Objective:To carry out cytogenetic and molecular genetic analysis for two infertile patients carrying rare small supernumerary marker chromosomes (sSMC).Methods:Two infertile patients who received reproductive and genetic counseling at CITIC Xiangya Reproductive and Genetic Hospital on October 31, 2018 and May 10, 2021, respectively were selected as the study subjects. The origin of sSMCs was determined by conventional G banding, fluorescence in situ hybridization (FISH) and copy number variation sequencing (CNV-seq). Microdissection combined with high-throughput whole genome sequencing (MicroSeq) was carried out to determine the fragment size and genomic information of their sSMCs. Results:For patient 1, G-banded karyotyping and FISH revealed that he has a karyotype of mos47, XY, del(16)(p10p12), + mar[65]/46, XY, del(16)(p10p12)[6]/48, XY, del(16)(p10p12), + 2mar[3].ish mar(Tel 16p-, Tel 16q-, CEP 16-, WCP 16+ ). CNV analysis has yielded a result of arr[GRCh37]16p12.1p11.2(24999364_33597595)×1[0.25]. MicroSeq revealed that his sSMC has contained the region of chromosome 16 between 24979733 and 34023115 (GRCh37). For patient 2, karyotyping and reverse FISH revealed that she has a karyotype of mos 47, XX, + mar[37]/46, XX[23].rev ish CEN5, and CNV analysis has yielded a result of seq[GRCh37]dup(5)(p12q11.2)chr5: g(45120001_56000000)dup[0.8]. MicroSeq results revealed that her sSMC has contained the region of chromosome 5 between 45132364 and 55967870(GRCh37). After genetic counseling, both couples had opted in vitro fertilization (IVF) treatment and preimplantation genetic testing (PGT). Conclusion:For individuals harboring sSMCs, it is vital to delineate the origin and structural characteristics of the sSMCs for their genetic counseling and reproductive guidance. Preimplantation genetic testing after microdissection combined with high-throughput whole genome sequencing (MicroSeq-PGT) can provide an alternative treatment for carrier couples with a high genetic risk.

Collagen is a macromolecular protein constituting the extracellular matrix of animal connective tissue, which has been widely used and developed in fields of biomedicine, tissue engineering, food, and cosmetics. Due to its advantages such as abundant sources and good biocompatibility, low immunogenicity, and degradability, collagen can be used as a dressing or tissue engineering scaffold for wound repair. According to the source of materials, collagen can be divided into natural collagen and recombinant collagen. Natural collagen is mainly extracted directly from mammals and fish; recombinant collagen is obtained based on genetic engineering technology, and its sources include recombinant expression systems of microorganisms, animals, and plants. This paper summarizes the sources of collagen, and the roles, advantages, and disadvantages of different sources of collagen in wound repair, the particularity and superiority of collagen combined with three-dimensional printing technology in wound repair, the impact of market norms of China's collagen industry on the field of wound repair, and explains the precautions for the development of collagen-related products, aiming to provide new ideas for selecting a suitable source of collagen for wound repair.
Animals ; Collagen/metabolism* ; Wound Healing ; Tissue Engineering ; Tissue Scaffolds ; Cosmetics ; Mammals/metabolism*

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

With the rapid development of computing methods and computing power, the ability of artificial intelligence in problem modelling and analysis has been improved rapidly. Artificial intelligence has been gradually introduced into the field of medical care and nursing, and played an important role in assisting medical care and nursing. This paper mainly discusses the progress of artificial intelligence technology in nursing, and analyzes the existing problems and challenges, in order to provide new ideas for artificial intelligence in supporting the development of nursing more effectively.

Objective:To explore the effect of remote ischemic post-conditioning (RIPoC) on oxidation/reduction response, energy metabolism and inlfammatory reaction of ischemic myocardial tissue in rats with ischemic reperfusion (IR) injury.
Methods:IR model was established by 30 min left anterior descending (LAD) artery occlusion followed by 120 min reperfusion, conditioning was defined as 3 cycles of 30 seconds ischemia followed by 30 seconds reperfusion in adult rats. The rats were divided into 5 groups:①ischemic pre-conditioning (IPC) group, the rats received the conditioning prior to IR treatment,②ischemic post-conditioning (IPoC) group, the rats received 30 min LAD occlusion followed by conditioning at the beginning of 120 min reperfusion, ③ remote ischemic post-conditioning(RIPoC) group, the rats received 30 min LAD occlusion, followed by femoral artery conditioning at the beginning of 120 min reperfusion, ④ IR group, ⑤ Sham group. n=8 in each group.Ischemic myocardial tissue was collected at the end of experiment, superoxide dismutase (SOD) activity was assayed by xanthine oxidase method,malondialdehyde (MDA) content was examined by thiobarbituric acid method, myeloperoxidase (MPO) activity was determined by chemistry colorimetric method, adenosine triphosphate(ATP)amount was measured by bioluminescence method;expressions of myocardial stromal cell derived factor-1 (SDF-1) and vascular endothelial growth factor (VEGF), mitochondrial function related genes Ndufa2, Ndufa4, Cox4il and Cox7a2 were evaluated by real time quantitative PCR.
Results:In IPC, IPoC and RIPoC groups, the ischemic myocardial tissue had increased SOD activity and ATP amount, decreased MDA content and MPO activity; induced expressions of SDF-1, VEGF, mitochondrial function related genes Ndufa2, Ndufa4, Cox4il and Cox7a2.
Conclusion:RIPoC may increase anti-oxidation/reduction response, protect energy metabolism and reduce inlfammatory reaction in ischemic myocardial tissue, the effect was similar to pre-conditioning and post-conditioningin rats with IR injury.

The formation of physique was influenced by many factors and was closely related to the disease, especially by the social and cultural factors. According to the characteristics of physique, physique conditioning was conducive to rehabilitation of the disease. It was also the internal evidence for individualized treatment of rehabilitation. Traditional Chinese medicine (TCM) rehabilitation advocated functional rehabilitation as the main treatment purpose. Attentions were paid to promoteqi circulation. The psychological characteristics of the rehabilitation subject were especially emphasized on, in order to improve the therapeutic effect of rehabilitation. There were many classifications of physical evaluations, which were widely used in a variety of clinical diseases rehabilitation. The pathological physique correction and adjustment cannot be ignored in rehabilitation. Therefore, the application of physical evaluation in the guidance of rehabilitation therapy can enrich the content of TCM rehabilitation evaluation. It further improved TCM physical evaluation system to meet the needs for clinical practice and TCM modernization.

Objective: This study aimed to discriminate between Eupatorii Herba and its adulterants in order to guarantee the quality and clinical curative effect of this medicinal material. Methods: Genomic DNA extracted from Eupatorii Herba was used as templates. The internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA was amplified. Sequence assembly and consensus sequence generation were performed by CodonCode Aligner. The intraspecific and interspecific genetic distances of Eupatorii Herba and its adulterants were computed by MEGA5 and the phylogenetic tree was constructed using the neighbor-joining (NJ) method. Results: The length of ITS2 sequence of Eupatorii Herba was 218 bp. The maximum intraspecific genetic distance (K2P distance) of Eupatorii Herba was 0.0092. The minimum interspecific genetic distance of Eupatorii Herba and its adulterants was 0.024. The NJ trees showed that the ITS2 sequence would be used to identify Eupatorii Herba and its adulterants. Con-clusion: ITS2 sequence was able to identify Eupatorii Herba and its adulterants correctly and it provided a new technique to ensure clinical safety in utilization of traditional Chinese medicines.

Objective To study the calcium metabolism in tacrolimus(FK506)induced rats nephrotoxicity and the preventive effect of calcium channel blocker.Methods Twenty-four Spragueinduced or FK506-induced nephropathy model.Blood creatinine,blood electrolytes,renal tissue histopathology(HE stain)and the change of ultrastructural organization in renal cells by transmission electron microscope were observed.Results The blood creatinine levels of both CsA and FK506 groups [(36.00±2.61)and(34.17±4.54)μmol/L] were significantly higher than those of the FK506+Dilgroup and control group(all P＜0.05).The blood calcium levels of both CsA and FK506 groups (2.00±0.04 and 2.05±0.04 mmol/L) were significantly lower than those of the FK506+Dil group and control group(all P＜0.05).The blood creatinine and calcium levels of FK506+Dil group were not significantly different with those of control group(P＞O.05).Histopathology examination showed cloudy swelling and vacuolization of the renal tubular epithelial cells and intra-cellular mitochondria swelling and vacuolization in the CsA and FK506 groups.However,the pathological changes of the FK506+Dil group were remarkably milder in comparison with the CsA and FK506 groups.Concluum channel blocker,Dil,could prevent the FK506-induced nephrotoxicity.

AIM: To optimize the random amplified polymorphic DNA-Polymerase chain reaction(RAPD-PCR) system of Bupleurum chinense DC by comprehensive scoring method. METHODS: The L_(16)(4~5) orthogonal diagram was applied to studying the effects of different concentrations of Taq polymerase,Mg~(2+),dNTPs,primer and DNA on Bupleurum chinense DC.RAPD-PCR results and to get a suitable PCR system. RESULTS: A suitable RAPD reaction system was developed,i.e.1?Taq polymerase reaction buffer,2.5 mmol/L Mg~(2+),1.5 U Taq DNA polymerase,0.15 mmol/L dNTPs,0.4 ?mol/L primer and 25 ng genomic DNA templates in total 25 ?L reaction solution. CONCLUSION: The results show that the comprehensive scoring method could be used successfully to optimize the RAPD-PCR system.