·AIM:To determine the effect of anterior-posterior joint surgery on choroidal thickness in patients with proliferative diabetic retinopathy (PDR). · METHODS: A retrospective, case - control study enrolled 60 eyes of 60 patients with PDR diagnosed at Qingdao Municipal Hospital. The patients, who had conditions that warranted anterior - posterior joint surgery,were divided into a clinically significant macular edema group (PDR/CSME+;31 patients,31 eyes) and a non-CSME group (PDR/CSME-;29 patients,29 eyes). Twenty-seven eyes of 27 normal patients were included in the control group. All affected eyes underwent anterior - posterior joint surgery. After surgery, the subfoveal choroidal thickness (SFCT), and the nasal choroidal thickness (NCT) and temporal choroidal thickness (TCT), which were obtained at a distance of 1500μ m from the fovea in the nasal and temporal directions, respectively, were measured in the control and PDR groups by enhanced depth imaging spectral domain optical coherence tomography (EDI-SDOCT) at 1wk,1,3, and 6mo after surgery. Changes in choroidal thickness after anterior - posterior joint surgery were compared between the groups. ·RESULTS:The SFCT,NCT,and TCT were significantly thicker at 1mo than at 1wk, 3, and 6mo after surgery in the PDR/CSME+ and PDR/CSME- groups(P<0.05). The SFCT, NCT, and TCT were significantly thinner at 6mo than at 1wk,1,and 3mo after surgery in the PDR/CSME+and PDR/CSME- groups(P<0.05). The SFCT,NCT,and TCT in the PDR/CSME+ and PDR/CSME- groups at 1wk, 1, and 3mo after surgery were significantly thicker than those in the control group (all P<0.05), but the SFCT, NCT, and TCT at 6mo after surgery showed no significant difference compared with the control group (all P>0.05). There was no significant difference in the SFCT,NCT, or TCT at 1wk, 1, 3, or 6mo between the PDR/CSME+ and PDR/CSME- groups (P>0.05). ·CONCLUSION:The choroidal thickness of PDR patients increases within 1mo after surgery, and decreased after 1mo,but is not significantly different between the control group and the PDR groups at 6mo after surgery. Whether PDR is associated with CSME has no effect on the choroidal thickness after surgery.

OBJECTIVESTo examine the gene expression profile of bone morphogenetic protein 2 (BMP-2) and vascular endothelial growth factor (VEGF) during entochondrostosis of mice and explore the expression rules and effects between BMP-2 and VEGF, and to detect the expression of VEGF in BMP-2 induced entochondrostosis in vivo.

METHODScDNA microarray technique with 34,000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of BMP-2 and VEGF was performed with GCOS1.2 software. An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in BMP-2 induced entochondrostosis in vivo.

RESULTSThe expression signals of VEGF mRNA and VEGF appeared in cytoplasm during condensation of mesenchymal cell. As the mesenchymal cells differentiated into precartilage, the expression signals decreased in mesenchymal cells, but increased in chondrocytes and kept getting denser in the process of cartilage maturity. The peak expression of VEGF mRNA and VEGF in the experimental group appeared on the 14th day, accompanied by numerous hypertrophic chondrocytes. When mature cartilage calcified and new bone trabecula formed, the expression of VEGF mRNA and VEGF decreased in chondrocytes, but still expressed moderately in the osteoblasts and osteocytes.

CONCLUSIONSThe finding reveals a complex pattern of gene coexpression of BMP-2 and VEGF during the critical period of entochondrostosis. It's feasible for the clinical application of BMP-2 in orthopedics.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cell Differentiation ; genetics ; Chondrocytes ; cytology ; metabolism ; Gene Expression ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Male ; Mice ; Oligonucleotide Array Sequence Analysis ; Osteoblasts ; cytology ; metabolism ; Osteogenesis ; genetics ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate the effects and mechanism of radiation-sterilized allogeneic bone sheets in inducing vertebral plate regeneration after laminectomy in sheep.

METHODSTwelve adult male sheep (aged 1.5 years and weighing 27 kg on average) provided by China Institute for Radiation Protection underwent L3-4 and L4-5 laminectomy. Then they were randomly divided into two groups: Group A (n=6) and Group B (n=6). The operated sites of L4-5 in Group A and L3-4 in Group B were covered by "H-shaped" freeze-drying and radiation-sterilized allogeneic bone sheets (the experimental segments), while the operated sites of L3-4 in Group A and L4-5 in Group B were uncovered as the self controls (the control segments). The regeneration process of the vertebral plate and the adhesion degree of the dura were observed at 4, 8, 12, 16, 20 and 24 weeks after operation. X-ray and CT scan were performed in both segments of L3-4 and L4-5 at 4 and 24 weeks after operation.

RESULTSIn the experimental segments, the bone sheets were located in the anatomical site of vertebral plate, and no lumbar spinal stenosis or compression of the dura was observed. The bone sheets were absorbed gradually and fused well with the regenerated vertebral plate. While in the control segments, the regeneration of vertebral plate was not completed yet, the scar was inserted into the spinal canal, compressing the dura and the spinal cord, and the epidural area almost disappeared. Compared with the control segments, the dura adhesion degree in the experimental regenerated segments was much milder (P less than 0.01), the internal volume of the vertebral canal had no obvious change and the shape of the dura sack remained well without obvious compression.

CONCLUSIONSFreeze-drying and radiation-sterilized allogeneic bone sheets are ideal materials for extradural laminoplasty due to their good biocompatibility, biomechanical characteristics and osteogenic ability. They can effectively reduce formation of post-laminectomy scars, prevent recurrence of post-laminectomy spinal stenosis, and induce regeneration of vertebral plates.

Animals ; Bone Transplantation ; methods ; Laminectomy ; methods ; Regeneration ; Sheep ; Spinal Stenosis ; prevention & control ; Spine ; physiology ; Transplantation, Homologous

Objective To observe the therapeutic effect of fourteen bone-setting manipulations and small splint fixation combined with No.8 Orthopedics Prescription(mainly composed of Rehmanniae Radix,Salviae Miltiorrhizae Radix et Rhizoma,Persicae Semen,Caulis Akebiae,Carthami Flos,Corydalis Rhizoma,and Notoginseng Radix et Rhizoma)on distal radius fracture.Methods A retrospective study was carried out in the analysis of the clinical data of 124 patients with distal radius fractures treated by fourteen bone-setting manipulations and small splint fixation in the Department of Orthopedics,Foshan Hospital of Traditional Chinese Medicine from September 2021 to September 2023.The patients were divided into an observation group(63 cases)and a control group(61 cases)depending on the medication of No.8 Orthopedics Prescription or not.The control group was treated with fourteen bone-setting manipulations and small splint fixation,while the observation group was treated with fourteen bone-setting manipulations and small splint fixation combined with No.8 Orthopedics Prescription.The two groups were treated for s one month and then were followed up for more than six months.The changes in the range of motion(ROM)of wrist pronation and supination and palmar flexion in the two groups were observed before treatment and three months after treatment.The time for starting wrist function exercise,time for subsiding swelling of the affected limb and fracture healing time were compared between the two groups.After six months of treatment,the wrist function improvement effect of the two groups was evaluated.Results(1)After treatment,the time for starting wrist joint functional exercise,time for subsiding swelling of the affected limb and the time for fracture healing in the observation group were significantly shortened compared with those in the control group,and the differences were statistically significant between the two groups(P＜0.01).(2)After three months of treatment,the ROM of wrist pronation and supination and palmar flexion in the two groups were significantly improved compared with those before treatment(P＜0.05),and the improvement of ROM of wrist pronation and supination and palmar flexion in the observation group was significantly superior to that in the control group,the differences being statistically significant(P＜0.01).(3)After six months of treatment,the evaluation of the wrist joint function of the two groups showed that the excellent and good rate of the observation group was 55.56％(35/63),and that of the control group was 45.90％(28/61).There was no significant difference in the improvement of wrist function between the two groups(Z=1.075,P=0.282).Conclusion Both methods can achieve satisfactory efficacy in the treatment of distal radius fracture,and the wrist function of the patients has been effectively restored.The treatment of fourteen bone-setting manipulations and small splint fixation combined with No.8 Orthopedics Prescription can significantly shorten the time for subsiding swelling of the affected limb,promote fracture healing and bone regeneration,improve wrist function,and relieve the pain of patients.

Objective:To investigate the effects of ATG5 and ATG7 genes on the sensitivity of multiple myeloma(MM)cell line RPMI-8226 cells to ferroptosis.Methods:CRISPR/Cas9 technology was used to knock out the autophagy key genes ATG5 and ATG7 in RPMI-8226 cells.Western blot was used to identify gene knockout cells,and detect the expression changes of autophagy-related proteins P62 and LC3B.Flow cytometry was used to detect the change of sensitivity of gene knockout cells to RSL3.The content of intracellular ferrous ions and reactive oxygen species(ROS)level in gene knockout cells were detected.Results:Western blot result confirmed that ATG5 and ATG7 genes were knocked out successfully in RPMI-8226 cells.The result of flow cytometry showed that the cell viability of RPMI-8226 cells was dose-dependent on different concentrations of RSL3(r=-0.969).RSL3(10 μmol/L)was used to induce ferroptosis in cells of control group and gene knockout groups,then the cell viability in gene knockout groups were both higher than control group after 48 hours(both P＜0.001).After knocking out the ATG5 and ATG7 genes,the content of intracellular Fe2+decreased significantly compared with control group(both P＜0.01),and the ROS level also decreased(both P＜0.001).Conclusion:Knockout of ATG5 and ATG7 genes can inhibit the ferroptosis of MM cells,and LAP pathway may be involved in the regulation.

OBJECTIVE: To explore the cytidine-phosphate-guanosine (CPG) sites associated with fas-ting plasma glucose (FPG) and glycated haemoglobin (HbA1c) in twins. METHODS: In the study, 169 pairs of monozygotic twins were recruited in Qingdao, Zhejiang, Jiangsu, Sichuan and Heilongjiang in June to December of 2013 and June 2017 to October 2018. The methylation was detected by Illumina Infinium HumanMethylation450 BeadChip and Illumina Infinium MethylationEPIC BeadChip. According to the Linear Mixed Effect model (LME model), fasting plasma glucose and HbA1c were taken as the main effects, the methylation level (β value) was taken as the dependent variable, continuous variables, such as age, body mass index (BMI), blood pressure, components of blood cells, surrogate variables generated by SVA, and categorical variables, such as gender, smoking and drinking status, hypoglycemic drugs taking, were included in the fixed effect model as covariates, and the identity numbers (ID) of the twins was included in the random effect model. The intercept was set as a random. Regression analysis was carried out to find out the CpG sites related to fasting blood glucose or HbA1c, respectively. RESULTS: In this study, 338 monozygotic twins (169 pairs) were included, with 412 459 CpG loci. Among them, 114 pairs were male, and 55 pairs were female, with an average age of (48.2±11.9) years. After adjustment of age, gender, BMI, blood pressure, smoking, drinking, blood cell composition, and other covariates, and multiple comparison test, 7 CpG sites (cg19693031, cg01538969, cg08501915, cg04816311, ch.8.1820050F, cg06721411, cg26608667) were found related to fasting blood glucose, 3 of which (cg08501915, ch.8.1820050f, cg26608667) were the newly found sites in this study; whereas 10 CpG sites (cg19693031, cg04816311, cg01538969, cg01339781, cg01676795, cg24667115, cg09029192, cg20697417, ch.4.1528651F, cg16097041) were found related to HbA1c, and 4 of which(cg01339781, cg24667115, cg20697417, and ch.4.1528651f) were new. We found that cg19693031 in TXNIP gene was the lowest P-value site in the association analysis between DNA methylation and fas-ting plasma glucose and HbA1c (P_FPG=2.42×10^-19, FDR_FPG<0.001; P_HbA1c=1.72×10^-19, FDR_HbA1c<0.001). CONCLUSION In this twin study, we found new CpG sites related to fasting blood glucose and HbA1c, and provided some clues that partly revealed the potential mechanism of blood glucose metabolism in terms of DNA methylation, but it needed further verification in external larger samples.
Adult ; Blood Glucose ; CpG Islands ; DNA Methylation ; Epigenesis, Genetic ; Fasting ; Female ; Glycated Hemoglobin A ; Humans ; Male ; Middle Aged ; Twins, Monozygotic

Moutan Cortex is an important traditional Chinese medicine, "Fengdan Pi" was known as Dao-di herbs from the root bark of Paeonia suffruticosa cv. Feng Dan for its extracted various active components. However, the genetic basis for their activity is virtually unknown. The transcriptome of the root bark from "Fengdan" was sequenced using the Illumina HiSeq 4000 sequencing platform. The clean reads were then de novo assembled into 72 997 unigenes. Among them, the number of unigenes which could been annotated by dataset Nr and GO was 41 139 and 34 592. The 20 016 unigenes could been annotated by KEGG dataset, which were involved in 5 major categories, 34 middle categories, and 352 metabolism pathways. The number of unigenes which were mapped to the phenylpropanoid biosynthesis pathway, terpenoid backbone biosynthesis pathway, terpenoid biosynthesis pathway, alkaloid biosynthesis pathway, and flavonoid biosynthesis pathway was 214, 104, 152, 55 and 36 respectively, suggesting that they are involves in these pathways of pharmaceutically important. Furthermore, there also showed remarkable differences in groups which enrichment ratio of the different expressed gene compared. In addition, a total of 9 939 SSRs were identified from the sequence of 72 997 unigenes. This study not only provides many valuable basal data which was important gene in the synthesis pathway of secondary metabolites with gene searching, but also has important significance to find molecular marker in germplasm for breeding and improvement.

OBJECTIVETo determine whether additional Chinese medicine (CM) could prolong survival and improve the quality of life (QOL) in patients with advanced non-small cell lung cancer (NSCLC) compared with Western medicine (WM) alone.

METHODSThis was a multicenter, prospective cohort study. A total of 474 hospitalized patients with stage III-IV NSCLC were recruited and divided into 2 groups. Patients in the WM group received radiotherapy, chemotherapy, and optimal supportive therapy according to the National Comprehensive Cancer Network (NCCN) guidelines. In the integrative medicine (IM) group, individualized CM (Chinese patent medicines and injections) and WM were administered. The primary end point was overall survival, and the secondary end points were time to disease progression, adverse events, and QOL. Follow-up clinical examinations and chest radiography were performed every 2 months.

RESULTSThe median survival was 16.60 months in the IM group and 13.13 months in the WM group (P<0.01). The incidences of loss of appetite, nausea, and vomiting in the IM group were significantly lower than those in the WM group (P<0.05). The QOL based on Functional Assessment of Cancer Therapy-Lung in the IM group was markedly higher than that in the WM group at the fourth course (P<0.05).

CONCLUSIONSAdditional CM may prolong survival and improve the QOL patients with NSCLC. The adverse effects of radio- and chemotherapy may be attenuated as CM is used in combination with conventional treatments.