BACKGROUND:Previous studies have found that neohesperidin can delay bone loss in ovariectomized mice and has the potential to treat osteoporosis,but its specific mechanism of action remains to be explored. OBJECTIVE:To explore the key targets and possible mechanisms of neohesperidin in the treatment of osteoporosis based on bioinformatics and cell experiments in vitro. METHODS:The gene expression dataset related to osteoporosis was obtained from GEO database,and the differentially expressed genes were screened and analyzed in R language.The osteoporosis-related targets were screened from GeneCards and DisGeNET databases,and the neohesperidin-related targets were screened from ChEMBL and PubChem databases,and the common targets were obtained by intersection of the three.The String database was used to construct the PPI network of intersection genes,and the key targets were screened.The DAVID database was used for GO and KEGG enrichment analysis.The AutoDock software was used to verify the molecular docking between the neohesperidin and the target protein.The effect of neohesperidin on osteogenic differentiation of C57 mouse bone marrow mesenchymal stem cells was detected.Complete medium was used as blank control group;osteogenic induction medium was used as the control group;and osteogenic induction medium containing different concentrations of neohesperidin(25,50 μmol/L)was used as experimental group.The expression of alkaline phosphatase,the degree of mineralization,the expression of osteogenic-related genes and target genes during osteogenic differentiation of cells were measured at corresponding time points. RESULTS AND CONCLUSION:(1)9 253 differentially expressed genes,2 161 osteoporosis-related targets,and 326 neohesperidin-related targets were screened.There were 53 common targets among the three.All 53 genes were up-regulated in osteoporosis samples.The PPI network screened the target gene PRKACA of research significance.GO function and KEGG pathway enrichment analysis showed that neohesperidin's treatment of osteoporosis through PRKACA target mainly depended on biological processes such as protein phosphorylation and protein autophosphorylation,acting on endocrine resistance,proteoglycan in cancer,and estrogen signaling pathway to play a therapeutic role.Molecular docking results showed that neohesperidin had a certain binding ability to the protein corresponding to the target PRKACA.(2)The results of alkaline phosphatase staining showed that neohesperidin could promote the expression of alkaline phosphatase in the early stage of osteogenic differentiation of mesenchymal stem cells.Alizarin red staining showed that neohesperidin could promote the mineralization of osteogenic differentiation of mesenchymal stem cells.RT-qPCR results showed that neohesperidin could increase the mRNA expression of alkaline phosphatase,PRKACA,and osteocalcin.(3)These results indicate that neohesperidin may promote osteogenic differentiation through PRKACA target on the estrogen signaling pathway to prevent and treat osteoporosis.

Tumor budding is a distinct pathomorphological feature observed in various types of solid tumor. In recent years， tumor budding has been recognized as an important biological feature associated with tumor invasion and metastasis， and it has become a new focus in the research on tumor progression. Although studies have explored the role of tumor budding in different types of tumor， there are studies in the field of hepatocellular carcinoma （HCC）. This article systematically reviews the research advances in tumor budding in HCC， with a focus on the mechanism of tumor budding， the association between tumor budding and tumor progression， and the potential application of tumor budding in prognostic assessment， in order to provide new insights and strategies for the early diagnosis and treatment of HCC.

BACKGROUND:Calcium phosphate(CaP)coatings are widely used to improve the integration of titanium implants into bone but these coatings are associated with risks of infection.It is thus desirable to confer antibacterial properties to CaP coatings. OBJECTIVE:To prepare CaP-MgO composite coatings by impregnating magnesium oxide(MgO)sol into CaP coatings and assess the in vitro antibacterial activities and cytocompatibility. METHODS:An electrolyte was determined by titration and used for CaP coating electrodeposition on titanium(referred to as Ti-CaP).MgO was impregnated into the coating by immersing in an MgO sol with different mass fractions(15%,30%,50%)and subsequently calcined to form MgO-CaP composite coatings,which were recorded as Ti-CaP-15Mg,Ti-CaP-30Mg and Ti-CaP-50Mg,respectively.Microstructure,tensile properties,critical load,and Mg2+ release of coatings in vitro were characterized.Antibacterial activity was assayed using spread plate method by culturing S.aureus on the pure titanium sheet surface and Ti-CaP,Ti-Cap-15mg,Ti-Cap-30mg and Ti-Cap-50mg surfaces for 24 and 48 hours.Mouse osteoblast suspension was inoculated on pure titanium sheets and Ti-CaP,Ti-CaP-15Mg,Ti-CaP-30Mg and Ti-CaP-50Mg coated titanium sheets,respectively.Cell proliferation was detected by CCK-8 assay,and cell survival rate was calculated.The morphology of composite coating soaked in DMEM was also observed. RESULTS AND CONCLUSION:(1)Homogeneous,microporous CaP coatings consisting of octacaclium phosphate crystal flakes were prepared on titanium by electrodeposition.After sol impregnation-calcination,MgO aggregates were filled into the inter-flake voids.The extent of MgO filling and Mg concentration in the coating increased with the number of sol impregnation procedures.When immersed in phosphate buffered saline,all composite coatings actively released Mg2+ within 1 day;subsequently,the Mg2+ release slowed down on day 3.A small amount of Mg2+ release was still detected on day 7.The yield strength,tensile strength and fracture growth rate of Ti-CaP-30Mg coated titanium were not significantly different from those of pure titanium(P>0.05).There was no significant difference in the critical load of Ti-CaP,Ti-CaP-15Mg,Ti-CaP-30Mg and Ti-CaP-50Mg groups(P>0.05).(2)Except that pure titanium sheet and Ti-CaP had no antibacterial properties,the other samples had good antibacterial properties,and the antibacterial rate increased with the increase of MgO content in the coating.(3)After 1 and 3 days of co-culture,the cell survival rate of Ti-CaP-15Mg,Ti-CaP-30Mg and Ti-CaP-50Mg groups was lower than that of pure titanium group and Ti-CaP group(P<0.05).After 5 and 7 days of culture,there was no significant difference in cell survival rate among five groups(P>0.05).The content of MgO in the coating decreased gradually with the time of immersion in the medium.(4)The MgO sol impregnation added antibacterial properties to the CaP coatings while retained their biocompatibility.

Objective:To explore the relationship between the neural development of preterm infants and gut microbiota.Methods:66 premature infants who were hospitalized in the Neonatology Department of Hunan Children′s Hospital from September 2018 to September 2019 were included in the study. Their fecal samples and clinical data from the first admission were collected. According to the neurodevelopment, the patients were divided into normal neurodevelopment group and neurodysplasia group. The bacterial DNA of fecal samples was extracted by 16S rDNA high-throughput sequencing technology and bioinformatics analysis was conducted to compare the composition and diversity of gut microbiota between the two groups.Results:(1) The Shannon index of gut microbiota in normal neurodevelopmental group and neurodysplastic group was 0.89(0.41, 1.51) and 1.01(0.47, 1.31), respectively. There was no significant difference in diversity index between the two groups ( P>0.05). (2) Bifidobacterium, veronica and negativites in the gut microbiota of the normal neurodevelopmental group were significantly higher (all P<0.05), and streptococcus in the gut microbiota of the dysplastic group were significantly higher ( P<0.05). The gut microbiota of the two groups were mainly enterococcus and escherichia shigella. Conclusions:At the genus level, enterococcus and escherichia are the dominant flora of early gut microbiota in preterm infants. Gut microbiota is related to the neural development of preterm infants. The increased abundance of streptococcus, and the decreased abundance of bifidobacterium, veronicus, and negativites may be risk factors for neurodysplasia of preterm infants. The diversity of gut microbiota in early preterm infants may not be significantly related to neural development.

【Objective】 To explore the efficacy of a comprehensive treatment strategy of bladder hydrodistension, transurethral resection of bladder lesions and triple drug instillation in patients with painful bladder syndrome (BPS). 【Methods】 A total of 15 female BPS patients treated during Jan.2020 and Oct.2021 were enrolled. All patients received bladder hydrodistension and transurethral resection. After operation, intravesical instillation of a triple-drug mixture (2% lidocaine hydrochloride 20 mL, heparin sodium 25 000 U, and dexamethasone 10 mg) was administered once a day for 5 days, and then once per week for 4 weeks. The Visual Analogue Scale (VAS) score, quality of sexual life, health-related quality of life, and self-rating anxiety scale score were compared before treatment and 1, 4 and 12 weeks after treatment. The complications were recorded. 【Results】 All patients completed the treatment. Of 14 patients, the VAS score and self-rating anxiety scale score were lower in week 1, 4 and 12 after treatment, while the health-related quality of life score increased. One patient’s symptoms remained unchanged. The VAS score decreased from (5.47±1.81) to (1.87±1.51) (P<0.05), and the self-rating anxiety scale score decreased from (18.13±8.64) to (6.33±8.22) (P<0.05). The score of health-related quality of life increased from (24.47±5.41) to (31.53±6.49) (P<0.05). 【Conclusion】 The comprehensive strategy is effective in the treatment of bladder pain syndrome, which can relieve pain symptoms and improve patients’ quality of life.

Objective:To compare the differences in lung function between sanitation workers and the general population undergoing routine physical examinations, and to analyze the risk factors for restricted airflow and severity of the condition in sanitation workers.Methods:This study is a large cross-sectional study called "Shanxin Respiratory Health Screening for Ten Thousand People". A total of 1 036 sanitation workers (sanitation group) and 6 701 individuals from the general population undergoing routine physical examinations (control group) were selected as the original study subjects from June 2021 to April 2022 (before matching). Both groups underwent pre-bronchodilator lung function tests, and the differences in lung function characteristics between the two groups were compared. The sanitation group also completed a questionnaire survey. Multivariate and ordinal multinomial logistic regression analysis were used to analyze the risk factors for airflow limitation and its severity.Results:A total of 1 027 individuals from the sanitation group and 999 individuals from the control group were included in the study. There were no significant differences in age, gender, height, weight, and body mass index (BMI) between the two groups (all P>0.05). The rate of airflow restriction was significantly higher in the sanitation group compared to the control group (22.88% vs 8.81%, P<0.001). In the sanitation group, there was no statistically significant difference in a self-assessment test for chronic obstructive pulmonary disease (CAT) scores between individuals with airflow restriction (235 cases) and those without airflow restriction (792 cases) [(1.50±2.50) vs (1.15±2.03) points, P=0.084]. There were no statistically significant differences in forced vital capacity (FVC) as a percentage of predicted value (FVC%pred) between the two groups. However, the sanitation group had significantly lower %pred for forced expiratory volume in one second (FEV 1%pred), FVC/FEV 1 ratio (FEV 1/FVC%pred), forced expiratory flow at 50% of FVC (FEF 50%%pred), forced expiratory flow at 75% of FVC (FEF 75%%pred), and maximal mid-expiratory flow (MMEF%pred) compared to the control group (all P<0.05). The rates of abnormal FEF 50%%pred, FEF 75%%pred, and MMEF%pred were significantly higher in the sanitation group compared to the control group (17.62% vs 10.31%, 17.04% vs 10.01%, 27.26% vs 18.41%, all P<0.001). Small airway parameters and the rate of airflow restriction were significantly higher in past and current smokers of the sanitation group compared to never smokers (all P<0.05). Multifactorial analysis showed that high BMI ( OR=0.929, 95% CI: 0.885-0.974) was a protective factor for airflow restriction, while high smoking index was a risk factor ( OR=1.020, 95% CI: 1.011-1.030). Ordered multinomial logistic regression analysis showed that high BMI ( OR=0.925, 95% CI: 0.882-0.971) was a protective factor for the severity of airflow restriction, while high smoking index ( OR=1.020, 95% CI: 1.011-1.029) was a risk factor for the severity of airflow restriction. Conclusions:The incidences of airflow limitation and small airway abnormalities in sanitation workers are higher than that in general physical examination population. High smoking index and low BMI are independent risk factors for airflow limitation and its severity.

Intranasal administration has the advantages of quick onset, high bioavailability, potential delivery into brain, and non-invasive. However, the limitations of the complex nasal microenvironment, such as the mucosal barrier, ciliary movement, low pH mucous layer, and enzyme degradation, appear to be key challenges in nasal drug development. Although a large number of animal trials provide reference data for the development of nasal drugs, the species differences, high cost and long cycle make this approach more difficult to develop, especially in the high-throughput preclinical screening stage of new drugs. Therefore, it is critical to select a nasal model with good in vitro and in vivo correlation for preclinical drug research. Whether the targeting through the nasal into the brain or blood, the epithelial cell barrier directly determines the bioavailability and efficacy during the intranasal administration. Moreover, mucosal irritation is also one of the critical evaluations of the safety aspects during nasal drugs development, where cell model can be quite useful. Therefore, this review summarized the application of the in vitro nasal model based on nasal epithelial cells in the nasal drug development.

Objective:To explore the therapeutic effect on transposition of peroneal longus tendon in reconstruction of long segment defects of chronic Achilles tendon.Methods:From November 2013 to November 2018, 17 patients with long segment defects of chronic Achilles tendon(greater than 5.0 cm of defect) were admitted in the Department of Trauma and Micro-orthopaedics of Zhongnan Hospital of Wuhan University. The patients were 14 males and 3 females, aged 19-55 years old, with an average of 34.7 years old. Eleven patients had defects on left Achilles tendon and 6 on the right. The chronic defects lasted over 1-12 months with an average of 3.8 months. The broken ends of the Achilles tendon located at 0-5.0 cm from the insertion point. The defects were at 5.0-9.0 cm in length, after debridement of the broken ends. After thoroughly debridement of the scar tissue over the broken end of an Achilles tendon, an autologous peroneal tendon transposition was performed for reconstruction of an Achilles tendon. Postoperative follow-ups were conducted to observe wound healing and complications. Arner Lindholm score was employed to evaluate the therapeutic efficacy before surgery and at the final follow-up. Ankle-hindfoot score of American Orthopedic Foot and Ankle Society(AOFAS) and Achilles Tendon Total Rupture Score(ATRS) were used to evaluate functional recovery. At the final follow-up, heel raise of single affected foot and the maximum circumference of both calves were observed, gait analyses for plantar stress assessment as well as review of ankle CT and MRI scans were conducted for assessment of therapeutic efficacy. Using t test in SPSS 22.0 software to compare preoperative and postoperative AOFAS and ATRS scores, P<0.05 indicates a statistically significant difference. Results:Wounds healed in one stage, except 1 diabetic patient who had haematoma in the wound after surgery and healed after debridement combined with VSD treatment. All the 17 patients had completed the postoperative follow-up that lasted for 24-48 months, with an average of 33 months. According to Arner Lindholm score for efficacy evaluation, 11 patients were in excellent and 6 in good. The postoperative AOFAS scores increased from 62.94 ± 6.51 before surgery to 93.71 ± 6.15 after the surgery. ATRS increased from 29.00 ± 3.54 before surgery to 92.29 ± 3.02 after the surgery, and the score difference between before and after surgery was staistically significant ( P<0.05). No patient experienced a complication such as Achilles tendon rupture, wound infection or nerve injury. During follow-up, ankle functions recovered well, valgus was found powerful as well as heel raise on single foot. The circumference of the affected calf was smaller than that of the healthy side by no more than 1.5 cm. Gait analysis showed balanced distribution of plantar stress. No corpus callosum formation was found in both of the affected and the healthy feet. No obvious sign of osteoarthritis of ankle showed review in CT scans. Ankle MRI scans showed good continuity of the reconstructed Achilles tendons, which become significantly thicker after the surgery. Conclusion:The transposition of the long peroneal tendon is a relatively simple and effective surgical technique for reconstruction of a chronic Achilles tendon rupture or defect in a longer length or a rupture at the insertion point.

Objective:To compare the characteristics and differences of intestinal flora in premature infants with late-onset sepsis (LOS) and pneumonia by high-throughput sequencing technology, and to investigate the relationship between intestinal flora and LOS.Methods:Through the case-control method, premature infants with late-onset sepsis who were hospitalized in the neonatal department of Hunan Children′s Hospital from August 2018 to October 2019 were selected as the case group ( n=8). At the same time, premature infants diagnosed with pneumonia were selected as the control group ( n=8). The fecal samples of 16 premature infants were collected for the first time, and the DNA was extracted. The DNA of the target region was amplified by polymerase chain reaction(PCR). High-throughput sequencing was performed using NovaSeq 6000 platform to analyze the composition and diversity of intestinal flora between the two groups. Results:(1) Alpha diversity analysis showed that there was no significant difference in the richness and diversity of intestinal flora between the two groups(all P>0.05). (2) The intestinal flora in premature infants of LOS group and control group were dominated by Firmicutes and Proteobacteria, and facultative anaerobes such as Enterococcus and Escherichia-Shigella were the dominant flora at the genus level. Metastas statistical analysis showed that there was no statistically significant difference in flora composition between the two groups at the phylum level ( P>0.05). (3) Metastas statistical analysis was carried out at the level of class, order, family, genus, and species. The relative abundance of actinomycetes, digestive streptococcaceae and Clostridium in LOS group was higher than that in pneumonia group, and the difference was statistically significant (all P<0.05). (4) The relative abundance of Staphylococcus in the LOS group was significantly greater than that in the control group, but Metastas statistical analysis showed that there was no statistically significant difference in the relative abundance of staphylococcus between the two groups ( P>0.05). (5) Among the 8 cases of LOS, 3 premature infants had positive blood cultures, namely Streptococcus agalactiae, Streptococcus mitis, and Enterococcus faecalis. Enterococcus faecalis belongs to the genus Enterococcus, and Enterococcus belongs to the dominant genus in the LOS group. Conclusions:Different site infections have effects on intestinal microecology of premature infants. There were differences in intestinal flora composition between premature infants with LOS and premature infants with pneumonia.

OBJECTIVE: To assess the practical and health economical values of non-invasive prenatal test (NIPT) in Changsha Municipal Public Welfare Program. METHODS: A retrospective analysis was carried out on 149 165 women undergoing NIPT test from April 9, 2018 to December 31, 2019. For pregnant women with high risks, invasive prenatal diagnosis and follow-up of pregnancy outcome were conducted. The cost-benefit of NIPT for Down syndrome was analyzed. RESULTS: NIPT was carried out for 149 165 pregnant women and succeeded in 148 749 cases (99.72%), for which outcome were available in 148 538 (99.86%). 90% of pregnant women from the region accepted the screening with NIPT. 415 (0.27%) were diagnosed as high risk. Among these, 381 (91.81%) accepted amniocentesis, which led to the diagnosis of 212 cases of trisomy 21 (PPV=85.14%), 41 cases with trisomy 18 (PPV=48.81%) and 10 cases with trisomy 13 (PPV=20.83%). The sensitivity and specificity of NIPT for trisomy 21, trisomy 18 and trisomy 13 were (97.70%, 99.98%), (97.62%, 9.97%) and (100%, 99.97%), respectively. In addition, 213 and 30 cases were diagnosed with sex chromosomal aneuploidies (PPV=46.2%) and other autosomal anomalies (PPV=16.57%), respectively. For Down syndrome screening, the cost and benefit of the project was 120.79 million yuan and 1,056.95 million yuan, respectively. The cost-benefit ratio was 1: 8.75, and safety index was 0.0035. CONCLUSION NIPT is a highly accurate screening test for trisomy 21, which was followed by trisomy 18 and sex chromosomal aneuploidies, while it was less accurate for other autosomal aneuploidies. The application of NIPT screening has a high health economical value.
Aneuploidy ; Cost-Benefit Analysis ; Female ; Humans ; Noninvasive Prenatal Testing ; Pregnancy ; Retrospective Studies ; Trisomy 18 Syndrome/genetics*