesia induction and 6 hours after operation.Spon-taneous resuscitation after CPB,postoperative extubation time,duration of ICU stay,total hospital stay,in-cidence of adverse cardiac events,and 30-day postoperative mortality were recorded.Results Compared with group C,group E exhibited a significant increase in CI within 60 minutes after stopping CPB(P＜0.05).The LVEF,RVEF,and right ventricular GLS demonstrated significant increases within 60 minutes after stopping CPB in group E compared with group C(P＜0.05).The left ventricular GLS and left ven-tricular GCTSD displayed significant increases 30 minutes after stopping CPB in group E compared with group C.The RI exhibited a significant increase within 40 minutes of CPB in group E compared with group C(P＜0.05).There were no significant differences in cTnⅠ,ALT,Cr,NSE,spontaneous resuscitation affter CPB,postoperative extubation time,duration of ICU stay,total hospital stay,incidence of cardiac adverse events,and 30-day postoperative mortality between the two groups.Conclusion Administration of esket-amine following the onset of CPB in patients undergoing cardiac surgery demonstrates a significant elevation in CI post-CPB cessation.Furthermore,it may augment ventricular longitudinal strain,thereby enhancing myocardial contraction,leading to increased postoperative ventricular ejection fraction,and sustaining hemo-dynamic stability.

Objective:To explore the clinical features and genetic variants in three children suspected for β-ketothiolase deficiency (BKTD).Methods:Clinical manifestations, laboratory examination and genetic testing of three children suspected for BKTD at Henan Children′s Hospital between January 2018 and October 2022 were collected, and their clinical and genetic variants were retrospectively analyzed.Results:The children were all males with a age from 7 to 11 months. Their clinical manifestations have included poor spirit, shortness of breath, vomiting, convulsions after traumatic stress and/or infection. All of them had severe metabolic acidosis, elevated ketone bodies in blood and urine, hypoglycemia, with increased isoprenyl-carnitine and 3-hydroxyisovalyl-carnitine in the blood, and 2-methyl-3-hydroxybutyrate and methylprotaroyl glycine in the urine. All of them were found to harbor compound heterozygous variants of the ACAT1 gene, including c. 1183G>T and a large fragment deletion (11q22.3-11q23.1) in child 1, c. 121-3C>G and c. 826+ 5_826+ 9delGTGTT in child 2, and c. 928G>C and c. 1142T>C in child 3. The variants harbored by children 2 and 3 were known to be pathogenic or likely pathogenic. The heterozygous c. 1183G>T variant in child 1 was unreported previously and rated as a variant of unknown significance (PM2_Supporting+ PP3+ PP4) based on guidelines from the American College of Medical Genetics and Genomics. The large segment deletion in 11q22.3-11q23.1 has not been included in the DGV Database and was rated as a pathogenic copy number variation. Conclusion:The variants of the ACAT1 gene probably underlay the pathogenesis of BKTD in these three children.

Background::Ainuovirine (ANV) is a new generation of non-nucleoside reverse transcriptase inhibitor for the treatment of human immunodeficiency virus (HIV) type 1 infection. This study aimed to evaluate the population pharmacokinetic (PopPK) profile and exposure-response relationship of ANV among people living with HIV.Methods::Plasma concentration-time data from phase 1 and phase 3 clinical trials of ANV were pooled for developing the PopPK model. Exposure estimates obtained from the final model were used in exposure-response analysis for virologic responses and safety responses.Results::ANV exhibited a nonlinear pharmacokinetic profile, which was best described by a two-compartment model with first-order elimination. There were no significant covariates correlated to the pharmacokinetic parameters of ANV. The PopPK parameter estimate (relative standard error [%]) for clearance adjusted for bioavailability (CL/F) was 6.46 (15.00) L/h, and the clearance of ANV increased after multiple doses. The exposure-response model revealed no significant correlation between the virologic response (HIV-RNA <50 copies/mL) at 48 weeks and the exposure, but the incidence of adverse events increased with the increasing exposure ( P value of steady-state trough concentration and area under the steady-state curve were 0.0177 and 0.0141, respectively). Conclusions::Our PopPK model supported ANV 150 mg once daily as the recommended dose for people living with HIV, requiring no dose adjustment for the studied factors. Optimization of ANV dose may be warranted in clinical practice due to an increasing trend in adverse reactions with increasing exposure.Trial registration::Chinese Clinical Trial Registry https://www.chictr.org.cn (Nos. ChiCTR1800018022 and ChiCTR1800019041).

Objective To determine the effects of treadmill training on the structure of hippocampal myelin and cognitive function in rats exposed to acute plateau hypoxia.Methods With 30 SPF-grade female SD rats (aged 6-8 weeks,weighing 200-220 g),6 of them were used for observation of myelin structure after injury,and the remaining 24 rats were randomly divided into control group,hypobaric hypoxia group and treadmill training group (n=8).The rats in above experimental groups were placed in a low-pressure oxygen chamber at an altitude of 6000 m for 7 consecutive days,and the rats of the control group were placed in the confined chamber for the same period without hypoxia.Then,the rats of the treadmill training group received a 4-week treadmill training scheme since the day after hypoxia.Finally,all the rats were tested for cognitive function with open field test (OFT)and Morris water maze (MWM).Transmission electron microscopy (TEM) was used to observe the changes of demyelination in the hippocampus. The expression of oligodendrocyte transcription factor 2 (Olig2)and myelin basic protein (MBP )in the hippocampal CA1 and CA3 regions was measured by immunofluorescence staining and Western blotting.Results Behavioral tests showed that the number into the central area,total distance,distance ratio in OFT and the number of platform crossings and distance to the target area in MWM were reduced in the hypobaric hypoxia group than the control group (P<0.05 ),while these indexes were increased in the treadmill training group than in the hypobaric hypoxia group (P<0.05).Immunofluorescence staining indicated that the number of Olig2 positive cells per unit area and the mean fluorescence intensity of MBP in the CA1 and CA3 regions were significantly lessen in the hypobaric hypoxia group than the control group (P<0.05 ),while these indicators were higher in the treadmill training group than the hypobaric hypoxia group (P<0.05 ).Western blotting displayed that the expression levels of Olig2 and MBP in the hippocampus were obviously lower in the hypobaric hypoxia group than the control group (P<0.01 ),while the levels were increased in the treadmill training group than the hypobaric hypoxia group (P<0.01 ).Conclusion Treadmill training promotes the number of the oligodendrocyte spectrum cells in CA1 and CA3 regions,enhances the expression of myelin-related proteins and improves myelin repair in hippocampus of hypobaric hypoxia rats,and thereby ameliorates hypoxia-induced anxiety-like behaviors and memory dysfunction.