IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

Endometrial injury diseases are an important factors in female infertility, reduced menstruation, and adverse pregnancy outcomes. Clinical characteristics include intrauterine adhesions, a thin endometrium, and reduced endometrial receptivity. Restoring the morphology of the uterine cavity and promoting endometrial reconstruction are the main goals of treatment. At present, hormone medication and surgery are the major Western medicine treatments. Traditional Chinese Medicine has an advantage in treating endometrial injury and preventing postoperative recurrence. Traditional Chinese Medicine diagnosis focuses on "kidney deficiency" as the primary condition,"blood stasis" as the secondary condition, and combined with "liver depression". Therapeutic strategies aim to "replenish kidney""nourish blood""dispel blood stasis", and improve the endometrium. By means of combination of disease and syndrome, classical prescription addition and subtraction, clinical effective experience, combination of acupuncture and medicine and external treatment of Traditional Chinese Medicine can improve the effect and prognosis of patients with endometrial injury diseases. This article reviews the progress of combined Traditional Chinese and Western Medicine treatment of endometrial injury diseases to provide reference for its clinical treatment.

Endometrial injury diseases are an important factors in female infertility, reduced menstruation, and adverse pregnancy outcomes. Clinical characteristics include intrauterine adhesions, a thin endometrium, and reduced endometrial receptivity. Restoring the morphology of the uterine cavity and promoting endometrial reconstruction are the main goals of treatment. At present, hormone medication and surgery are the major Western medicine treatments. Traditional Chinese Medicine has an advantage in treating endometrial injury and preventing postoperative recurrence. Traditional Chinese Medicine diagnosis focuses on "kidney deficiency" as the primary condition,"blood stasis" as the secondary condition, and combined with "liver depression". Therapeutic strategies aim to "replenish kidney""nourish blood""dispel blood stasis", and improve the endometrium. By means of combination of disease and syndrome, classical prescription addition and subtraction, clinical effective experience, combination of acupuncture and medicine and external treatment of Traditional Chinese Medicine can improve the effect and prognosis of patients with endometrial injury diseases. This article reviews the progress of combined Traditional Chinese and Western Medicine treatment of endometrial injury diseases to provide reference for its clinical treatment.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

This study aims to obtain nanoantibody of vaccinia virus protein D8 by SUMO tag and analyze its binding activity with antigen.The sequence was designed and synthesized to construct the prokaryotic expression plasmid pKMD-SUMO-D8,which was transformed into Escherichia coli BL21(DE3)receptor cells and induced to express.Anti-SUMO-D8 with His and SUMO tags were purified,and then all tags were removed by SUMO protease to further obtain Anti-D8.Final-ly,the binding activity was detected and analyzed by indirect ELISA and IFA.The results showed that the prokaryotic expression plasmid pKMD-SUMO-D8 and recombinant strains were construc-ted.Under the condition of 30 ℃,the expression of Anti-SUMO-D8 was best when induced by IPTG with 0.1 mmol/L for 4 h.High purity Anti-SUMO-D8 and Anti-D8 nanoantibodies were ob-tained.Indirect ELISA results showed that both Anti-SUMO-D8 and Anti-D8 had binding activity with antigen.Under the same conditions,the binding activity of Anti-D8 to vaccinia virus and E8L was higher than that of Anti-SUMO-D8.Indirect IFA results showed that 48 h after infection with vaccinia virus,some nuclei of the experimental group showed green fluorescence under fluorescence microscope.Nanoantibodies of vaccinia virus protein D8 could be efficiently prepared by SUMO tag,and the prepared nanoantibodies had binding activity.This study provides new idea for the preparation of nanoantibody,and also lays a foundation for further study of the biological function of vaccinia virus protein D8 nano-antibody.

Objective:To construct a primary nanobody library for human papillomavirus 16(HPV16)L1 protein and obtain a nanobody specific to HPV16 L1 through selection and identification.Methods:HPV16 L1 protein was used as antigen to immunize alpaca,and a primary antibody library was constructed using phage display technology.After three rounds of screening,positive clones were identified by ELISA.The VHH sequence of the strongest positive clone was used for eukaryotic expression.The target nanobody was obtained after affinity purification,gel filtration chromatography,SDS PAGE and WB identification.The affinity between the nanobody and HPV16 L1 protein was evaluated using surface plasmon resonance(SPR)technology.The cytotoxicity of the nanobody was detected using CCK-8 assay.The neutralizing activity of nanobody against HPV16 pseudovirus was detected using a luciferase reporter gene assay.Results:The primary library was constructed with a capacity of 1.304×1010 and an abundance of 6.5×109 clones/mL.ELISA identified 36 positive clones.Protein monomer and dimers were expressed and purified,and the target nanobody(designated as"Nb")was successfully identified.The binding affinity of Nb to HPV16 L1 protein was 35.41 nmol/L.There was no significant difference in HaCat cell proliferation activity between Nb group and blank group(P>0.05).Compared to the negative group,both 0.1 and 1 μmol/L Nb inhibited pseudovirus infection in 293FT cells(all P<0.01).Conclusion:This study successfully obtained a nanobody with high purity and strong affinity that exhibited no cytotoxicity to epithelial cells and effectively inhibited HPV16 pseudovirus infection in 293FT cells.The nanobody provides a promising candidate antibody-based drug for the prevention and treatment of HPV 16 infection.

Objective:To explore the potential genetic causes of unexplained neonatal encephalopathy.Methods:This retrospective study enrolled 113 infants diagnosed with unexplained neonatal encephalopathy and underwent genetic testing in the Children's Hospital of Hunan Province from January 2019 to May 2021. Perinatal data, clinical manifestations, electroencephalograph, brain MRI findings, genetic information, and prognosis of those patients were analyzed. T-test or Chi-square test were used for data analysis. Results:Of the 113 infants enrolled, 74 (65.5%) were males. The gestational age at birth was (38.6±1.5) weeks, and the birth weight was (2 957±561) g. The most common clinical manifestation was the disturbance of consciousness (83/113, 73.5%), followed by seizures (39/113, 34.5%). There were 38.2% (34/89) of the patients with abnormal brain MRI, and 80.4% (74/92) presented abnormal electroencephalography. Among the 113 infants, 60 (53.1%) had genetic abnormalities, including 48 with single nucleotide variations, eight with copy number variations, and four with chromosome abnormalities. Single nucleotide variations in the 48 patients were classified into syndromic ( n=18, 37.5%), metabolic ( n=16, 33.3%), epileptic ( n=11, 22.9%) and mitochondrial-related genes ( n=3, 6.3%), of which 14 were not included in any database. Among the 103 cases which were successfully followed up until December 31, 2021, 75 (72.8%) had a poor prognosis, including 52 (50.5%) death cases and 23 (22.3%) cases of development retardation. Birth weight and the incidence of seizures in the poor prognosis group were both lower than those in the non-poor prognosis group [(2 876±536) vs (3 254±554) g, t=3.15; 29.3% (22/75) vs 53.6% (15/28), χ2=5.20; both P<0.05], while the incidence of disturbance of consciousness was higher [80.0% (60/75) vs 53.6% (15/28), χ2=7.19, P<0.05]. The proportion of infants with genetic abnormalities in the poor prognosis group was higher than that in the non-poor prognosis group, but the difference was not statistically significant [53.3% (40/75) vs 46.4% (13/28), χ2=0.39, P=0.533]. Conclusions:Genetic abnormality is one of the leading causes of unexplained neonatal encephalopathy. Nucleotide variation is the most common genetic type. Syndromic, metabolic, and epileptic variants are frequently detected in these patients.

Objective To explore the design of word-picture matching semantic judgment task based on Chinese high frequency nouns and further explore the way to apply the task in a functional magnetic resonance imaging (fMRI)/event-related potential (ERP) study.Methods The materials, stimuli and procedure of the word-picture matching semantic task were provided. Then, a healthy subject for the pilot study was included. In the pilot study, fMRI and ERP data of the subject were collected during the word-picture matching task. The fMRI and ERP data were analyzed to test the feasibility of the word-picture matching task in an fMRI/ERP task.Results The results of fMRI analysis showed an increased activity in the right middle frontal gyrus under the word-picture presenting condition compared with the "+" presenting condition. In addition, fMRI analysis showed an increased activity in the right middle temporal gyrus in the word-picture mismatching condition compared with the word-picture matching condition. The results of ERP analysis showed an increased activity in the left frontal/temporal area in the word-picture matching condition and an increased activity in the right frontal area in the word-picture mismatching condition.Conclusion Language associated brain regions can be identified in the fMRI/ERP research based on the word-picture matching task described in this article, which indicated that the task is effective in exploring language processing mechanism in the brain.