ObjectiveThis study aims to enhance of the farnesyl pyrophosphate（FPP） pool in Saccharomyces cerevisiae by heterologously expressing different farnesyl diphosphate synthases（FPSs） from various plants， thereby increasing the production of terpenoid compounds by the engineered yeast. MethodsRNA from mixed samples of roots， stems， and leaves of seven plants including Arabidopsis thaliana， Rosa rugosa， Artemisia annua， Centella asiatica， Humulus lupulus， Medicago sativa， and Panax ginseng was extracted by column chromatography and reverse transcribed into the first strand of complementary DNA（cDNA）， and based on the transcriptome data of the seven species of plants， sequence-specific primers were designed for CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS， the full-length of the genes was cloned， and the genes were analyzed for bioinformatics in order to construct a pESC yeast shuttle vector. These seven plant-derived FPSs were further heterologously expressed in the previous constructed β-elemene-producing yeast， and the yield of β-elemene was indicated for their catalytic acivities. ResultsThe coding sequences of CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS were all of 1 021 bp in length and encoding 301 amino acids， all of which were similarly related to the endogenous FPS-encoding gene（ERG20） in S. cerevisiae. After heterologous expression， RrFPS was identified as the most effective in catalyzing the synthesis of FPP from isopentenyl pyrophosphate（IPP） and dimethylallyl pyrophosphate（DMAPP）. Compared to the control strains， the RrFPS overexpressed yeast strains YB-1-Rr and YB-3-Rr increased the production of β-elemene by 231.25% and 189.3%， respectively. ConclusionBy comparing the functions of FPS-encoding genes from seven different plant sources， it is determined that the protein encoded by the RrFPS from R. rugosa has the best catalytic ability， which can provide key genetic elements for the construction of engineered yeast strain constructs with high terpenoid production.

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

ObjectiveTo investigate the therapeutic effect and mechanism of coptisine on chronic atrophic gastritis （CAG） in rats based on the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodA CAG rat model was induced by multiple factors， including sodium salicylate， N-methyl-N′-nitro-N-nitrosoguanidine （MNNG）， and irregular feeding. The successfully modeled rats were randomly divided into the model group， folic acid group， and high- and low-dose coptisine groups. The high- and low-dose coptisine groups were given coptisine （50， 10 mg·kg^-1， respectively）， and the folic acid group was given folic acid at 2 mg·kg^-1 for 60 days. The pathological changes were detected by hematoxylin-eosin （HE） staining. The ultrastructure of gastric mucosal cells was observed by electron microscopy. Serum pepsinogen Ⅰ （PGⅠ）， pepsinogen Ⅱ （PGⅡ）， and PGⅠ/PGⅡ ratio （PGR） were detected by immunoturbidimetry. Serum gastrin-17 （G-17） level was detected by radioimmunoassay. The content of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in serum of rats was detected by enzyme-linked immunosorbent assay （ELSIA）. Western blot analysis was used to detect the expression levels of TGF-β₁， PI3K， phosphorylated-Akt （p-Akt）， mTOR， and phosphatase and tensin homolog deleted on chromosome 10 （PTEN） in gastric mucosa. The mRNA levels of TGF-β₁， PI3K， Akt， mTOR， PTEN， microtubule-associated protein light chain 3Ⅱ （LC3Ⅱ）， and Beclin-1 were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the normal group， the model group showed atrophy and reduced number of intrinsic glands in the gastric mucosal tissues， as well as inflammatory cell infiltration. The ultrastructure of gastric mucosal cells in the model group displayed nuclear condensation， reduced and swollen mitochondria， and abnormal structure. The serum levels of G-17， PGⅠ， PGR， and the protein and mRNA levels of PTEN in gastric tissues were significantly lower in the model group （P<0.01）， while serum levels of IL-6， IL-1β， TNF-α， and the protein and mRNA levels of TGF-β₁， PI3K， Akt， and mTOR in gastric tissues were significantly higher （P<0.01）. Compared with the model group， various drug intervention groups showed different degrees of improvement in pathological damage and gastric mucosal cell ultrastructure， significantly increased serum levels of G-17， PGⅠ， and PGR （P<0.05，P<0.01）， and significantly decreased levels of IL-6， IL-1β， and TNF-α （P<0.05，P<0.01）. The high-dose coptisine group significantly downregulated the protein and mRNA levels of TGF-β₁， PI3K， Akt， and mTOR （P<0.05，P<0.01）. ConclusionBerberine has a therapeutic effect on CAG in rats， possibly exerting a protective effect on gastric mucosa by inhibiting inflammation and blocking the PI3K/Akt/mTOR signaling pathway.

Objective To investigate the impact of acacetin(Aca)on angiogenesis in diabetes retinopathy(DR)rats by regulating Hippo signaling pathway.Methods Sixty SD rats were grouped into the control group,the DR group,the Aca low dose group(10 mg/kg),the Aca medium dose group(20 mg/kg),the Aca high dose group(30 mg/kg)and Hippo-YAP signaling pathway inhibitor Verteporfin group(Aca high dose 30 mg/kg+Verteporfin 0.8 pmol/kg),with 10 rats in each group.Except the control group,streptozotocin and high-fat feed were used to construct the DR model.Body weight and fasting blood glucose(FBG)levels of rats were measured.Fluorescein angiography(FFA)was applied to observe retinal angiogenesis and fluorescein leakage.Enzyme linked immunosorbent assay was applied to detect serum levels of vascular endothelial growth factor(VEGF)and angiopoietin-2(Ang-2).Pathological changes of retinal tissue were observed by hematoxylin-eosin staining.Western blot assay was applied to detect expression levels of VEGF,hypoxia-inducible factor 1 alpha(HIF-1α),vascular cell adhesion molecule-1(VCAM-1)and Hippo signaling pathway proteins.Results Compared with the control group,retinal cells of rats in the DR group were arranged in a disordered and loose manner,with neovascularization and a large amount of fluorescein leakage,and body weight,the expression of large tumor suppressor homolog 2(LATS2),p-Yes-associated protein(YAP)were reduced.FBG and expression levels of VEGF,Ang-2,HIF-1α,VCAM-1,YAP,transcription activator with PDZ binding motif(TAZ),TEA domain family member 1(TEAD1)were increased(P＜0.05).Compared with the DR group,retina cells of rats in the Aca low,medium and high-dose groups and the Verteporfin group were arranged neatly,with reduced neovascularization and fluorescence leakage,body weight and the expression levels of LATS2 and p-YAP were increased,and FBG,expression levels of VEGF,Ang-2,HIF-1α,VCAM-1,YAP,TAZ and TEAD1 were reduced(P＜0.05).The effect was more obvious in the Aca high dose group.However,there was no significant difference in each indicator between the Verteporfin group and the Aca high dose group.Conclusion Aca can inhibit angiogenesis and improve retinal pathological damage in DR rats,and its mechanism of action may be related to regulating the Hippo signaling pathway.

Objective To investigate the regulatory effect of poria cocos on gastrointestinal motility in mice. Methods A total of 130 Kunming mice were randomly divided into negative control group, low-dose and high-dose groups of raw poria cocos powder, low-dose and high-dose groups of cooked poria cocos powder, low-dose and high-dose groups of poria cocos surrogate culture powder, low-dose, medium-dose and high-dose groups of poria cocos water extract, and low-dose, medium-dose and high-dose groups of poria cocos alcohol extract, with 10 mice in each group. The animals were administered by gavage for 7 days, once a day. After the last administration, the intestinal propulsion function test and gastric solid emptying test were conducted to observe the regulating effect of poria cocos on gastrointestinal motility of mice. Results Compared with the negative control group, the small intestine propulsion rate in the low-dose group of poria cocos surrogate culture powder was significantly increased (P<0.01). Except the high-dose group of raw poria cocos powder, the other poria cocos groups had higher gastric residual rate (P<0.05). Conclusion Poria cocos does not promote intestinal propulsion of mice under normal physiological condition, but it can inhibit gastric empting and exert a moderating effect on gastrointestinal function in normal mice.

Objective:To investigate the clinical effect and demonstrate the techniques of single position laparoscopic nephroureterectomy.Methods:The clinical data of 84 upper urinary tract urothelial carcinoma patients admitted to the Cancer Hospital Chinese Academy of Medical Sciences from September 2018 to July 2022 were retrospectively analyzed, including 39 males and 45 females, with a median age of (64.9±9.3)years and mean BMI of(24.7±3.4)kg/m 2. The tumor was located on the left side in 47 cases and the right side in 37 cases. All 84 patients received single position laparoscopic nephroureterectomy. According to different treatment methods, they were divided into two groups, including 67 cases undergoing nephrectomy first, and then bladder cuff excision was performed along ureter(traditional group), 17 cases undergoing bladder cuff excision before clamping the ureter below the tumor, and then nephrectomy was performed along the ureter to the head side (modified group). There was no statistically significant in the comparison of age [(65.5±9.4)years vs.(62.7±8.9)years], BMI[(24.9±3.5)kg/m 2vs.(23.9±3.3)kg/m 2], left/right side tumor of(38/29 cases vs. 9/8 cases), tumor location (in renal pelvis or calyx or upper/middle/lower ureter being 46/9/12 cases vs. 13/2/2 cases)and tumor stage(T 1-2/T 3-4: 54/13 cases vs. 15/2 cases) between traditional group and modified group ( P>0.05). The operation time, estimate blood loss, postoperative intestinal function recovery time and postoperative drainage time were recorded and compared. Results:All 84 cases were successfully completed without conversion to open surgery. The mean operation time was (160.4±50.1)min, the mean estimated blood loss was(59.4±24.4)ml, the median postoperative intestinal function recovery time was 1(1, 2)d and the mean postoperative drainage time was (4.8±1.9)d(No drainage tube was placed in 4 patients). No Clavien Dindo >grade 3 complications occured. There was no significant difference in the comparison of operation time [(159.2±52.9)min vs. (164.7±38.1)min], estimate blood loss [(60.5±26.2)ml vs. (55.0±17.5)ml], postoperative intestinal function recovery time [1(1-2)d vs. 2(1-2)d] and drainage removal time [(4.8±1.8)d vs. (5.2±2.0)d] between traditional group and modified group ( P>0.05). The postoperative pathology of 84 cases was urothelial carcinoma, and the pathological results of the resection margin were negative. The median follow-up of 84 upper tract urothelial carcinoma patients was 13(3, 28)months. Five patients were lost to follow-up. In traditional group, 5 patients had bladder tumor recurrence, and 5 patients had distant metastasis. In modified group, no bladder tumor recurrence occurred and 1 patient had distant metastasis. Conclusions:Laparoscopic nephroureterectomy in single position is a safe and effective minimally invasive technique for the treatment of upper urinary tract urothelial carcinoma. Treatment of the bladder cuff excision firstly is more in line with the principle of tumor-free and increase surgical space.

ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription in the treatment of chronic atrophic gastritis （CAG） based on network pharmacology and animal experiments，so as to provide scientific basis for clinical application. MethodThe possible targets and pathways of Xianglian Huazhuo prescription in the treatment of CAG were obtained based on the prediction of network pharmacology. The CAG rat model was induced by sodium salicylate，N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） and hunger and satiety disorder. Then the CAG rats were treated with Xianglian Huazhuo prescription and morodan for 60 days. After administration，the rats were sacrificed，and the content of interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β） and vascular endothelial growth factor （VEGF） in serum was determined by enzyme linked immunosorbent assay（ELISA）. In addition， the protein expression of Bad and Bcl-2 in gastric mucosa was detected by immunohistochemistry （IHC）. ResultA total of 241 active components of Xianglian Huazhuo prescription and 53 core targets were obtained. Xianglian Huazhuo prescription affected multiple biological processes，such as cell proliferation and apoptosis，inflammatory reaction，regulation of DNA metabolism，and cell response to redox，as well as phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt），TNF，mitogen-activated protein kinase （MAPK），cancer and cancer-related signaling pathways. The animal model verification showed that Xianglian Huazhuo prescription lowered the levels of IL-6，TNF-α，IL-1β and VEGF in serum of CAG rats，and reduced the protein expression of Bad and Bcl-2 in gastric tissue. ConclusionXianglian Huazhuo prescription could regulate PI3K/Akt signal pathway and improve gastric mucosal injury in CAG by participating in biological processes such as cell proliferation，apoptosis and inflammation.

Objective To explore the role and mechanism of long non-coding RNA (lncRNA)RP11-316M1.12 in breast cancer cells.Methods Bioinformatics analysis was performed to varify RP11-316M1.12 expression pattern in breast cancer tissues and normal tissues.Quantitative real time polymerase chain reaction (qRT-PCR) assay was used to check the expression level of RP11-316M1.12 in breast cancer cells and tissues.Further,the correlationship between RP11-316M1.12 expression and clinical paremeters of breast cancer was analysed according to RP11-316M1.12 level.RP11-316M1.12 was overexpressed in MCF-7 cells,and RP11-316M1.12 was knocked down in MDA-MB-231 cells.Transwell method was used to detect the invasive ability of these cells.Western blot was used to detect the expression of epithelialmesenchymal transition (EMT) markers in these cells.Results Gene Expression Profiling Interactive Analysis (GEPIA) database suggested that RP11-316M1.12 was highly expressed in breast cancer tissues than that in normal tissues.The similar results were got in 65 cases of breast cancer tissues and 23 cases of normal tissues by qRT-PCR assay.Meanwhile,we found that RP11-316M1.12 was enhanced in breast cancer cells than that in normal epithelial cell and RP11-316M1.12 expression is related to TNM stage and distant metastasis in breast cancer.Transwell assay demonstrated that RP11-316M1.12 significantly enhanced breast cancer cells invasion.Mechanismly,over expression of RP11-316M1.12 can remakably downregulated E-cadherin,enhanced ZEB1 and Vimentin expression in these cells.Conclusions RP11-316M1.12 was enhanced in breast cancer,and RP1 1-316M1.12 could accelerate invasion of breast cancer cells.

Objective To evaluate the relationship between 1H-MRS features of brain glioma and its pathological grade and invasiveness, and the correlation between those features and the expression of VEGF, MMP-9 and uPA.Methods The brain gliomas in 55 patients confirmed by pathology were divided into two groups: high grade and low one.An analysis of intragroup and intergroup differences in some metabolite ratios of 1H-MRS in the tumor parenchyma was conducted.The expression of MMP-9,VEGF and uPA in brain glioma was detected,and the correlation between the above three parameters and changes in metabolite ratios of 1H-MRS were explored.Results There was significant difference and correlation in Cho/Cr and Cho/NAA ratio in the tumor parenchyma between high and low grade(P<0.01).The expression of VEGF and MMP-9 in tumor was significantly different(P<0.01) but with significant correlation(P<0.01),and there was no significant difference and correlation in NAA/Cr ratio and uPA expression(P>0.05).Conclusion VEGF and MMP-9 play an important role in the development of brain glioma.Cho/Cr and Cho/NAA ratios provide an important reference for preoperative grading of brain gliomas and indirectly reflect the expression of VEGF and MMP-9.

Objective To use metabonomics method to study the change of the basic materials of month rhythm of wei qi deficiency syndrome; To find the potential markers so as to provides a new way for the essence of the wei qi deficiency syndrome research.Methods Based on the autumnal equinox in lunar calendar month, the beginning of a month (the first day of lunar August), the middle of a month (the 15th day of lunar August), and the end of a month (the 30th day of lunar August) were set as the three days to draw experimental materials. Two weeks before drawing materials, 20 rats were randomly divided into control group and model group, 10 rats in each group. The model rats were modeled by the stimulus of fatigue combined with coldness and hotness. Control group rats received conventional breeding. The rats in the both groups during the three experiments received decollation and the blood was taken at the 12 o’clock at noon. HPLC-MS was used to detect plasma metabolites, and partial least squares were used to make statistical analysis on the data for comparing plasma metabonomics original data of control group and model group. Possible metabolic markers of wei qi deficiency syndrome were explored, and the potential makers of month rhythm change of wei qi deficiency syndrome were deduced.Results Oleamide, phosphatidyl glycerol, cortisol, proline, dimethyl fumarate, and eicosapentaenoic acid may be potential markers of wei qi deficiency syndrome in the beginning of a month. Sphingosine-1-phosphate, malic acid, cortisol, oleamide, carnitine, eicosapentaenoic acid and dimethyl fumarate may be potential markers of wei qi deficiency syndrome in the middle of a month. Cholesteryl acetate, threonine, cortisol, dimethyl fumarate, oleamide, eicosapentaenoic acid and pyroglutamate may be potential markers of wei qi deficiency syndrome in the end of a month.Conclusion Month rhythm change of wei qi deficiency syndrome may be influenced by oleamide, cortisol, eicosapentaenoic acid, dimethyl fumarate, and aconitic acid, and may be closely related to energy metabolism, meanwhile accompanied by regulation of cell, hormone and nerves.