OBJECTIVE: To investigate the antioxidant and osteogenic induction capabilities of calcium phosphate nanoflowers (hereinafter referred to as nanoflowers) in vitro at different concentrations. METHODS: Nanoflowers were prepared using gelatin, tripolyphosphate, and calcium chloride. Their morphology, microstructure, elemental composition and distribution, diameter, and molecular constitution were characterized using scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive spectroscopy. Femurs and tibias were harvested from twelve 4-week-old Sprague Dawley rats, and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured using the whole bone marrow adherent method, followed by passaging. The third passage cells were identified as stem cells by flow cytometry and then co-cultured with nanoflowers at concentrations of 0, 0.4, 0.8, 1.2, 1.6, 2.0, 2.4, 2.8, 3.2, and 3.6 mg/mL. Cell counting kit 8 (CCK-8) assay was performed to screen for the optimal concentration that demonstrated the best cell viability, which was subsequently used as the experimental concentration for further studies. After co-culturing BMSCs with the screened concentration of nanoflowers, the biocompatibility of the nanoflowers was verified through live/dead cell staining, scratch assay, and cytoskeleton staining. The antioxidant capacity was assessed by using reactive oxygen species (ROS) fluorescence staining. The in vitro osteoinductive ability was evaluated via alkaline phosphatase (ALP) staining, alizarin red staining, and immunofluorescence staining of osteocalcin (OCN) and Runt-related transcription factor 2 (RUNX2). All the above indicators were compared with the control group of normally cultured BMSCs without the addition of nanoflowers. RESULTS: Scanning electron microscopy revealed that the prepared nanoflowers exhibited a flower-like structure; transmission electron microscopy scans discovered that the nanoflowers possessed a multi-layered structure, and high-magnification images displayed continuous atomic arrangements, with the nanoflower diameter measuring (2.00±0.25) μm; energy-dispersive spectroscopy indicated that the nanoflowers contained elements such as C, N, O, P, and Ca, which were uniformly distributed across the flower region; Fourier transform infrared spectroscopy analyzed the absorption peaks of each component, demonstrating the successful preparation of the nanoflowers. Through CCK-8 screening, the concentrations of 0.8, 1.2, and 1.6 mg/mL were selected for subsequent experiments. The live/dead cell staining showed that nanoflowers at different concentrations exhibited good cell compatibility, with the 1.2 mg/mL concentration being the best (P<0.05). The scratch assay results indicated that the cell migration ability in the 1.2 mg/mL group was superior to the other groups (P<0.05). The cytoskeleton staining revealed that the cell morphology was well-extended in all concentration groups, with no significant difference compared to the control group. The ROS fluorescence staining demonstrated that the ROS fluorescence in all concentration groups decreased compared to the control group after lipopolysaccharide induction (P<0.05), with the 1.2 mg/mL group showing the weakest fluorescence. The ALP staining showed blue-purple nodular deposits around the cells in all groups, with the 1.2 mg/mL group being significantly more prominent. The alizarin red staining displayed orange-red mineralized nodules around the cells in all groups, with the 1.2 mg/mL group having more and denser nodules. The immunofluorescence staining revealed that the expressions of RUNX2 and OCN proteins in all concentration groups increased compared to the control group, with the 1.2 mg/mL group showing the strongest protein expression (P<0.05). CONCLUSION The study successfully prepares nanoflowers, among which the 1.2 mg/mL nanoflowers exhibits excellent cell compatibility, antioxidant properties, and osteogenic induction capability, demonstrating their potential as an artificial bone substitute material.
Animals ; Osteogenesis/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Calcium Phosphates/pharmacology* ; Rats, Sprague-Dawley ; Rats ; Antioxidants/chemistry* ; Cells, Cultured ; Cell Differentiation/drug effects* ; Nanostructures/chemistry* ; Tissue Engineering/methods* ; Bone Marrow Cells/cytology* ; Coculture Techniques ; Tissue Scaffolds/chemistry* ; Male ; Biocompatible Materials/chemistry* ; Cell Survival ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Cell Proliferation

Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics* ; Manganese/metabolism* ; Plants, Genetically Modified/genetics* ; Plant Proteins/physiology* ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Cation Transport Proteins/metabolism* ; Stress, Physiological

Since the concept of patient versions of guidelines （PVGs） was introduced into China， several PVGs have been published in China， but we found that there is a big difference between the concept of PVG at home and abroad， and the reason for this difference has not been reasonably explained， which has led to ambiguity and even misapplication of the PVG concept by guideline developers. By analyzing the background and purpose of PVGs， and the understanding of the PVG concept by domestic scholars， we proposed the term patient guidelines （PGs）. This refers to guidelines developed under the principles of evidence-based medicine， centered on health issues that concern patients， and based on the best available evidence， intended for patient use. Except for the general attribute of providing information or education， which is typical of common health education materials， PGs also provide recommendations and assist in decision-making， so PGs include both the patient versions of guidelines （PVG） as defined by the Guidelines International Network （GIN） and "patient-directed guidelines"， i.e. clinical practice guidelines resulting from the adaptation or reformulation of recommendations through clinical practice guidelines.

At present， the process and methodology of patient guidelines （PGs） development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers， we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus， so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage， the construction stage， and the completion stage in general， but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network， i.e.， team formation， screening of recommendations， guideline drafing， user testing and feedback， approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process （forming a team， determining the scope of the PG， searching， evaluating and integrating evidence， forming recommendations， writing the guideline， and expert review）. Based on the different models， we sort out the process and methods of PG development and introduce the specific methods of PG development， including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines， with a view to providing reference for guideline developers and related researchers.

Objective:To investigate the level of serum lipoprotein a [Lp (a)] in patients with diffuse large B-cell lymphoma (DLBCL) and its clinical significance.Methods:A retrospective cohort study was performed. The clinical data of 87 patients with DLBCL who were treated at Changshu No.2 People's Hospital from January 2017 to June 2022 (the newly treated DLBCL group) were retrospectively analyzed, and 78 healthy physical examination subjects were selected as the control group. The level of Lp(a) in the two groups and the level of Lp(a) in DLBCL patients achieving different therapeutic effects after treatment were compared. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of serum Lp(a) in predicting the therapeutic effect of DLBCL patients, and the area under the curve (AUC) was calculated to determine the optimal critical value. Based on the optimal critical value, patients with DLBCL were divided into low Lp(a) group and high Lp(a) group, and the clinicopathological characteristics of DLBCL patients with different Lp(a) levels were compared. Cox proportional hazards model was used to analyze the factors affecting the prognosis of DLBCL patients. Kaplan-Meier method was used to compare the relapse-free survival (RFS) and overall survival (OS) of DLBCL patients with different Lp(a) levels.Results:The level of Lp (a) in the newly treated DLBCL group was higher than that in the control group[ (0.24±0.09) g/L vs. (0.09±0.06) g/L], and the difference was statistically significant ( t = 3.61, P = 0.019). Among 87 patients, 54 achieved complete remission (CR), 23 achieved partial remission (PR), and 10 achieved progression of the disease (PD). The Lp (a) levels of patients achieving CR, PR, and PD were (0.09±0.09) g/L, (0.12±0.08) g/L, and (0.25±0.15) g/L, respectively. The Lp (a) levels in patients achieving CR and PR were lower than those in the newly treated DLBCL patients [(0.24±0.09) g/L], and the differences were statistically significant (all P < 0.05). There was no statistically significant difference in the Lp (a) levels between patients achieving PD and the newly treated DLBCL patients ( P > 0.05). The ROC curve results showed that the optimal critical value of serum Lp (a) in predicting the efficacy of DLBCL patients was 0.25 g/L, AUC was 0.776 (95% CI: 0.676-0.876, P < 0.05), and its sensitivity and specificity was 66.67%, 82.76%, respectively. According to the optimal critical value of Lp (a) (0.25 g/L), patients were divided into the low Lp (a) group (≤ 0.25 g/L) (57 cases) and the high Lp (a) group (>0.25 g/L) (30 cases). The proportion of patients with lactate dehydrogenase level >227 U/L, Ann Arbor stage Ⅲ-Ⅳ, and extranodal organ involvement >1 in the high Lp (a) group was higher than that in the low Lp (a) group, and the differences were statistically significant (all P < 0.05). Cox multivariate analysis results showed that Ann Arbor stage Ⅲ-Ⅳ, international prognostic index (IPI) score 3-5, and Lp (a)>0.25 g/L were independent risk factors for OS in DLBCL patients (all P < 0.05); Ann Arbor stage Ⅲ-Ⅳ and IPI score 3-5 were independent risk factors for RFS in DLBCL patients (all P < 0.05). The median OS in the low Lp (a) group was not reached; the median OS of the high Lp (a) group was 21 months, and there was a statistically significant difference in OS between the two groups ( P = 0.001). The median RFS time was not reached in the low Lp (a) group and the high Lp (a) group; and there was no statistically significant difference in RFS between the two groups ( P = 0.102) . Conclusions:Lp(a) level of DLBCL patients is increased, and Lp(a) could be a factor influencing the prognosis of DLBCL.

Objective:To explore the influencing factors and implementation paths of medical and preventive integration of chronic disease in county medical communities (CMCs), providing references for further promoting the integration of medical and prevention and improving the collaborative mechanism of medical and prevention.Methods:From October to November 2023, based on the principle of geographically balanced sampling, medical staff from 6 leading hospitals and 18 other member units of 6 CMCs in Zhejiang Province were selected as survey subjects. A self-designed survey questionnaire was conducted, mainly including the development and evaluation of chronic disease medical and preventive integration services in CMCs. The service development was designed according to the collaborative symbiosis management model, including 4 dimensions of collaborative symbiosis scenarios, willingness, ability, and process, as well as 11 secondary elements. Using secondary elements as the conditional variables and the integration effect of chronic disease medical and prevention as the outcome variable, a qualitative comparative analysis method was used to explore the relationship between multiple conditional variables and their combinations with high integration effect of chronic disease medical and prevention.Results:571 valid questionnaires were collected, with an effective response rate of 96.62%. The consistency of a single secondary element(including conditional non sets) on the high effectiveness of medical and preventive integration was less than 0.9, which cannot constitute a necessary condition for explaining the outcome variable. The configuration analysis results showed that the consistency of the condition combination formed by the interaction of multiple secondary elements was 0.835-0.845, indicating that the condition combination of multiple elements constituted a sufficient non necessary condition for the high integration effect of chronic disease medical and prevention. The configuration path for achieving high integration of medical and preventive effects could be divided into four categories, among which the feature of scenario-process dominance was to create a perfect service scenario and service process as the main focus; The characteristic of the willingness-process dominant type was to stimulate the service willingness of medical staff and improve the service process as the leading factor; The characteristic of the scenario-willingness-ability dominant type was to create a comprehensive service scenario, stimulate the service willingness of medical staff, and enhance service capabilities as the main focus; The characteristic of the willingness-ability-process dominant type was to stimulate the service willingness of medical staff, enhance service capabilities, and improve the service process as the dominant factor. In addition, the four types of configuration paths mentioned above all covered the two secondary elements of endogenous dynamics and professional competence, with a total coverage of 0.626 and a total consistency of 0.821.Conclusions:The configuration path formed through the interaction of multiple elements can effectively achieve the integration of chronic disease medical and prevention. The CMCs should choose the appropriate configuration path based on the actual situation. In addition, special attention should be paid to the endogenous motivation and professional capacity building of medical staff.

The standardization of classification methods of Traditional Chinese Medicine(TCM) ancient books can provide a clear and reliable reference for all kinds of TCM ancient books collection units, which can also promote the sharing and utilization of TCM ancient books. We studied and investigated the classification methods of TCM ancient books in past dynasties. The standard on classification of TCM ancient books was formulated by compared with the classification table of Zhongguo Zhongyi Guji Zongmu, and referred to the classification table of Zhonghua Guji Zongmu. This standard specified three-level categories and classification principles of TCM ancient books, and mainly composed of basic categories, three-level category table, classification principles and examples, and instructions for use.

Objective:To evaluate the efficacy and safety of acupuncture combined with antihypertensive drugs in the treatment of essential hypertension through meta-analysis.Methods:RCTs about acupuncture combined with antihypertensive drugs in the treatment of essential hypertension were retrieved from CNKI, VIP, WANFANG, CBM, PubMed, Embase and Web of Science from the establishment of the databases to January 28, 2022. The risk and quality of literature publication bias were evaluated according to Cochrane 5.1.0 System Evaluation Manual, and relevant data were extracted. RevMan 5.4 software was used for meta-analysis.Results:A total of 36 RCTs involving 2 905 patients were included. Meta-analysis results showed that compared with antihypertensive drugs, acupuncture alone demonstrated advantages in reducing systolic blood pressure [ SMD=-0.70 (-1.05, -0.36), P<0.01], diastolic blood pressure [ SMD=-0.69(-1.06, -0.32), P<0.01], antihypertensive efficacy [ RR=1.11 (1.04, 1.19), P<0.01], symptomes efficacy [ RR=1.21 (1.11, 1.31), P<0.01] and comprehensive efficacy [ RR=1.35 (1.16, 1.57), P<0.01], without serious adverse reactions. Conclusion:Acupuncture alone has good clinical efficacy and safety in the treatment of essential hypertension, compared with antihypertensive drugs alone. However, researches with large samples and high quality are still needed to support the conclusion.

Objective:To construct a primary health care institutions performance evaluation index system from the perspective of health value orientation under the background of countywide medical alliances construction.Methods:From May 2021 to February 2022, preliminary screening was made on core performance evaluation indexes via literature review; purposive sampling was used to select the dean/vice dean, persons in charge of medical service, and those in charge of public health service responsible for performance evaluation at the community health service center. Then semi-structured interviews were made on the existing performance evaluation and assessment plans as well as existing problems of primary medical and health institutions. Based on the " input-process-output" performance evaluation model, the thematic framework analysis method was used to analyze the interview data, and combined with literature research results, a preliminary performance evaluation index system for primary medical and health institutions was built under the guidance of health value. From March to May 2022, the Delphi expert consultation method was used to evaluate the importance and operability of indexes. The threshold method was used to screen indexes, and analytic hierarchy process was used to calculate the weights of evaluation indexes.Results:The health value oriented performance evaluation index system for primary healthcare institutions included 3 first-level indexes, 9 second-level indexes, and 50 third-level indexes. The first-level indexes were output (0.377 3), input (0.336 3), and process (0.286 4) in descending order of weight. The top three weighted second-level indexes were health manpower(0.177 8), health literacy and health outcomes (0.157 6), as well as responsiveness and satisfaction (0.142 6). The third-level indexes included 17 medical indexes, 16 prevention indexes, and 17 medical prevention integration indexes. The top three weighted indexes for inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and responsiveness of residents seeking medical treatment (0.040 1); The top three weighted third-level indexes excluding inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and surplus funds used for personnel incentives (0.045 5).Conclusions:The performance evaluation index system of primary health care institutions built under the health value orientation is scientific, conducive to promoting the health-orientated transformation and improving the efficiency of primary health care services.