Objective To compare the effects of three kinds of fixative solutions on paraffin section of mouse lens tissue and optimize the fixing?method of paraffin section in mouse lens tissue. Methods Three kinds of conventional fixatives were selected for the test ,including the conven?tional Davison’s solution,modified Davison’s solution and 10%neutral buffered formalin. Mice eyeball tissues were fixed with three different fixa?tives,embedded,sliced and then stained with HE method. The paraffin slices were observed under the light microscope. Results The structures of lens and retina fixed in conventional Davison ’s fixative solutions were clear and intact ,and the cells were arranged regularly and compactly. There was no eyeball distortion,contraction and retinal detachment in the eyeballs fixed in modified and conventional Davison’s fixative solution. However,the ones fixed in 10%neutral buffered formalin showed eyeball distortion and contraction,space and spherules. Conclusion The mice lens slides made from tissues fixed by conventional Davison ’s fixative solution are better than fixed by modified Davison ’s fixative solution and the 10%neutral buffered formalin fixed ones.

Objective To explore the effects of using proton pump inhibitors (PPIs) on the outcomes of hip fracture.Methods Searches were conducted through Medline,Embase,Cochrane Library and Chinese Biomedical Literature Database to identify the studies of the association between PPIs exposure and hip fracture.Quality of studies was assessed using the Newcastle-Ottawa Quality Assessment Scale.Pooled odds ratios (ORs) and 95％ confidence interval (CIs) were calculated for the risk of hip fracture associated with current exposure of PPIs.And several subgroups were analyzed by dosing duration,dose,osteoporosis and corticosteroid usage to explore potential study heterogeneities.All statistical analyses were performed with STATA software.Results Among 11 publications included for final analysis,there were a total of 1 107 577 subjects with an average age of over 60 years.A positive relationship existed between PPIs exposure and hip fracture with an OR of 1.46 (95％ CI:1.26-1.70,P =0.000) as compared with nonPPI-users,especially those on concurrent corticosteroid and PPIs.A significantly increased risk of hip fracture was found in the group of a short-term duration for under 1 year (OR =1.18,95％ CI:1.01-1.38,P =0.041),medium-term for 1-3 years (OR =1.23,95％ CI:1.01-1.49,P =0.038) and longer duration for over 6 years (OR =1.38,95％ CI:1.27-1.50,P =0.000).Furthermore,concurrent use of PPIs was not associated with an increased risk of hip fracture in a definite dose-response manner.As compared with non-PPI-users,no significantly increased risk of hip fracture was found in PPI-users with osteoporosis (P ＞ 0.05).Publication bias was not present.Conclusions Use of PPIs may be somewhat associated with an increased risk of hip fracture.Considering potential adverse effects,clinicians should prescribe cautiously PPIs for high-risk patients,especially elders.

The kidney stores essence,essence generates marrow is one of the important contents in zaag-xiang theory of traditional Chinese medicine (TCM).The kidney-marrow-brain,kidney-marrow-bone and other biological axis put forward by modern scholars took kidney-marrow system as their cores.Derived from this basis as well as under the guidance of TCM holism concept,the construction of TCM kidney-marrow system has been continuously improved;and the kidney-marrow correlation theory has been enriched.To better grasp and apply the kidney-marrow correlation theory can solve clinical problems of major diseases.However,doctors in past dynasties did not clearly define kidney-marrow system. It has not formed a complete theoretical system.Marrow in the kidney-marrow system contained brain,spinal cord and bone marrow.Marrow can generate blood,which nourishes bone together with the bone marrow.The bone marrow connects with the brain and spinal cord,which are all related to blood.Hence,the function of marrow generates blood occupied an important position in the kidney-marrow system. In this paper,the structure and function of the kidney-marrow system were preliminarily explained and constructed;and the kidney-marrow system related theories were further enriched.

There was preliminary understanding of the kidney-marrow correlation from the pre-Qin to Han dynasty.Until Wei-Jin and Sui-Tang dynasty,the close relationship between kidney-essence-marrow-bone-blood-brain was gradually realized in terms of the structure and physiological function.There was a new anatomical understanding of marrow in the Song,Jin and Yuan dynasty.The concept of spinal cord was put forward for the first time.There was also the etiology and pathogenesis on kidney marrow related diseases.In the Ming and Qing dynasty,there was in-depth understanding on the kidney-marrow correlation.However,the concept of kidney-marrow system was not clearly proposed.Modern scholars summarized traditional Chinese medicine (TCM) theories on kidney stores essence,essence generates marrow,marrow enriches brain,nourishes bone and transfers into blood. Then,the kidney-marrow systemhad begun to take form.It laid a theoretical foundation for the construction of kidney-marrow system..

Through reviewing and summarizing classical literatures of traditional Chinese medicine (TCM) and modern medical research results,TCM kidney-marrow system was presented and initially explained.The kidney marrow system is inseparable in structure,interdependent in physiology and mutually influential in pathology.The kidney-marrow system can be applied to the treatment of major diseases in TCM,which included metabolic bone diseases,brain and neurological disorders,haematological diseases and etc.,with significant clinical effects.The in-depth study on TCM kidney marrow system will improve the clinical curative effect of major TCM clinical diseases and achieve the precision treatment in traditional medicine.It displays unique advantages of TCM holism concept and achieves new breakthrough in TCM clinical diseases.

Objective To explore expression and clinical value of VEGF-C and p63 in early esophageal carcinoma and intraepithelial neoplasia. Methods 146 cases were randomized into normal esophageal mucosa, low level intraepithelial tumor, high level intraepithelial tumor and early esophageal carcinoma. The expression of VEGF-C and p63 were detected by using the immunohistochemistry dyeing.Results The expression of VEGF-C immunohistochemistry dyeing had statistical differences among different levels(X~2= 47.455, P <0.001). Normal esophageal mucosa v.s. high level intraepithelial tumor (X~2=36.721, P <0.001), Normal esophageal mucosa v.s. early esophageal carcinoma (X~2=26.483, P <0.001), low level intraepithelial tumor v.s. high level intraepithelial tumor(X~2= 10.025, P<0.0083), low level intraepithelial tumor v.s. early esophageal carcinoma(X~2=16.734, P<0.001). There was a significant correlation between pathological classification and the expression amount of VEGF-C (r = 0.462, P <0.001). The expression of p63 had statistical differences among different levels(X~2=28.962, P <0.05). There was a significant difference on normal esophageal mucosa comparing with low level, high level intraepithelial tumor or early esophageal carcinoma (X~2=12.735, P =0.005, X~2=20.421, P<0.001, X~2=20.854, P<0.001). There was a significant correlation between pathological classification and the expression of p63 (r= 0.272, P<0.05). Conclusion There is a significant correlation in the express of either VEGF-C or p63 comparing with either intraepithelial tumor or early esophageal carcinoma. It may be an early warning indicator.

Objective:To explore the micro RNA (miR)-193b-3p expression in hippocampus tissues of neonatal sepsis rats, as well as its role in neuroinflammatory response and possible mechanisms.Methods:Twenty-four 2-d-old SD rats were randomly divided into control group, lipopolysaccharide (LPS) group, negative control group, and miR-193b-3p group ( n=6); except for the negative control group, the rats in the other 3 groups were intraperitoneally injected with LPS to establish sepsis models; in miR-193b-3p group and NC group, 5 μL miR-193b-3p mimics/controls (20 nmol/L)were injected into the lateral ventricle 3 d before LPS injection. In vitro, PC12 cells were divided into control group, LPS group, miR-193b-3p group, and LPS+miR-193b-3p group; miR-193b-3p mimics were transfected into cells of miR-193b-3p group and LPS+miR-193b-3p group; cells in the LPS group and LPS+miR-193b-3p group were exposed to 100 ng/mL LPS. The miR-193b-3p downstream target gene was analyzed by gene microarray and dual luciferase reporter. Neurobehavioral scale used to assess the neurological function at specific time points in each group. The mRNA expression levels of miR-193b-3p and cytokines (interleukin [IL]-1β, IL-6, tumor necrosis factor [TNF]-α) in the brain tissues or PC12 cells were analyzed by real-time fluorescence quantification PCR (RT-qPCR). The protein expression levels of retinoic acid-related orphan receptor α (RORα), neuronal nuclear antigen (NeuN), ionized calcium binding adaptor molecule-1 (IBA-1) and glial fibrillary acidic protein (GFAP) were detected by double-labelling immunofluorescence. The protein expression levels of RORα in PC12 cells were detected by immunofluorescence staining. Results:(1) Gene microarray and dual luciferase reporter analysis confirmed that RORα was the target gene of miR-193b-3p. (2) As compared with those in rats of the negative control group, the neurobehavioral scores in rats of the LPS group were significantly decreased since 6 h of LPS injection and reached to the lowest at 24 h after LPS ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6, and TNF-α mRNA expression levels in the hippocampus of LPS group were significantly increased, while the miR-193b-3p expression was significantly decreased ( P<0.05). (3) As compared with those in negative control group (3.23±0.92), the neurobehavioral scores in rats of the miR-193b-3p group (7.51±0.84) 48 h after LPS injection were significantly higher ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6 and TNF-α mRNA expression levels in the hippocampus of the miR-193b-3p group were significantly deceased, while the miR-193b-3p expression was significantly higher ( P<0.05). (4) As compared with that in negative control group, the number of NeuN(+)RORα(+) cells in the hippocampus of the LPS group was significantly reduced ( P<0.05); as compared with negative control group, miR-193b-3p group had significantly increased number of NeuN(+)RORα(+) cells in the hippocampus ( P<0.05). (5) As compared with the control group, the LPS group had significantly decreased RORα expression, and significantly increased TNF-α, IL-1β and IL-6 mRNA expression in PC12 cells ( P<0.05). As compared with those in the LPS group, the RORα expression was significantly increased, and the TNF-α, IL-1β and IL-6 mRNA expression levels in PC12 cells were significantly decreased in LPS+miR-193b-3p group ( P<0.05). Conclusion:The miR-193b-3p inhibits the neuroinflammatory response in neonatal sepsis rats by regulating its target molecule RORα mRNA expression in hippocampal neurocyte.

Objective:This study aimed to investigate the physical properties, biocompatibility, and 3D printing performance of a novel hybrid bioink composed of gelatin methacrylated (GelMA) and chitin nanocrystal (ChiNC).Methods:The study was conducted from May 2021 to December 2022, four different bioinks were prepared by adding varying amounts of ChiNC to GelMA bioink. The GelMA concentration in all four bioinks was 100 mg/ml, while the ChiNC concentrations were 0 mg/ml (no ChiNC added), 5 mg/ml, 10 mg/ml, and 20 mg/ml, respectively, named as GC0, GC5, GC10, and GC20 bioinks. The cross-sectional morphology of the hydrogels formed after photocuring the four bioinks was observed using scanning electron microscopy, and the porosity was calculated. Weighing the hydrogels before and after swelling, and then calculate the equilibrium swelling rate. HUVECs were seeded on the surfaces of the hydrogels prepared from the four bioinks and cultured in medium. Cell proliferation was assessed using CCK-8 assays at 1d, 3d, and 7d to compare the proliferation rates of cells on the four hydrogels. HUVECs were added to the four bioinks, and grid-like scaffolds were printed and cultured in medium. Live-Dead staining was performed at 1d and 7d to observe cell viability. Compare the printing effect of bioinks by observing its forming continuous threads properties during extrusion. Finally, tissue-engineered bladder patches simulating the mucosal layer, submucosal layer, and muscular layer anatomical structures of the bladder wall were 3D bioprinted using the optimized bioink composition, and the stability and fidelity of the printed structures were observed to further validate the feasibility of printing multi-layered complex structures with the bioink.Results:Scanning electron microscopy revealed that the porosity of the GC0, GC5, GC10, and GC20 hydrogels were (51.43±6.23)%, (51.85±6.47)%, (50.55±4.59)%, and (42.49±2.20)%, respectively. The differences in porosity between the GC0 group and the other three groups were not statistically significant ( P=0.9994, P=0.9948, P=0.1200). The equilibrium swelling ratio of the other three groups [(8.81±0.41), (7.95±0.19), (7.71±0.14)] was significantly lower than that of the GC0 group (9.37 ± 0.49), and the differences were statistically significant ( P=0.0457, P<0.01, P<0.01). CCK-8 assay showed no significant difference in absorbance value between the GC10 group (0.360±0.009) and the GC0 group (0.357±0.007), GC5 group (0.350±0.012), and GC20 group (0.345±0.018) on the first day ( P=0.9332, P=0.5464, P=0.4937). However, on the third day, the absorbance value of the GC10 group (0.755±0.012) was significantly higher than that of the GC0 group (0.634±0.010), GC5 group (0.704±0.009), and GC20 group (0.653±0.015) ( P<0.01, P=0.0033, P=0.0002). On the seventh day, the absorbance value of the GC10 group (1.001±0.031) was significantly higher than that of the GC0 group (0.846±0.026), GC5 group (0.930±0.043), and GC20 group (0.841±0.024)( P=0.0012, P=0.1390, P=0.0010). The addition of human umbilical vein endothelial cells (HUVECs) into the four groups of hydrogels enabled the printing of grid-like scaffolds, and Live-Dead staining was performed on day 1 and day 7. The cell viability of HUVECs in the four groups on day 1 was (90.13±1.63)%, (90.6±2.45)%, (92.58±2.15)%, and (91.40±3.17)%, respectively. There were no statistically significant differences between the GC0 group and the other three groups ( P=0.9869, P=0.3093, P=0.8008). On day 7, the cell viability was (89.97±3.10)%, (92.18±2.21)%, (92.05±2.25)%, and (90.12±1.97)% for the four groups, respectively. There were no statistically significant differences between the GC0 group and the other three groups ( P=0.3965, P=0.4511, P=0.9995). Bioink extrusion test showed that the GC0 hydrogel could be extruded continuously and form threads at temperatures between 24℃ and 25℃, while the GC10 hydrogel could be extruded continuously and form threads at temperatures between 24℃ and 27℃. Printing tissue engineered bladder patches simulating the anatomical structure of the bladder mucosal layer, submucosal layer, and muscular layer using GC10 bioink, and the printed patches were stable, without collapse, and had high fidelity. Conclusions:Adding ChiNC to GelMA promotes cell adhesion, proliferation, and expands the printing window of GelMA bioink. The biocompatibility of the mixed bioink prepared by adding 10 mg/ml ChiNC in GelMA is good, capable of printing tissue-engineered bladder patches that mimic the anatomical structure of natural bladder walls.

BACKGROUND:There is an increasing demand for orthodontic treatment,and periodontally accelerated osteogenic orthodontics(PAOO)technique can make it possible to move orthodontic teeth that are limited by thin alveolar bone. OBJECTIVE:To investigate the biomechanics of orthodontic tooth movement before and after periodontally accelerated osteogenic orthodontics(PAOO)surgery to increase alveolar bone volume using the three-dimensional finite element method. METHODS:A patient undergoing PAOO surgery before orthodontic treatment to increase bone volume on the labial side of the mandibular anterior region was selected.The patient was under invisible orthodontics.Two three-dimensional finite element models were constructed based on the patient's preoperative and 6-month postoperative cone beam CT data.Both models simulated the movement of tooth 33:experiment Ⅰ:distal-central movement of 0.25 mm;experiment Ⅱ:lingual movement of 0.25 mm;and experiment Ⅲ:intrusion movement of 0.10 mm.The stress distribution and initial displacement trend of tooth 33,periodontal ligament and surrounding alveolar bone under the action of the invisible aligner were analyzed before and after the PAOO procedure. RESULTS AND CONCLUSION:Dental stress analysis:In the same orthodontic tooth movement,the maximum Von-Mises stress and overall stress values of tooth 33 were all higher before surgery than after surgery;there were similar distribution areas of maximum equivalent stress and overall distribution trends of Von-Mises stress before and after surgery.Periodontal ligament stress analysis:In the same orthodontic tooth movement,the maximum Von-Mises stress and overall stress values of the periodontal ligament were higher before surgery than after surgery,and there were similar distribution areas of the maximum equivalent stress and overall distribution trends of Von-Mises stress before and after surgery.Alveolar bone stress analysis:In the same orthodontic tooth movement,the maximum Von-Mises stress values of the alveolar bone around tooth 33 were higher before surgery than after surgery,while the equivalent stress distribution showed a gradual decrease from the top of the alveolar ridge to the root.Initial displacement analysis:In the same orthodontic tooth movement,the initial displacements in the main displacement direction for all six observation points of tooth 33 were smaller before surgery than after surgery,and showed a tendency to gradually decrease from the tooth tip to the apex.Therefore,there were differences in the biomechanical characteristics of orthodontic tooth movement before and after the PAOO surgery.With the clear aligner,the postoperative equivalent stress values on the dentition,periodontal ligament,and surrounding alveolar bone were lower than before the surgery,and the initial displacements of the orthodontic teeth after the surgery are larger than before.These findings suggest that PAOO can release the restriction of thin alveolar bone on the movement of orthodontic tooth by increasing alveolar bone thickness,effectively improving the force on the roots,periodontal ligament,and alveolar bone,avoiding the stress concentration on orthodontic tooth in the thin alveolar bone area that can cause complications when moving,and improving the efficiency of tooth movement.

Objective: To construct ¹³¹I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model. Methods: PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with ¹³¹I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation. Results: The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of ¹³¹I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na¹³¹I), the T/NT ratios significantly increased in ¹³¹I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in ¹³¹I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in ¹³¹I-PAMAM(G5.0)-SR group was higher than that in ¹³¹I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05). Conclusions PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. ¹³¹I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.