(+)-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 prevalence of scoliosis and congenital heart disease(CHD)and their correlation among children and adolescents of Drung nationality in Yunnan Province.Methods A cross-sectional survey was conducted in November 2022 among all Drung school-aged children and adolescents aged 5-18 years in Gongshan Drung and Nu Autonomous County,Yunnan Province.Visual inspection,Adams for-ward flexion test,and trunk rotation angle(ATR)measurement were comprehensively used for school prelim-inary screening of scoliosis.Individuals who tested positive in the school preliminary screening underwent fur-ther X-ray examination for auxiliary diagnosis.Cardiac auscultation and echocardiography were used for school preliminary screening of CHD.The personal information of the screening subjects,the screening results,etc.were recorded.The prevalence of scoliosis and CHD among children and adolescents of the Drung nationality and the relationship between the two diseases were statistically analyzed,and the positive predictive value of school-based scoliosis screening and its influencing factors were also analyzed.Results A total of 1 036 chil-dren and adolescents of Drung nationality were enrolled,with a mean age of(10.72±3.75)years,icluding 542 males and 494 females.A total of 45 subjects tested positive for scoliosis in the school preliminary screening,with a preliminary positive rate of 4.34%.A total of 22 cases were finally diagnosed with scoliosis,with a prevalence rate of 2.12%.Among them,21 cases were idiopathic scoliosis(accounting for 95.45%),and 1 case was congenital scoliosis(accounting for 4.55%).The prevalence rate was higher in females(2.83%)than that in males(1.48%),higher in the 10 to 18-year-old group(2.30%)than that in the 5 to<10-year-old group(1.87%),and higher in the secondary school group(2.78%)than that in the primary school group(1.78%),hut there were no statistically significant differences(P>0.05).Most idiopathic scoliosis cases were mild(Cobb angle 10° to<20°,90.48%)and classified as Lenke type Ⅴ(57.14%).Two cases of CHD were confirmed,both of which were atrial septal defects,with a prevalence rate of 0.19%.The co-occurrence rate of idiopathic scoliosis and CHD was 4.76%(1/21).The positive predictive value of school-based scoliosis pre-liminary screening was only 48.89%.When the BMI was<18.5 kg/m2,the positive predictive value was sig-nificantly higher than that for BMI≥18.5 kg/m2(P<0.05).Conclusion The prevalence rate of scoliosis a-mong adolescents of the Drung ethnic group in Yunnan Province is 2.12%,predominantly idiopathic scoliosis,with Lenke type V being the most common classification.The prevalence rate of congenital heart disease is 0.19%.BMI is a significant influencing factor for the positive predictive value of school-based scoliosis prelimi-nary screening.

Canine mammary tumors are one of the most common causes of cancer-related death in female dogs,challenging traditional surgical therapies due to their high recurrence and metastasis.The natural compound Ole is strongly associated with a variety of phenomena,such as increased longevity,reduced disease incidence and mortality.However,its effect on canine mammary tumors is unclear.Therefore,the following experiments were conducted to investigate whether Ole has an effect on the development of canine mammary tumors:The effects of Ole at different concentra-tions on CHMm cell viability,migration ability,apoptosis,expression levels of corresponding pro-teins Bax and BCL-2 and mRNA expression of tumor-related genes were detected by scratch test,flow cytometry,RT-qPCR and protein western blot,and the mechanism of action was also ex-plored.The results showed that Ole activated the PI3K/AKT signaling pathway,induced phospho-rylation of related proteins,and significantly inhibited the expression levels of p-PI3K and p-AKT after applying OLE to CHMm of canine mammary tumor cells.At the same time,Ole significantly inhibited the survival,migration and proliferation of canine mammary tumor cells.In addition,the expressions of proteins Bax and BCL-2,which are closely related to apoptosis,and mRNA expres-sions of some tumor-related genes were significantly regulated,suggesting that Ole mediated PI3K/AKT signaling pathway effectively induced apoptosis of canine mammary tumor cells.This study suggests that Ole has the potential to become a key drug in the treatment of mammary canc-er,and provides a key target for the development of effective therapies against canine mammary tumors.

Canine mammary tumors are one of the most common causes of cancer-related death in female dogs,challenging traditional surgical therapies due to their high recurrence and metastasis.The natural compound Ole is strongly associated with a variety of phenomena,such as increased longevity,reduced disease incidence and mortality.However,its effect on canine mammary tumors is unclear.Therefore,the following experiments were conducted to investigate whether Ole has an effect on the development of canine mammary tumors:The effects of Ole at different concentra-tions on CHMm cell viability,migration ability,apoptosis,expression levels of corresponding pro-teins Bax and BCL-2 and mRNA expression of tumor-related genes were detected by scratch test,flow cytometry,RT-qPCR and protein western blot,and the mechanism of action was also ex-plored.The results showed that Ole activated the PI3K/AKT signaling pathway,induced phospho-rylation of related proteins,and significantly inhibited the expression levels of p-PI3K and p-AKT after applying OLE to CHMm of canine mammary tumor cells.At the same time,Ole significantly inhibited the survival,migration and proliferation of canine mammary tumor cells.In addition,the expressions of proteins Bax and BCL-2,which are closely related to apoptosis,and mRNA expres-sions of some tumor-related genes were significantly regulated,suggesting that Ole mediated PI3K/AKT signaling pathway effectively induced apoptosis of canine mammary tumor cells.This study suggests that Ole has the potential to become a key drug in the treatment of mammary canc-er,and provides a key target for the development of effective therapies against canine mammary tumors.

low-grade glioma(LGG)is a slow-growing primary brain tumor originating from glial cells.The overall prognosis of LGG is good but incurable.The vast majority of LGG will have relapses,and some even progress to high-grade glioma(HGG).The treatment of LGG is still controversial.The current treatment includes surgical resection,radiation therapy(RT),chemical treatment(CT),targeting and immunotherapy,but its effectiveness and safety need further clinical exploration.This article describes the treatment of LGG,including surgery,radiotherapy,chemotherapy,targeting and immunotherapy,in order to develop an individualized treatment plan and provide theoretical strategies for maximizing the survival time of patients and improving the prognosis of patients.

Objective:To compare the clinical outcomes of polyetheretherketone (PEEK) and titanium mesh as repair materials in the cranioplasty for cranial defect patients, and to screen the independent factors for postoperative complications of cranioplasty.Methods:A total of 95 patients with cranial defects admitted to our hospital from June 2012 to June 2019 were selected for this study. According to the different repair materials used in cranioplasty, these patients were divided into PEEK group ( n=36) and titanium mesh group ( n=59). General data (hospitalization cost, hospital stay, et al), postoperative complications (intracranial hemorrhage, subcutaneous effusion, infection, seizures, and implant exposure), postoperative plastic satisfaction, and improvement of postoperative neurological function (differences of Glasgow outcome scale [GOS] scores and Mini-mental State Examination [MMSE] scores before and 6 months after surgery) were compared between the 2 groups. Univariate and multivariate Logistic regression analyses were used to screen the independent factors for postoperative complications of cranioplasty. Results:The patients in the PEEK group had significantly higher hospitalization cost but significantly shortened length of hospital stay as compared with those in the titanium mesh group ( P<0.05). The overall incidence of complications in PEEK group was significantly lower than that in titanium mesh group ( P<0.05); the incidence of subcutaneous effusion in PEEK group was significantly lower than that in titanium mesh group ( P<0.05). The PEEK group had significantly higher proportion of patients with good satisfaction in postoperative plasticity, and significantly higher proportions of patients having increased GOS and MMSE scores as compared with the titanium mesh group ( P<0.05). Multivariate Logistic regression analysis showed that repair material was an independent factor for postoperative complications of cranioplasty ( OR=4.550, P=0.019, 95%CI: 1.281-16.161). Conclusion:As compared with titanium mesh, PEEK costs more, but its clinical application effect is better, especially in reducing postoperative complications; selection of appropriate repair materials can be used as one of the methods to reduce postoperative complications.

Objective:To evaluate the clinical efficacy of autologous bone marrow nucleated cell transplantation on ovarian dysfunction-related diseases.Methods:From September 2020 to August 2021, we conducted a before-after study in Shanghai Institute of Planned Parenthood Research Hospital and Shanghai New Hongqiao International Medical Center. A total of 28 patients with premature ovarian insufficiency, perimenopausal syndrome or repeated assisted reproduction failure were treated with ultrasound-guided autologous bone marrow nucleated cell transplantation into the ovaries. Clinical efficacy was evaluated by comparing the patients' self-reported symptoms, endometrial thickness, bilateral ovarian volume, the number of bilateral ovarian follicles, the maximum diameter of bilateral ovarian follicles, and the level of plasma estradiol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, prolactin, and testosterone.Results:After autologous bone marrow nucleated cell transplantation, the number of bilateral ovarian follicles and the maximum diameter of bilateral ovarian follicles statistically significantly increased by an average of 1.26±2.12 ( P=0.005) and (5.40±8.92) mm ( P=0.006), respectively. Endometrial thickness and bilateral ovarian volume increased after treatment although without statistically significance (all P>0.05). However, the ovarian volume, the number of follicles, and the maximum diameter of follicles with greater changes between the left and right sides of the treatment, were found to statistically significantly increase after treatment ( P=0.007, P<0.001, P=0.002). Besides, the levels of FSH, LH, progesterone, and prolactin decreased with no statistical significance (all P>0.05). The stratified analysis of disease types found that endometrial thickness, the number of bilateral ovarian follicles, and the maximum diameter of bilateral ovarian follicles after treatment tended to increase, but only the number of bilateral follicles and the maximum diameter of bilateral ovarian follicles in patients with perimenopausal syndrome showed statistical significance ( P=0.008, P=0.047). Conclusion:The present study suggested that autologous bone marrow nuclear cell therapy could improve ovarian function to some extent, and is expected to be a new treatment method for patients with premature ovarian insufficiency, perimenopausal syndrome, and repeated assisted reproduction failure. However, further studies with larger sample sizes are needed to corroborate the findings.

Objective:To evaluate the clinical efficacy of autologous bone marrow nucleated cell transplantation on ovarian dysfunction-related diseases.Methods:From September 2020 to August 2021, we conducted a before-after study in Shanghai Institute of Planned Parenthood Research Hospital and Shanghai New Hongqiao International Medical Center. A total of 28 patients with premature ovarian insufficiency, perimenopausal syndrome or repeated assisted reproduction failure were treated with ultrasound-guided autologous bone marrow nucleated cell transplantation into the ovaries. Clinical efficacy was evaluated by comparing the patients' self-reported symptoms, endometrial thickness, bilateral ovarian volume, the number of bilateral ovarian follicles, the maximum diameter of bilateral ovarian follicles, and the level of plasma estradiol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, prolactin, and testosterone.Results:After autologous bone marrow nucleated cell transplantation, the number of bilateral ovarian follicles and the maximum diameter of bilateral ovarian follicles statistically significantly increased by an average of 1.26±2.12 ( P=0.005) and (5.40±8.92) mm ( P=0.006), respectively. Endometrial thickness and bilateral ovarian volume increased after treatment although without statistically significance (all P>0.05). However, the ovarian volume, the number of follicles, and the maximum diameter of follicles with greater changes between the left and right sides of the treatment, were found to statistically significantly increase after treatment ( P=0.007, P<0.001, P=0.002). Besides, the levels of FSH, LH, progesterone, and prolactin decreased with no statistical significance (all P>0.05). The stratified analysis of disease types found that endometrial thickness, the number of bilateral ovarian follicles, and the maximum diameter of bilateral ovarian follicles after treatment tended to increase, but only the number of bilateral follicles and the maximum diameter of bilateral ovarian follicles in patients with perimenopausal syndrome showed statistical significance ( P=0.008, P=0.047). Conclusion:The present study suggested that autologous bone marrow nuclear cell therapy could improve ovarian function to some extent, and is expected to be a new treatment method for patients with premature ovarian insufficiency, perimenopausal syndrome, and repeated assisted reproduction failure. However, further studies with larger sample sizes are needed to corroborate the findings.

OBJECTIVE:To investigate the omeprazole off-label use and provide reference for rational drug use in clinic. METHODS:In retrospective survey,1838 omeprazole prescriptions were selected from outpatient department during Jun. 2014 to May 2017 by random number table. According to the latest drug instruction,whether the off-label use or not was determined. Off-label use was determined by reviewing domestic and foreign guidelines,literatures and Micromedex database. RESULTS:Among 1838 pre-scriptions,there were 1750 prescriptions of off-label drug use (95.21％),involving 13 items and 3 types of off-label drug use. Main type of off-label drug use was over-indication medication,involving 1747 prescriptions (96.47％). There was no statistical significance in the incidence of off-label drug use between digestive department and non-digestive department or among physicians at different levels(P>0.05). Among 13 items of off-label drug use,7 items were supported by evidence-based medicine evidence of domestic and foreign guideline,expert consensus/suggestion or literature reports,etc.;among which 1 item of off-label drug use were included in Micromedex grading system and other 6 items had no evidence-based medicine evidence. CONCLUSIONS:The phenomenon of omeprazole off-label use is widespread in outpatient department of the hospital,and some off-label drug use are sup-ported by evidences. There are differences in the quality of those evidences. It is suggested to standardize the off-label drug use to avoid legal risks and guarantee the safety of drug use for patients.

Objective To detect the global level of histone 4 lysine 20 (H4K20) methylation and its relation with DNA damage-repair in peripheral blood cell of arsenic-exposed residents in the coal-contaminated arsenism areas in Guizhou,in order to provide a basis to deepen the interpretation of the role of arsenic in inhibiting DNA damage-repair.Methods Jiaole village in coal-burning-borne arsenism areas in Xingren County of Guizhou was selected as the survey point,and 115 cases of arsenic-exposed residents were selected as the arsenic exposed group on the basis of physical examination.Moreover,53 residents from one village of non-epidemic area neighboring the diseased area were selected as controls.Hair and peripheral blood samples of these subjects were collected,and the histone protein was extracted from the lymphocytes separated from blood samples.The hair samples were digested with microwave digestion instrument,and the hair arsenic content was tested via the inductively coupled plasma-mass spectrometry (ICP-MS) method;the level of H4K20 1,2,3 methylation (H4K20me2,me2,me3) in peripheral blood cell was tested by enzyme-linked immunosorbent assay (ELISA);DNA damage of peripheral blood cell was measured by single cell gel electrophoresis (SCGE).Results The testing results of hair arsenic contents showed that the arsenic levels of hair in arsenic exposed group [0.30 (0.19-0.46)μg/g] were significantly higher than those of control group [0.12 (0.08-0.18) μg/g,F=11.968,P ＜ 0.05].Compared with the control (0.44 ± 0.14,0.99 ± 0.41,1.06 ± 0.33),the level of H4K20me1 (0.60 ± 0.29) in arsenic exposed group was higher (F =2.513,P ＜ 0.05),H4K20me2 (0.75 ± 0.26) was lower (F =4.707,P ＜ 0.05),and H4K20me3 (1.20 ± 0.62) was of no significant difference (F =0.582,P ＞ 0.05).The detecting results of DNA damage of the lymphocytes separated from peripheral blood showed a statistically significant increase (F =9.307,9.457,all P ＜ 0.05) in TailDNA％ and Olivetailmoment in arsenic exposed group [Median (M):10.75,11.69]compared with those of control group (M:2.12,1.16).The correlation analysis indicated that the arsenic levels of hair of subjects were positively correlated with H4K20me1 (r =0.214,P ＜ 0.05) and inversely with H4K20me2 (r =-0.224,P ＜ 0.05);H4K20me1 was positively associated with TailDNA％ (r =0.383,P ＜ 0.05) and Olivetailmoment (r =0.380,P ＜ 0.05);H4K20me2 was inversely associated with TailDNA％ (r =-0.290,P ＜ 0.05) and Olivetailmoment (r =-0.298,P ＜ 0.05).Conclusion H4K20 methylated modification makes a response to arsenic exposure of human body,the alteration of H4K20mel/me2 may participate in regulating DNA damage-repair induced by arsenic in vivo.