ObjectiveTo compare the differences in color， odor， coumarin content and microbial community composition of Angelicae Dahuricae Radix（ADR） during different drying processes， and to explore the correlation between changes in microbial community composition and changes in quality indexes of ADR. MethodsThe fresh ADR was processed at three drying temperatures（50， 70， 100 ℃） by drying and steaming cutting， semi-fresh cutting and drying， fresh cutting and drying， and sulfur fumigation methods. The color values of samples were extracted by Adobe Photoshop 2022 software and subjected to principal component analysis（PCA）， electronic nose was used to identify the odor information of medicinal powders and subjected to loadings analysis， PCA， and linear discriminant analysis（LDA）， and high performance liquid chromatography（HPLC） was used to determine the contents of five coumarins（bergapten， oxypeucedanin， imperatorin， phellopterin， isoimperatorin）. The samples for microbial detection were taken from fresh dried samples， 50 ℃（dried and steamed cut， sulfur fumigated） samples， and 100 ℃（dried and steamed cut） samples when the water content was 50% and 14%， respectively. And the changes of microbial community composition during processing were determined by high-throughput sequencing method. The relationship between the changes of microbial community composition and the changes of odor， color and active component content of ADR during drying process was analyzed by Pearson correlation analysis. ResultsThe color quantification results showed that an increase in drying temperature led to the decrease of brightness value（L）， and the increases of red-green value（a） and yellow-blue value（b）， and the change of processing method had no obvious effect on the color of medicinal materials. The results of odor quantification showed that W1S， W2S， W5S， W2W and W1W sensor were sensitive to the odor changes of ADR and could be used to distinguish ADR decoction pieces from different processing methods. The results of HPLC showed that the coumarin content of ADR decreased with the increase of drying temperature and the delay of processing time， the optimal processing method was drying and steaming cutting method， and the optimal temperature was 50 ℃. High-throughput sequencing results showed that the dominant bacteria in ADR during processing were Achromobacter， Agrobacterium， Nocardioides， Mycobacterium and Enterobacter， the dominant fungi were Coprinopsis， Meyerozyma and Apiotrichum. The results of correlation analysis showed that the quality indexes of ADR were positively correlated with Agrobacterium， Mycobacterium in bacteria， Candida in fungi， and negatively correlated with Bacillus in bacteria. ConclusionThere are significant differences in the color， odor， coumarin content and microbial community composition of ADR in different drying processes， and the best drying method is drying and steaming cutting at 50 ℃. The relative abundance changes of 9 bacterial genera and 4 fungal genera are closely related to the quality formation of ADR during the drying process.

To compare the effects of advanced bone flap and no bone flap on the healing of osteotomy gap in open-wedge high tibial osteotomy (OW-HTO) for the treatment of medial compartment osteoarthritis and knee inversion.Methods:A retrospective study was conducted to analyze the 85 patients who had undergone OW-HTO from March 2021 to December 2021 at Trauma Emergency Centre, The Third Hospital of Hebei Medical University for medial compartment osteoarthritis and knee inversion. The patients were divided into 2 groups according to whether bone flap was grafted. In group A of 42 patients who received advanced bone flap intraoperatively, there were 32 females and 10 males with an age of (63.7±6.6) years; in group B of 43 patients who received no advanced bone flap intraoperatively, there were 31 females and 12 males with an age of (63.2±9.4) years. The measures recorded and compared between the 2 groups included: osteotomy gap healing rates at 3, 6, 12, and 18 months postoperatively; Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Visual Analog Scale (VAS) for pain, and Knee injury and Osteoarthritis Outcome Score (KOOS) at 6 and 18 months postoperatively; medial proximal tibial angle (MPTA) and femorotibial angle at immediate postoperation and 18 months postoperation; postoperative complications.Results:The differences in preoperative general information between the 2 groups were not statistically significant, showing comparability ( P>0.05). The osteotomy gap healing rates at 3, 6, 12, and 18 months postoperatively in group A were significantly higher than those in group B ( P<0.05). The WOMAC (26.1±5.9), VAS (4.1±1.4), and KOOS (47.0±9.7) scores at 6 months postoperatively in group A were significantly lower than those in group B (31.3±8.3, 4.8±1.6, and 56.1±11.9) ( P<0.05), but the differences in the above indicators between the 2 groups at 18 months postoperatively were not statistically significant ( P>0.05). There was no statistically significant difference in MPTA or femorotibial angle at immediate postoperation between the 2 groups ( P>0.05). At 18 months postoperatively, the MPTA in group A (88.7°±1.1°) was significantly better than that in group B (87.7°±1.5°) ( P<0.05). The total complication rate in group B [27.9% (12/43)] was signifcantly higher than that in group A [2.4% (1/42)]( P<0.05). Conclusions:In the OW-HTO treatment of the patients with medial compartment osteoarthritis and knee inversion, application of an advanced bone flap to fill the osteotomy gap can accelerate the gap healing and reduces occurrence of delayed healing or non-healing of the osteotomy gap.

Objective:To analyze the rules of Traditional Chinese Medicine (TCM) for treating diabetes based on the National Patent Database.Methods:In the patent search and analysis platform of the China National Intellectual Property Administration, search the invention patents of the TCM compounds for treating diabetes during January 1,2016-December 31,2020. The Ancient and Modern Medical Records Cloud Platform (V2.3.5) was used to conduct the statistics of Chinese materia medica frequency, the nature and meridian entry, and the analysis of association rules. The cluster analysis and factor analysis were carried out with the SPSS 25.0.Results:A total of 490 TCM compound patents were included, which involve 791 kinds of Chinese materia medica. For each patent, the number of the Chinese materia medica types was rough 13-14 in average. Remarkably, thirty kinds of Chinese materia medica with high frequencies were obtained, where tonic Chinese materia medica accounted for the largest number, followed by the heat-clearing Chinese materia medica. Among these Chinese materia medica the categories of nature are mainly flat, cold and slightly cold, while the flavors are mostly sweet, bitter and pungent, which mainly belongs to the three meridians of lung, kidney and spleen. As for the results of statistical analysis, the association rule analysis indicated that there were 17 effective kinds of Chinese materia medica pairs, such as Rhizoma Dioscoreae-Astragalus Membranaceus and Radix Rehmanniae Recen-Astragalus Membranaceus. Seven Chinese materia medica groups were derived by the cluster analysis, and 11 common factors were extracted through factor analysis. Conclusions:The TCM compounds for treating diabetes are based on clearing heat and moisturizing dryness, nourishing yin and fluid, supplemented by invigorating spleen and removing dampness, dispelling phlegm and dissolving blood stasis, purging fire and detoxification. Notably, in clinical application, phlegm, blood stasis, heat toxin and other pathological products are supposed to be the focus, must identifying the both symptoms and root causes.

Objective:To investigate the clinical characteristics of circadian rhythm disorder of blood pressure and its impact on orthostatic hypotension (OH) in Parkinson′s disease (PD).Methods:A total of 165 PD patients from Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from August 2019 to October 2021 were consecutively enrolled. Medical history and scores of motor and non-motor symptoms of patients were collected. Twenty-four-hour ambulatory blood pressure and OH data were collected, and the OH questionnaire was completed. The incidence of each type of circadian rhythm disorder of blood pressure was investigated. The t test, chi-square test and Mann-Whitney U test were used to determine between-group differences of circadian rhythm disorder of blood pressure. The linear trends in clinical characteristics were tested by linear regression analysis. Logistic regression analysis was used to analyze the relationship between different circadian rhythm disorders of blood pressure and OH as well as symptomatic OH (SOH). Results:In 165 PD patients, the incidence of reverse dipping pattern was 39.39% (65/165), nocturnal hypertension was 43.64% (72/165), and awakening hypotension was 31.52% (52/165). Compared with patients without reverse dipping pattern, patients with reverse dipping pattern were older [(71.72±7.81) years vs (65.29±9.68) years, t=-4.491, P<0.001], had later onset age [(66.67±9.10) years vs (62.16±10.66) years, t=-2.809, P=0.006], longer duration [36.00(20.50, 95.50) months vs 24.00(12.00, 41.75) months, Z=-3.393, P<0.001], higher dose of levodopa (LD) [(426.15±267.38) mg/d vs (284.00±235.58) mg/d, t=-3.590, P<0.001], higher levodopa equivalent dose (LED) [(514.80±360.03) mg/d vs (341.44±284.57) mg/d, t=-3.440, P=0.001], higher Unified Parkinson′s Disease Rating Scale (UPDRS)-Ⅱ scores (12.92±6.38 vs 9.54±5.59, t=-3.434, P=0.001), higher UPDRS-Ⅲ scores (28.34±11.60 vs 21.41±12.18, t=-3.508, P=0.001) and higher percentages of hallucinations [18.46% (12/65) vs 7.00% (7/100), χ2 =5.079, P=0.024]. Compared with patients without awakening hypotension, patients with awakening hypotension were older [(70.83±7.09) years vs (66.44±10.16) years, t=-2.811, P=0.006]. Compared with patients without nocturnal hypertension, patients with nocturnal hypertension had longer duration [39.50(15.00, 96.00) months vs 24.00 (12.00, 36.00) months, Z=-2.944, P=0.003], higher LD [(398.61±251.19) mg/d vs (294.62±254.25) mg/d, t=-2.619, P=0.010], higher LED [(493.28±344.02) mg/d vs (345.05±298.59) mg/d, t=-2.959, P=0.004], higher percentages of hallucinations [19.44% (14/72) vs 5.38% (5/93), χ2 =7.882, P=0.005], higher UPDRS-Ⅱ scores (12.08±6.33 vs 10.00±5.86, t=-2.086, P=0.039), higher UPDRS-Ⅲ scores (26.50±11.72 vs 22.42±12.66, t=-2.034, P=0.044), and greater blood pressure variability (BPV) (20.66±5.47 vs 17.44±5.36, t=-3.798, P<0.001). Trend analysis showed that the variety of circadian rhythm was positively correlated with age and duration, use of levodopa and monoamine oxidase B inhibitors and amantidine, morning and daily LD and LED, UPDRS-Ⅱ, UPDRS-Ⅲ and Hamilton Anxiety Scale scores, hallucinations, OH and SOH, and BPV in PD ( P<0.05). Multivariate Logistic regression analysis showed that awakening hypotension ( OR=3.35, 95% CI 1.55-7.22, P=0.002) and nocturnal hypertension ( OR=2.44, 95% CI 1.20-4.97, P=0.014) were risk factors for OH, and LED ( OR=1.21, 95% CI 1.01-1.43, P=0.035), UPDRS-Ⅲ scores ( OR=1.09, 95% CI 1.02-1.16, P=0.009) and w-BPV ( OR=1.14, 95% CI 1.01-1.29, P=0.029) were independent risk factors for SOH. Conclusions:Circadian rhythm disorder of blood pressure was correlated with age, duration, severity of motor symptoms. Awakening hypotension and nocturnal hypertension are independent risk factors for OH in PD.

OBJECTIVES: This study aimed to clarify the effects of Foxp3 silencing on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLFs) in an inflammatory environment and on cell proliferation and invasiveness, as well as to explore the role of Foxp3 gene in the development of periodontitis. METHODS: An small interfering RNA (siRNA) construct specific for Foxp3 was transfected into hPDLFs. Foxp3 silencing efficiency was verified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the siRNA with the optimum silencing effect of Foxp3 gene was screened. Using lipopolysaccharide to simulate an inflammatory environment in vitro, CCK-8 detected the effect of silencing Foxp3 on hPDLFs proliferation under inflammatory conditions. Wound-healing experiments and transwell assays were conducted to detect the effect of silencing Foxp3 on hPDLF migration under inflammatory conditions. The expression of the inflammatory cytokines interleukin (IL)-6 and IL-8 was detected by RT-PCR and Western blotting under inflammatory conditions. RESULTS: After siRNA transfection, RT-PCR and Western blotting analyses showed that the expression of Foxp3 mRNA in the Foxp3-si3 group decreased significantly (t=21.03, P<0.000 1), and the protein expression of Foxp3 also decreased significantly (t=12.8, P<0.001). In the inflammatory environment, Foxp3 gene silencing had no significant effect on hPDLFs proliferation (P>0.05), and Foxp3 gene silencing promoted hPDLFs migration (P<0.05). Moreover, the expression of IL-6 and IL-8 increased (P<0.05). CONCLUSIONS In an inflammatory environment, Foxp3 gene silencing promoted hPDLFs migration but had no significant effect on hPDLFs proliferation. The expression of inflammatory factors expressed in hPDLFs increased after Foxp3 gene silencing, indicating that Foxp3 gene inhibited inflammation in periodontitis.
Humans ; Cell Proliferation/genetics* ; Cells, Cultured ; Cytokines/metabolism* ; Fibroblasts/metabolism* ; Forkhead Transcription Factors/metabolism* ; Gene Silencing ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Periodontal Ligament/metabolism* ; Periodontitis/metabolism* ; RNA, Small Interfering/metabolism* ; Transcription Factors/metabolism*

Objective : To investigate the effect of transforming growth factor β1 (TGF-β1) on human periodontal ligament fibroblasts (hPDLFs) stimulated by lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS). Methods: hPDLFs were obtained and identified by immunohistochemistry. The stimulating concentration of Pg-LPS was determined by qRT-PCR and CCK-8. The hPDLFs were divided into 4 groups: blank control group, 100 μg/mL pure Pg-LPS; low concentration group, 1 ng/mL TGF-β1+100 μg/mL Pg-LPS; medium concentration group, 10 ng/mL TGF-β1+100 μg/mL Pg-LPS; and high concentration group 100 ng/mL TGF-β1+100 μg/mL Pg-LPS. Cell proliferation was measured by CCK-8 assay at 72 hours, cell migration was measured by scratch and Transwell chamber assays at 24 hours, and the cell cycle of the hPDLFs was measured by flow cytometry at 72 hours. The expression of Forkhead/winged helix transcription factor p3 (Foxp3), interleukin-6 (IL-6) and Epstein-Barr virus-induced gene 3 (EBI3) mRNA in hPDLFs at 72 hours was measured by qRT-PCR, and the expression of Foxp3, IL-6 and EBI3 proteins in hPDLFs at 72 hours was detected by western blot. Results : The immunohistochemistry results showed that anti-vimentin was positive and anti-keratin was negative. At a concentration of 100 μg/mL Pg-LPS, the expression of IL-6 mRNA in hPDLFs was increased (P<0.000 1), and the proliferation of hPDLFs was decreased (P<0.000 1). Therefore, 100 μg/mL PG-LPS was selected to simulate the inflammatory state. 10, 100 ng/mL TGF-β1 could improve the proliferation ability of hPDLFs in inflammatory state (P<0.000 1) ; 1, 10 and 100 ng/mL TGF-β1 could promote the migration ability of hPDLFs in inflammatory state (P<0.000 1). 1, 10 and 100 ng/mL TGF-β1 could accelerate the cell cycle of hPDLFs in inflammatory state (P<0.000 1). 1, 10 and 100 ng/mL TGF-β1 could inhibit the expression of IL-6 gene and protein in hPDLFs in inflammatory state (P<0.000 1), 1 and 10 ng/mL TGF-β1 could increase the expression of EBI3 gene and protein in hPDLFs in inflammatory state (P<0.000 1). 1, 10 ng/mL TGF-β1 could increase the expression level of Foxp3 gene in hPDLFs in inflammatory state, and 10 ng/mL TGF-β1 could increase the expression level of Foxp3 protein (P<0.05). Conclusion TGF-β1 can promote the proliferation and migration of hPDLFs under inflammatory conditions, upregulate EBI3 and inhibit inflammation, which may be related to the expression of the transcription factor Foxp3.

Objective: To examine the association between neonatal birth weight and maternal type 2 diabetes (T2DM), so as to provide insights into the formulation of the early T2DM prevention and improvements of maternal and children health. Methods: Single-nucleotide polymorphisms (SNPs) were collected from two genome-wide association studies (GWAS) that examined the association between neonatal birth weight and maternal T2DM. Inverse variance weighted method was employed for forward Mendelian randomization with 26 birth weight-associated SNPs as instrumental variables and maternal T2DM as the study outcome and for reverse Mendelian randomization with 18 maternal T2DM-associated SNPs as instrumental variables and maternal effects of neonatal birth weight as the study outcome. The heterogeneity was examined using Cochran's Q test, and the pleiotropy was evaluated using MR-Egger regression, while the robustness of the results was evaluated using leave-one-out test. Results: Cochran's Q test detected heterogeneity across two studies (P=0.019, 0.038), and random effect models were employed. Mendelian randomization showed that an increase in neonatal birth weight by per standard error (approximately 488 g) resulted a 29.9% reduction in the risk of maternal T2DM (OR=0.701, 95%CI: 0.547-0.899), and maternal T2DM increased the neonatal birth weight by 0.074 standard errors (OR=1.074, 95%CI: 1.043-1.106). No horizontal pleiotropy was seen for instrumental variables (P=0.241, 0.188). With each SNP excluded in turn, the results of Mendelian randomization study were robust. Conclusion There are bidirectional associations between neonatal birth weight and risk of maternal T2DM.

Objective:To investigate the relationship between bedtime procrastination and daytime sleepiness in college students,and to explore the mediating effect of sleep quality and the moderating effect of gender in the a-bove-mentioned relationship.Methods:A total of 2 823 college students(808 males,2 015 females)from two uni-versities were selected.They were assessed with the Bedtime Procrastination Scale(BPS),Epworth Sleeping Scale(ESS,ESS score≥11 indicated daytime sleepiness)and Pittsburgh Sleep Quality Index(PSQI).Process model 4 was used to test the mediating effect of sleep quality and model 14 was used to test the moderating effect of gender.Results:There were 1 214(43.0％)college students suffering from daytime sleepiness.The scores of ESS in college students were positively associated with the BPS scores(β=0.16).The total scores of PSQI partially mediated the effect of BPS scores on college students'ESS scores,and the value of mediating effect was 39.9％.The association between PSQI scores and ESS scores was moderated by gender(β=-0.13).Conclusion:Daytime sleepiness is associated with bedtime procrastination and sleep quality in college students,and the association be-tween sleep quality and daytime sleepiness is moderated by gender.

Objective:To investigate whether the presynaptic dopamine neuronal depletion in different striatal subregions predicts future development of wearing-off (WO) in Parkinson′s disease (PD) patients.Methods:A retrospective longitudinal study included 57 PD patients who were referred to the Department of Neurology of Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2019 to September 2020, and completed 11C-2β-carbomethoxy-3β-(4-fluorophenyl) tropane dopamine transporter (DAT) positron emission tomography scans at the initial evaluation and received dopaminergic drugs for at least 12 months during follow-up. The time of starting dopaminergic drug treatment and the occurrence of WO were recorded. After adjusting for clinical related factors, the predictive value of DAT uptake and related parameters in striatal subregions for WO was evaluated by Cox proportional hazards model. Results:During a median follow-up period of 23 months, 10 patients (18.18%) developed WO. Patients with WO exhibited less DAT uptake in the caudate nucleus and anterior putamen nucleus (0.66±0.52 vs 1.08±0.42, t=2.76, P=0.008 and 0.66±0.20 vs 0.87±0.28, t=2.27, P=0.027 respectively), especially in these subregions contralateral to the less-affected side of the body, compared to those without WO. Cox proportional hazard models revealed that after adjusting for gender, age, course of disease, baseline Unified Parkinson′s Disease Rating Scale Ⅲ score and increment of levodopa equivalent dosage, the lower the DAT uptake of the caudate ipsilateral to the less-affected side of the body ( HR=0.20, 95% CI 0.07-0.63, P=0.006), as well as the lower the DAT uptake of the caudate nucleus and posterior putamen nucleus ( HR=0.28, 95% CI 0.11-0.69, P=0.006 and HR=0.08, 95% CI 0.01-0.64, P=0.018 respectively) and the higher the ratio of putamen/caudate contralateral to the less-affected side of the body ( HR=2.33, 95% CI 1.02-5.33, P=0.045), the higher the risk of WO. Conclusion:The presynaptic dopamine neuronal loss, particularly bilateral caudate nucleus dopaminergic depletion at the early stage, has predictive value of development of WO in PD.

P-glycoprotein (P-gp) highly expressed in cancer cells can lead to multidrug resistance (MDR) and the combination of anti-cancer drugs with P-gp inhibitor has been a promising strategy to reverse MDR in cancer treatment. In this study, we established a label-free and detergent-free system combining surface plasmon resonance (SPR) biosensor with styrene maleic acid (SMA) polymer membrane proteins (MPs) stabilization technology to screen potential P-gp inhibitors. First, P-gp was extracted from MCF-7/ADR cells using SMA polymer to form SMA liposomes (SMALPs). Following that, SMALPs were immobilized on an SPR biosensor chip to establish a P-gp inhibitor screening system, and the affinity between P-gp and small molecule ligand was determined. The methodological investigation proved that the screening system had good specificity and stability. Nine P-gp ligands were screened out from 50 natural products, and their affinity constants with P-gp were also determined. The in vitro cell verification experiments demonstrated that tetrandrine, fangchinoline, praeruptorin B, neobaicalein, and icariin could significantly increase the sensitivity of MCF-7/ADR cells to Adriamycin (Adr). Moreover, tetrandrine, praeruptorin B, and neobaicalein could reverse MDR in MCF-7/ADR cells by inhibiting the function of P-gp. This is the first time that SMALPs-based stabilization strategy was applied to SPR analysis system. SMA polymer can retain P-gp in the environment of natural lipid bilayer and thus maintain the correct conformation and physiological functions of P-gp. The developed system can quickly and accurately screen small molecule ligands of complex MPs and obtain affinity between complex MPs and small molecule ligands without protein purification.