Transplantation of microencapsulated genetic engineered stem cells is a new biological approach for the treatment of many refractory diseases.Transplantation of microencapsulated genetic engineered stem cells can transfect the interest gene into the target cells and further implants the gene to host after being microencapsulated.The microcapsules improve the survival time of target cells and remain the cells to play their biological function for a longterm duration,and the microencapsulated cells present with immunoisolation and good biocompatibility after transplantation.This technology will make it possible for the xenotransplantation or gene engineered cell transplantation.Recently,transplantation of microencapsulated genetic engineered stem cells has been used in clinical trial of retinal pigment degeneration and made progress,which laid a foundation for the management of other chronic and refractory eye diseases.The concept and principle of the microencapsulation technology are outlined,and the application of gene modification of stem cells in ophthalmology are reviewed in this article.

Objective To investigate the methyenetetrahydrofolate (MTHFR) gene and angiotensin convertion enzyme (ACE) gene polymorphisms in pregnancy induced hypertension(PIH). Methods The MTHFR and ACE gene genotypes were determined in 62 pregnancy induced hypertension patients and 120 normal pregnant women by PCR RFLP. Results The frequencies of T allele(0.52) and the T/T genotype(27%) of MTHFR gene in PIH group were markedly higher than those in the control group(39% and 15% P

Objective:To identify pathogenicity of the potential splicing variants in two Chinese Han patients with osteogenesis imperfecta.Methods:Genomic DNA was extracted using the conventional phenol-chloroform method; whole exome sequencing (WES) was used to analysis the disease-related variants in the two probands; Minigene assay was used to identify pathogenicity of the variants found in the patients' genome that possibly affect RNA splicing.Results:Two potential splicing variants, c.858+1_858+5delGTAAG in intron 12 of COL1A1 and c.1405-7C>T in intron 24 of COL1A2, were found in proband 1 and proband 2, respectively. In addition, a missense mutation, c.2972G>T (p.G991V) in exon 45 of COL1A2, was detected in proband 2. Minigene assay revealed that the variant in proband 1 caused the skipping of exon 12, while the variant in proband 2 did not lead to aberrant splicing. G199 of the COL1A2 in proband 2 was a highly conserved amino acid site, and the results suggested that c.2972G>T (p.G991V) may be the real pathogenic variant by the means of bioinformatics analysis.Conclusion:The variant c.858+1_858+5delGTAAG in COL1A1 was a causative variant that led to OI in proband 1, while the missense variant c.2972G>T (p.G991V) in COL1A2 was the cause of OI in proband 2, instead of the variant c.1405-7C>T. Minigene assay for potential splicing variants detected by WES could not only validate the pathogenicity of the candidate variants and enrich the mutation spectrum of OI, but also lay the foundation for patients' prenatal diagnosis and subsequent mechanism research.

Objective To evaluate desmopressin stimulated inferior petrosal sinus sampling in diagnosing Cushing′s disease.Methods Sixteen ACTH-dependent Cushing′s disease patients underwent bilateral desmopressin stimulated inferior petrosal sinus ( IPS ) sampling because of negative or equivocal magnetic resonance imaging.Cortisol response to high-dose dexamethasone suppression test was also evaluated.ACTH sampling was taken from a peripheral vein and bilateral IPS before and both 5 and 10 min after injection of desmopressin.Diagnosis was based on the ratio of ACTH level in between IPS to peripheral vein by desmopressin test.Diagnosis was confirmed after surgery.Results High-dose dexamethasone suppression test showed suppressible in 9 of 16 patients with Cushing′s disease.An IPS gradient ＞2 was found in 14 of the 16 cases (87.5％ )with Cushing′s disease after desmopressin injection,while before injection the respective figure was 12 of 16 (75.0％).No severe adverse effects were observed during or after the procedure.Conclusion Desmopressin test during bilateral IPS sampling is a safe and effective diagnostic procedure in Cushing′s disease.

Objective:Iron accumulation is related to the occurrence of postmenopausal osteoporosis. Meanwhile, autophagy abnormality of bone marrow hematopoietic cells is observed in hip osteoporotic fracture. This study is performed to investigate correlation between iron accumulation induced bone loss and hematopoietic autophagy dysfunction to explore the new risk factor of osteoporosis.Methods:Male iron accumulation mice model was established by intraperitoneally injecting ferric ammonium citrate. Serum ferritin and osteogenic indicator P1NP were tested by ELISA. Bone mineral density was measured by micro-CT. Femur and tibia bone marrows were collected for hematopoietic stem and progenitor cells proportion and cell apoptosis analysis. Autolysosome formation was measured by image flow cytometry. We used conditional mouse model Atg7 flox/flox; Vav-Cre(Atg7 -/-) in which Atg7 had been genetically deleted in the hematopoietic system. Bone marrow hematopoietic stem and progenitor cells were collected for RNA sequence. micro-CT scan was conducted for Atg7 -/- femur. Results:Ferritin level of iron accumulation mice was significantly higher than control group( P<0.05). Iron accumulation inhibited P1NP and induced decreased bone mineral density( P<0.05). Iron accumulation bone marrow displayed enhanced hematopoietic stem and progenitor cells proportion( P<0.05), with more cell apoptosis( P<0.05). Hematopoietic autophagy was deteriorated in iron accumulation bone marrow. Transcriptomic profiling showed up-regulation of iron activity in Atg7 -/- mice, with increased iron homeostasis and iron membrane transporter genes, including Lcn2, Tfr2, Slc40a1(Fpn1), Steap3, and Cpox. micro-CT revealed severe bone loss and decreased bone mineral density in Atg7 -/- mice( P<0.05). Conclusion:Iron accumulation induced bone loss is related to inhibition of hematopoietic cells. Hematopoietic autophagy dysfunction is associated with bone loss.

Objective To explore relationship between the nature of carotid atherosclerotic plaque and the number and function of endothelial progenitor cells in peripheral blood.Methods A prospective study of 80 carotid atherosclerotic plaque patients were selected from June 2016 to March 2017 in Guangdong Second Provincial General Hospital.All patients were examined with Cranial magnetic resonance imaging or X-ray computed tomography,pathological examination,carotid artery color Doppler ultrasonography.Patients were divided into hard plaque group (n =42) and soft plaque group (n =38) according to the nature of carotid atherosclerotic plaque.Forty healthy subjects were selected as controls.Monocytes were obtained from 10 ml of elbow venous blood by density gradient centrifugation.Adherent cells were cultured and identified by confocal laser microscopy.The number,migration,proliferation and adhesion of EPCs in soft plaque group and hard plaque group were evaluated.Results The number of proliferating cells (0.847 ± 0.037),migrating cells(27.697±8.248) and adherent cells (46.184± 7.876) in the normal control group were significantly higher than those in the hard plaque group ((0.647±0.019),(18.643±3.289),(32.165±4.325)) and the soft plaque group ((0.679± 0.023),(23.576± 6.327),(40.587±6.523)) (all P＜ 0.001),while the proliferation,migration and adherent cells in the hard plaque group were lower than those in the soft plaque group (all P＜0.001).Conclusion The nature of carotid atherosclerosis plaque is closely related to the number and function of endothelial progenitor cells in peripheral blood.The number of endothelial progenitor cells in carotid atherosclerosis patients with hard plaque is small,and their proliferation,migration and adhesion ability are impaired.

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.
Humans ; Fetus ; Chloroform ; Computational Biology ; Ethnicity ; Ribs

Objective To explore the impact of macrophage-to-myofibroblast transition(MMT)on pulmonary fibro-sis induced by acute lung injury by LPS.Methods Totally 21 male mice were randomly classified into 7 groups:control group,model group(LPS-PF)at different time points and intervention group of clodronate-liposomes(CL-LIP)treatement at different time points(n=3).Pulmonary fibrosis was identified by HE and Masson staining microscopy.The immuno-fluorescence technology was used for the evaluation of numbers of macrophage-to-myofi-broblast transition cells(MMT cell which co-expressed CD68 and α-SMA).Bone marrow-derived macrophages(BMDMs)were randomly classified into two group:control(Ctrl)group and TGF-β1-treated group induced by transforming growthfactor-β1.α-SMA,FN and Col1 were detected by RT-qPCR.The expression of α-SMA,Smad3 and p-Smad3 protein was evaluated by Western blot.Results At day 7,the Ashcroft score of lung tissue in LPS-PF mouse model was significantly increased when compared with the Ctrl group(P＜0.01);While the score signifi-cantly declined when the model was pretreated with CL-LIP(P＜0.05).As detected by immuno-fluorescence stai-ning,in CL-LIP group the number of CD68-positive cells co-labeled with α-SMA was obviously less then that of LPS-PF group of the corresponding time point(P＜0.01).When the BMDMs were stimulated by TGF-β1 at 24 h,48 h and 96 h respectively,a higher expression of α-SMA,FN,Col1,were found in TGF-β1-treated group than that in Ctrl group at the corresponding time point(P＜0.01).The expression of Smad3,p-Smad3 significantly higher in LPS-PF group(at both day 7 and day 10)and TGF-β1-treated group(at both 48 h and 96 h)as compared to cor-responding control group(P＜0.01).Conclusions MMT promotes pulmonary fibrosis induced by ALI via LPS.Smad3 is proved to be involved in the MMT process.

Objective:To explore the predictive value of prognostic nutritional index(PNI)for recipients of heart transplantation.Methods:From January 2015 to December 2020, the relevant clinical data were reviewed retrospectively for 438 post-heart transplantation recipients at Union Hospital of Tongji Medical College of Huazhong University of Science and Technology.The optimal cut-off value of PNI was determined by receiver operating characteristic(ROC)curve.The inter-group differences of clinicopathologic characteristics were compared.Overall survival(OS)was calculated by Kaplan-Meier survival curve and compared by Log-rank test between two groups.The risk factors affecting OS were examined by univariate and multivariate Cox regression.Results:The optimal cut-off value of PNI was 49.3.They were assigned into two groups of low PNI(PNI≤49.3, 284 cases)and high PNI(PNI>49.3, 154 cases). As compared with high PNI group, low PNI group was significantly correlated with lower BMI, hemoglobin, neutrophil count, albumin, low-density lipoprotein, triglyceride, recipient/donor BMI ratio and recipient age and total bilirubin(all P<0.05). Cox regression multivariate analyses indicated that recipient age, waiting time, preoperative use of intra-aortic ballon pump(IABP)and lower PNI score were risk factors(all P<0.05). Kaplan-Meier survival curve revealed that OS was better in high PNI group than that in low PNI group( χ2=15.122, P<0.01). The area under the ROC curve of PNI for predicting OS was 0.603. Conclusions:PNI score upon admission is correlated positively with postoperative survival of heart transplantation patients.PNI has some predictive value for OS.When PNI score upon admission≤49.3, postoperative survival rate is relatively worse with a shorter postoperative survival.

The actor network theory, with reference to systems science, sociology and management science, offers a new perspective for research of the technology-society binary view. This theory as introduced by the authors, analyzed the constituent bodies and their relationship of the operation of the precise medical service system, and studied the translation process of the actor network. The analysis showed that the main factors affecting the operation of the precision medical service system were hospitals, patients, governments, scientific research institutions, technology-based enterprises, and universities, which were suppliers, demanders, managers, funders, and technical supporters of precision medical services. Among them, 82.72%(335/405) of clinicians believed that hospitals were the core actor that affected the operation of precision medical service system. 71.60%(290/405) of the clinicians thought that the training of medical workers was the current focus of the hospital, i. e., the mandatory access point for this actor network. Through training, the accurate medical service ability of medical workers and hospitals can be improved.