Objective The aim of this study was to investigate the effects of morusin on the migration and invasion of human glioblastoma U87 cells,and to explore its mechanism of action.Methods The Cell Counting Kit-8(CCK-8)was used to detect the effect of morusin on proliferation of U87 cells.Wound-healing and Transwell assays were used to detect the effects of morusin on mi-gration and invasion of U87 cells.Western blot was employed to detect the effect of morusin on the expressions of MMP-2 and Vimen-tin in U87 cells.Results The results of CCK-8 assay showed that morusin(1,2,4,and 6μg/mL)could significantly inhibit the pro-liferation of U87 cells compared to the 0 μg/mL morusin(P<0.001).The results of wound-healing assay showed that the migration rates of morusin-treated groups(2,4,and 6 μg/mL)were(20.597±1.225)%,(14.734±1.528)%and(7.811±1.496)%,respec-tively,which were significant lower than that in the 0 μg/mL group(40.566±3.284)%(P<0.001).The results of Transwell assay showed that the invasion number of U87 cells treated with morusin at the concentrations of 2,4,and 6 μg/mL was 85.000±6.557,41.000±6.245,and 13.333±3.215,respectively,which were significant lower than that in the 0 μg/mL group(116.667±14.572)(P<0.001).The results of Western blot showed that the expression of Vimentin in U87 cells increased gradually accompanying with the increase of morusin concentrations,while the expression of MMP-2 decreased gradually accompanying with the increase of morusin concentrations(P<0.001).Conclusion Morusin can effectively inhibit the migration and invasion ability of U87 cells,and its effect is positively correlated with the concentration of morusin within a certain range.

Objective:To compare the effect of a 3D-printed ankle and foot orthosis (AFO) with that of a traditional AFO on the recovery of walking function after a stroke.Methods:Thirty-four hemiplegic stroke survivors were randomly divided into an observation group and a control group, each of 17. Both groups were taught good limb placement and given joint mobility, standing and walking training for 4 weeks wearing either a 3D-printed or a conventional AFO. Walking speed, walking endurance, and dynamic balance were evaluated before and after the experiment using the 10-metre walk test (10MWT), the 6-minute walk test (6MWT), and the timed up and go test (TUGT). Integrated electromyography (iEMG) was also performed on each subject′s bilateral rectus femoris, anterior tibialis, and gastrocnemius muscles during walking, and their healthy and affected side iEMG results were compared to assess the activation of the affected lower limb muscles.Results:After treatment, the 10MWT, 6MWT, and TUGT results of both groups had improved significantly, but the observation group′s average results were then significantly better than those in the control group. The iEMG disparities between the healthy and affected sides had also decreased significantly, but on average the disparities in the observation group were significantly smaller than in the control group.Conclusion:Both types of AFO can effectively improve the walking speed, walking endurance, and dynamic balance of hemiplegic stroke survivors and promote muscle activation in the affected lower limb. A 3D-printed AFO is relatively more effective.

Objective:To evaluate the effect of metformin on ultraviolet A (UVA) -induced photoaging of an immortalized human keratinocytes cell line (HaCaT), and to explore its potential mechanisms.Methods:Cell counting kit 8 (CCK8) assay was performed to evaluate the effect of metformin at different concentrations (0 - 100 mmol/L) on the viability of HaCaT cells, and 10 mmol/L metformin was selected for subsequent experiments. Cultured HaCaT cells were divided into a blank control group (conventional culture), a metformin group (treated with culture medium containing 10 mmol/L metformin), a UVA irradiation group (conventional culture for 24 hours followed by 10 J/cm 2 UVA irradiation) and a metformin + UVA group (treated with culture medium containing 10 mmol/L metformin for 24 hours followed by 10 J/cm 2 UVA irradiation) ; UVA irradiation was performed at a dose of 10 J/cm 2 once a day for 3 consecutive days. After 4-day treatment, cells were collected, the β-galactosidase assay was performed to determine the proportion of senescent cells in each group, 2′, 7′-dichlorodihydrofluorescein diacetate assay to detect levels of intracellular reactive oxygen species (ROS), and the comet assay to detect DNA damage levels. Additionally, some HaCaT cells were divided into the blank control group, metformin group, 1.25 μmol/L dorsomorphin (an adenosine monophosphate-activated protein kinase [AMPK] inhibitor) + metformin group, and 2.5 μmol/L dorsomorphin + metformin group, and cells in the latter two groups were treated with 1.25 and 2.5 μmol/L dorsomorphin respectively for 2 hours, followed by the treatment with 10 mmol/L metformin for 24 hours. Western blot analysis was performed to determine the cellular localization and phosphorylation levels of nuclear factor-erythroid 2-related factor 2 (Nrf2). By using the small-interfering RNA (siRNA) -mediated silencing method, siRNA-Nrf2 was transfected into HaCaT cells to knock down Nrf2 expression (siRNA-Nrf2 group) ; 2.5 μmol/L dorsomorphin-treated HaCaT cells or Nrf2-knockdown HaCaT cells were treated with metformin and UVA irradiation (dorsomorphin + metformin + UVA group, siRNA-Nrf2 + metformin + UVA group, respectively), and the proportions of senescent cells were further calculated in each group. Statistical analysis was carried out by using one-way analysis of variance and two-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:Treatment with different concentrations of metformin for 24 hours could affect the viability of HaCaT cells to varying degrees ( F = 5 206.31, P < 0.001) ; there were no significant differences in the relative survival rates of HaCaT cells between the 10 - 20 mmol/L metformin groups and the control group (0 mmol/L metformin group, all P > 0.05), while the relative cell survival rates were significantly lower in the 25 - 100 mmol/L metformin groups than in the control group (all P < 0.05). After UVA irradiation, HaCaT cells shrank significantly and became narrow and elongated, and the intercellular spaces increased; the relative cell survival rate was significantly lower in the UVA irradiation group (76.13% ± 1.03%) than in the blank control group (100.00% ± 1.24%, LSD- t = 14.86, P < 0.001), but significantly higher in the metformin + UVA group (106.69% ± 2.45%) than in the UVA irradiation group (LSD- t = 11.55, P < 0.001). Moreover, the UVA irradiation group showed significantly increased proportions of senescent cells (45.14% ± 4.98%), intracellular ROS levels (144.61% ± 4.91%), and percentages of DNA in the tail (75.33% ± 1.77%) compared with the blank control group (23.84% ± 1.89%, 55.49% ± 1.57%, 1.88% ± 0.29%, respectively, all P < 0.001), while the metformin + UVA group showed significantly decreased proportions of senescent cells (24.26% ± 1.34%), intracellular ROS levels (58.62% ± 2.17%), percentages of DNA in the tail (15.83% ± 1.23%) compared with the UVA irradiation group (all P < 0.001). Western blot analysis showed that the Nrf2 expression in the cytoplasm was lower in the 10 mmol/L metformin group than in the blank control group, while the phosphorylated Nrf2 expression in the nuclei was higher in the 10 mmol/L metformin group than in the blank control group, suggesting that metformin could effectively induce the phosphorylation of Nrf2 and its nuclear translocation; both the pretreatment with 1.25 and 2.5 μmol/L dorsomorphin could significantly reduce the phosphorylation levels of AMPKα and Nrf2 induced by 10 mmol/L metformin. The proportions of senescent cells in the dorsomorphin + metformin + UVA group and the siRNA-Nrf2 + metformin + UVA group were 67.84% ± 2.74% and 65.94% ± 1.33%, respectively, which were significantly higher than those in the metformin + UVA group (37.76% ± 1.64%, t = 14.45, 13.34, respectively, both P < 0.001) . Conclusion:Metformin may inhibit UVA-induced photoaging of HaCaT cells by activating the AMPK/Nrf2 signaling pathway, scavenging ROS and reducing DNA damage.

Objective:To investigate the value of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid- D-Phe1-Tyr3-Thr8-octreotide (DOTATATE) combined with 18F-FDG total-body PET/CT imaging in the diagnosis and heterogeneity assessment of primary foci of neuroendocrine neoplasms (NEN). Methods:Clinical, imaging and pathological data of 39 patients with pathological diagnosis (30 NEN and 9 non-NEN, 18 males and 21 females, age (54.0±11.4) years) who underwent 1/10 activity 18F-FDG and 1/2 activity 68Ga-DOTATATE total-body PET/CT combined imaging in Zhongshan Hospital, Fudan University from August 2020 to March 2023 were retrospectively analyzed. The NEN primary foci were classified as neuroendocrine tumor (NET) G1, G2, G3, and neuroendocrine carcinoma (NEC). Diagnostic efficacy of combined dual-low activity dual-tracer imaging for NEN primary foci and its value for evaluating tumor heterogeneity were analyzed. Results:The sensitivities, specificities, and accuracies of 68Ga-DOTATATE alone and in combination with 18F-FDG total-body PET/CT for the diagnosis of NEN primary foci were 81.2%(26/32), 7/9, 80.5%(33/41) and 90.6%(29/32), 7/9, 87.8%(36/41), respectively. Ten NET G1 and seven NET G2 lesions showed 68Ga-DOTATATE uptake and no 18F-FDG uptake; two NET G2 lesions showed no 68Ga-DOTATATE uptake but 18F-FDG uptake; and two NET G1 and six NET G2 lesions showed both 68Ga-DOTATATE and 18F-FDG uptake. The radiation doses of 68Ga-DOTATATE, 18F-FDG and a single examination of CT were (1.59±0.50), 0.49(0.44, 0.58) and 11.46(10.53, 12.85) mSv, respectively. Conclusion:Combining total-body PET/CT imaging with dual tracers can effectively diagnose NEN primary foci and assess inter-tumor heterogeneity.

Objective:To explore the feasibility of one-tenth dose 18F-FDG total-body PET/CT (TB PET/CT) in patients with malignant tumors. Methods:A retrospective analysis was carried out on 34 preliminarily diagnosed cancer patients (30 males, 4 females, age (64.0±1.6) years) who underwent one-tenth dose (0.37 MBq/kg) 18F-FDG TB PET/CT examination between April 2020 and September 2022 in Zhongshan Hospital, Fudan University. The raw data were reconstructed into 15 min and initial 2 min PET images (G15 and G2, respectively). A matched cohort of 34 preliminarily diagnosed malignant tumor patients (27 males, 7 females, age (63.3±2.1) years) undergoing full dose (3.70 MBq/kg) 18F-FDG conventional digital PET/CT (C PET/CT) examination with a PET scan rate of 2-3 min/bed position, were analyzed in line with the same pathological types. Signal-to-noise ratios (SNR) of G15, G2 and C PET/CT groups were compared, and based on the pathological results, the detection rates of those 3 groups for lesions were also compared. The χ2 test, independent sample t-test, Mann-Whitney U test, and Wilcoxon rank sum test were used for data analysis. Results:The significant differences in gender, age, body mass index (BMI), blood sugar level and postinjection waiting time between TB PET/CT group and C PET/CT group were not found ( χ2=0.98, t values: 0.08, -1.05, z values: 0.68, 0.41, all P>0.05). The SNR, from G15 to C PET and G2 groups, decreased gradually, which were 16.0(11.3, 20.0), 10.5(8.2, 13.5) and 8.4±0.3 respectively ( z values: 5.09, 3.31, -4.24, all P<0.05). All primary lesions and hepatic metastases were detected by G15 and G2 imaging (100%, 37/37) as well as by C PET/CT (100%, 36/36). The detection rates for lymph node metastasis lesions were 10/15 in the G2/G15 groups, which were higher than the detection rate in the C PET/CT group (64.4%(29/45); χ2=62.03, P=0.002). Conclusion:One-tenth dose 18F-FDG TB PET/CT with a 2-minute acquisition is feasibility in the clinical practice.

Drug metabolism and pharmacokinetics (DMPK) is an important branch of pharmaceutical sciences. The nature of ADME (absorption, distribution, metabolism, excretion) and PK (pharmacokinetics) inquiries during drug discovery and development has evolved in recent years from being largely descriptive to seeking a more quantitative and mechanistic understanding of the fate of drug candidates in biological systems. Tremendous progress has been made in the past decade, not only in the characterization of physiochemical properties of drugs that influence their ADME, target organ exposure, and toxicity, but also in the identification of design principles that can minimize drug-drug interaction (DDI) potentials and reduce the attritions. The importance of membrane transporters in drug disposition, efficacy, and safety, as well as the interplay with metabolic processes, has been increasingly recognized. Dramatic increases in investments on new modalities beyond traditional small and large molecule drugs, such as peptides, oligonucleotides, and antibody-drug conjugates, necessitated further innovations in bioanalytical and experimental tools for the characterization of their ADME properties. In this review, we highlight some of the most notable advances in the last decade, and provide future perspectives on potential major breakthroughs and innovations in the translation of DMPK science in various stages of drug discovery and development.

Objective To evaluate the protective effect of pterostilbene against ultraviolet B (UVB)-induced acute damage in HaCaT cells,and to explore related mechanisms.Methods The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo1ium (MTS) assay and flow cytometry were performed to estimate the proliferative activity and the apoptosis and necrosis rate of HaCaT cells treated with different concentrations of pterostilbene respectively,so as to screen the non-toxic concentration of pterostilbene.HaCaT cells were randomly divided into several groups:normal control group receiving no treatment,UVB group irradiated with 57 mJ/cm2 UVB,3 pterostilbene groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours,3 pterostilbene + UVB groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours followed by UVB radiation.Western blot analysis was conducted to detect changes of the transcription factor NF-E2-related factor 2 (Nrf2) expression in cell nuclei and cytoplasm before and after the treatment with pterostilbene and UVB,quantitative PCR to determine the mRNA expression of catalase and superoxide dismutase in the HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to evaluate the activity of catalase and superoxide dismutase.Results MTS assay and flow cytometry showed that 2.44,4.88 and 9.75 μmol/L pterostilbene had non-toxic effect on HaCaT cells.The protein expression of Nrf2 in the nuclei and cytoplasm in the normal control group was 1.03 ± 0.08 and 1.04 ± 0.11 respectively.Compared with the normal control group,the protein expression of Nrf2 in the nuclei and cytoplasm experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups,and the UVB group showed similar protein expression of Nrf2 in the cytoplasm,but significantly increased protein expression of Nrf2 in the nuclei (1.77 ± 0.08,q =17.24,P ＜ 0.01).Compared with the normal control group and UVB group,the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups all showed significantly lower protein expression of Nrf2 in the cytoplasm (0.86 ± 0.10,0.87 ± 0.11 and 0.46 ± 0.11 respectively,all P ＜ 0.05),but significantly higher protein expression of Nrf2 in the nuclei (2.38 ± 0.11,2.57 ± 0.11 and 2.07 ± 0.13,all P ＜ 0.01).As qPCR showed,UVB radiation could significantly inhibit the mRNA expression of CAT (P ＜ 0.05),but had no obvious effect on the mRNA expression of SOD (P ＞ 0.05).The mRNA expression of CAT and SOD experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups compared with the normal control group (P ＞ 0.05).However,2.44,4.88 and 9.75 μmol/L pterostilbene could significantly reduce the inhibitory effect of UVB radiation on the mRNA expression of CAT (P ＜ 0.05) and up-regulate the mRNA expression of SOD in the pterostilbene + UVB groups (P ＜ 0.05).ELISA revealed that UVB radiation could inhibit the activity of CAT and SOD in the HaCaT cells (both P ＜ 0.001),while 2.44,4.88 and 9.75 μmol/L pterostilbene could reduce the inhibitory effect of UVB radiation on the activity of CAT and SOD (all P ＜ 0.05).However,the activity of CAT and SOD were still lower in the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups than in the normal control group (P ＜ 0.05).Conclusion Pterostilbene can prevent UVB-induced acute damage in HaCaT cells by activating the Nrf2 pathway and up-regulating the expression of the downstream antioxidant enzymes CAT and SOD.

Objective To explore the effects of referral and rehabilitation status of patients with total knee arthroplasty (TKA) under the mode of medical alliance.Methods One hundred and ninety cases with total knee arthroplasty in the secondary rehabilitation hospitals after referral from a tertiary hospital from January 2016 to January 2017 were analyzed, including general demographic data, disease and surgical data, rehabilitation data after referral, hospital day and change of medical expenses of the tertiary hospital and rehabilitation hospitals before and after the promotion of the medical alliance mode.Results After receiving the rehabilitation treatment in secondary rehabilitation hospitals, the range of motion, muscle strength and some other indices of the patients' surgical legs were significantly improved (P<0.05). And the hospital day of the surgical hospital was shorten from (15.05±6.59) d to (12.97±5.24) d, and medical expenses were decreased by 2.18% in total, 4.76% per knee and 14.01% in rehabilitation costs.Conclusions Under the mode of medical alliance, transferring patients with TKA from a tertiary hospital to secondary rehabilitation hospitals in their early postoperative phase can facilitate recovery of the patients, and it can also help to reduce hospital day and medical expenses, thus to optimize the allocation of medical resources.

Objective To explore the influence of autophagy on lipopolysaccharide (LPS) induced acute lung injury (ALI) . Methods Forty‐eight Sprague Dawley (SD) rats were randomly divided into four groups ,12 cases in each group :(1)normal saline control group (NS) ,(2)LPS model group (L) ,(3) LPS and autophagy group (L +A) and (4) LPS and autophagy inhibition group (L+I) .Arterial blood samples was obtained for detecting the blood gas ,including PaO2 ,PaCO2 and pH ,and the lung tissue dry/wet ratio was calculated .The HE staining was used to observe the histopathological changes of lung tissue .Moreover the lung le‐sion score was performed ;the expression of microtubule associated protein ,light chain protein 3b(LC3b) ,myeloperoxidase(MPO) , macrophage inflammatory protein 2(MIP‐2) ,interleukin‐1β(IL‐1β) and tumor necrosis factor‐α(TNF‐α) in serum and bronchoalve‐olar lavage fluid(BALF) was assessed by ELISA .Results Compared with the NS group ,arterial blood PaO2 and pH in the group L were decreased and PaCO2 was increased (P<0 .05);compared with the L group ,the arterial blood PaO2 and pH in the L+A group were increased and PaCO2 was declined (P<0 .01) ,the arterial blood PaO2 and pH in the L+ I group were decreased and PaCO2 was elevated ,the differences were statistically significant (P<0 .01) .The LC3b concentration in serum and BALF in the L group and L+I group was declined ,while MPO ,MIP‐2 ,IL‐1βand TNF‐αconcentrations were increased ,while which in the L+ A group were just the opposite .Conclusion Autophagy plays a improvement and protective effect on LPS induced acute lung injure in rat .

Objective To analyze the clinical characteristics of gravidas with HBV in Nanfang Hospital from 2008 to 2014. Methods 22 906 gravidas were retrospectively investigated. Results The HBsAg positive rates were 11.64% and 6.16% when the gravidas were divided into Cantonese and non-Cantonese groups (χ2 =193.370, P < 0.005). The ALT abnormal rates in HBeAg positive and HBeAg negative gravidas were 17.96% and 6.68% (χ2=62.594, P<0.005). Conclusion The HBsAg positive rate of gravidas in Guangdong and the ALT abnormal rate of HBeAg positive gravidas are higher.