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Objective To explore the effects of growth hormone(GH)on the proliferation,cycle,inva-sion,and migration of colon cancer cells and its possible mechanism.Methods GH3 cells with growth hor-mone-type pituitary adenoma were cultured in vitro,and the secretion of growth hormone in the supernatant of GH3 cells was detected by ELISA.Colon cancer LoVo cells in logarithmic growth phase were randomly divid-ed into the control group and the experimental group.PBS was added to the control group,while high concen-trations of recombinant GH were added to the experimental group.The two groups of cells were cultured in vitro under the same conditions.CCK-8 method was used to detect the proliferation of the cells.Flow cytome-try was used to detect the cell cycle.Transwell assay was used to detect the effect of growth hormone on the invasion and migration of the cells.Western blot was used to detect the expressions levels of E-cadherin,N-cadherin,Vimentin,and Snail-1 proteins in the cells.Results The results of ELISA showed that GH3 cells could secrete a large amount of GH,and the concentration of GH in the supernatant was(1 208±9)ng/mL.GH promoted cell growth in a dose-dependent manner within a certain concentration range,and GH 200 ng/mL was the optimal intervention concentration for subsequent experiments.Compared with the control group,the cell cycle in the experimental group changed from G1 phase to S phase and G2 phase,the ratio of G1 phase cells decreased,and the ratio of S phase cells and G2 phase cells increased(P<0.05).Compared with the control group,the number of the cell invasion and migration increased in the experimental group(P<0.05),the expression levels of N-cadherin,Vimentin,and Snail-1 was up-regulated,while the expression level of E-cadherin was down-regulated(P<0.05).Conclusion High concentration of GH promotes the prolifera-tion,invasion and migration of colon cancer cells,and induces the transition of cell cycle from G1 to S and G2 phases.The mechanism may be related to the epithelial-mesenchymal transition(EMT)of colon cancer cells promoted by high concentration of GH.
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BACKGROUND:Previous studies have shown that human dental pulp stem cells have good osteogenic differentiation potential and are potential seed cells in bone tissue engineering,and the effect of recombinant human growth hormone on the proliferative osteogenic differentiation of human dental pulp stem cells is still unclear. OBJECTIVE:To explore the effect of recombinant human growth hormone on the proliferation and osteogenic differentiation of human dental pulp stem cells. METHODS:Human dental pulp stem cells were isolated and cultured by tissue block culture method.After screening according to the drug concentration gradient,recombinant human growth hormone containing 10,100,250,500,1 000 μg/L was selected as the experimental group,and 0 μg/L without recombinant human growth hormone was selected as the control group.CCK-8 detection reagents were used on days 1,3,5,and 7 after the drug intervention to detect the proliferation of human dental pulp stem cells.Different concentrations(10,100,250,500,and 1 000 μg/L)of recombinant human growth hormone were added to the osteogenesis induction solution to intervene in human dental pulp stem cells.Alkaline phosphatase activity was detected by alkaline phosphatase staining and semi-quantitative analysis on day 7 of mineralization induction.The mRNA expression levels of osteogenic gene type I collagen,osteocalcin and Runt-related transcription factor 2 were detected by fluorescence quantitative RT-qPCR.Alizarin red staining was performed on day 14 of mineralization induction to detect osteogenic mineralized nodules. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that from the third day of intervention,the 100,250,500,1 000 μg/L recombinant human growth hormone group could promote the proliferation of human dental pulp stem cells compared with the control group(P<0.01).(2)The alkaline phosphatase activity of human dental pulp stem cells in the 100,250,and 500 μg/L recombinant human growth hormone group was significantly increased compared with the control group(P<0.01).The number of alizarin-stained mineralized nodules in human dental pulp stem cells in the 100,250 μg/L recombinant human growth hormone group was significantly increased compared with the control group(P<0.01).Compared with the control group,the mRNA expression of type I collagen and osteocalcin increased in the 250 μg/L recombinant human growth hormone group(P<0.05,P<0.01).mRNA expression of Runt-associated transcription factor 2 increased in the 100 and 250 μg/L recombinant human growth hormone groups(P<0.01).(3)According to the above results,recombinant human growth hormone at a concentration of 250 μg/L is a more suitable concentration to promote the proliferation and osteogenic differentiation of human dental pulp stem cells.
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Generally, small for gestational age(SGA) infants will catch up with growth after birth, but some SGAs fail to show enough catch-up growth, leading to physical growth backwardness, and the risk of metabolic diseases in adult offspring increases. The application of exogenous growth hormone replacement therapy can ensure and promote the occurrence of SGA catching up with growth. However, as growth hormone exerts therapeutic effects in related clinical diseases, clinical attention is gradually being paid to whether growth hormone may bring long-term risks. This article aims to review the efficacy and potential risks of growth hormone treatment for SGA.
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With the changes in various factors such as genetics and the environment, the incidence of childhood precocious puberty has been gradually increasing. Improving height is one of the key issues in the clinical management of precocious puberty. Currently, gonadotropin-releasing hormone analogs (GnRHa) remain the preferred treatment for precocious puberty, but their effect on height improvement is influenced by multiple factors, which may result in lower-than-expected height benefits. Combining recombinant human growth hormone (rhGH) therapy with GnRHa treatment is an alternative strategy to enhance the efficacy of GnRHa, but there is still no clear recommendation regarding the timing of their combination. Considering the current status of precocious puberty treatment, it is crucial to reevaluate the effects of GnRHa monotherapy and combination therapy with rhGH on height improvement. This article discusses strategies such as combination therapy indications to guide clinical medication and help children with precocious puberty achieve optimal height benefits.
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Niño , Humanos , Pubertad Precoz/tratamiento farmacológico , Hormona de Crecimiento Humana , Terapia CombinadaRESUMEN
@#Objective To study the effect of growth hormone(GH)with progesterone-primed ovarian stimulation(PPOS)protocol on in vitro fertilization(IVF)outcomes among women with low prognosis.Methods This is a retrospective cohort study using propensity score matching(PSM)analysis.Women commencing their IVF between January2017 to December2021,with and without GH co-treatment,were reviewed.Results After PSM,76 pairs of women with low prognosis were included into analysis.Paired testing showed there is a statistical increase in transferable embryo(P<0.001)in the GH co-treatment group comparing with control group.No significant difference showed in retrieved oocyte number and metaphaseⅡ(MⅡ)oocyte rate,two-pronuclear(2PN)zygote rate on day1,high-quality embryo rate,or clinical pregnancy rate,neither in gonadotropin(Gn)requirement,duration or peak estradiol.Subgroup analysis results showed transferrable embryo rate on day3 rising among decreased ovarian reserve women(P=0.010).For women aged below 40,MⅡoocyte rate(P=0.010)and transferrable embryo rate on day3(P<0.001)increased in GH co-treatment group.Conclusions GH is suggested a beneficial impact on oocyte quality and transferrable embryos with PPOS protocol in IVF.Its adjuvant administration can be proposed as an optional therapeutic strategy in women with low prognosis.
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Abstract Objective To explore the relationship between Growth Hormone Insulin-like Growth Factors (GH-IGFs) and growth retardation in children with bronchial asthma. Methods 112 children with bronchial asthma and 50 healthy children were studied. Serum GH, IGF-1, and Insulin-like Growth Factor Binding Protein 3 (IGFBP3) were assessed by ELISA. GH-IGFs-related parameters were compared, and the correlation between the parameters and bronchial asthma severity was analyzed. The bronchial asthma group was divided into the growth retardation group and non-growth retardation group to analyze the diagnostic value of GH-IGFs in growth retardation and the relationship between GH-IGFs and growth retardation. Results GH, IGF-1, and IGFBP3 in the bronchial asthma group were lower. GH, IGF-1, and IGFBP3 levels were decreased with the severity of bronchial asthma. GH, IGF-1, and IGFBP3 in the growth retardation group were lower than those in the non-growth retardation group. The AUC of GH-IGFs combined detection was higher than that of GH and IGFBP3 alone detection. GH < 9.27 μg/L and IGF-1 < 179.53 mmoL/L were risk factors for growth retardation in patients with bronchial asthma. Conclusion GH-IGFs-related parameters have diagnostic value for growth retardation in children, and decreased levels of GH and IGF-1 are risk factors for growth retardation in children.
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ABSTRACT Objective: This research aimed to evaluate retrospectively the effect of anastrozole on height gain and sex hormone levels in pubertal boys receiving growth hormone (GH). Materials and methods: Pubertal boys who received both GH and anastrozole (GH+A) were one-to-one matched with boys who received only GH (GH-Only) for chronological and bone age, pubertal stage and height before the GH initiation, treatment duration and midparental height. Anthropometric measurements throughout treatment and adult heights were compared between the groups. Sex hormone levels were evaluated longitudinally in the GH+A group. Results: Forty-eight cases (24 in each group) were included. There was no statistical difference in adult height between the GH+A and GH-Only (p = 0.071). However, when the analysis was limited to those receiving anastrozole for at least 2 years, mean adult height was higher in the GH+A than in the GH-Only group (173.1 ± 6.2/169.8 ± 5.6 cm, p = 0.044). Despite similar growth rates between the two groups, bone age advancement was slower in the GH+A than in the GH-Only in a mean anastrozole treatment period of 1.59 years (1.37 ± 0.80/1.81 ± 0.98 years, p = 0.001). The greatest increase for FSH, LH, total and free testosterone and decrease for estradiol levels were observed in the third month after anastrozole was started, albeit remaining within the normal ranges according to the actual pubertal stages. Conclusions: Using anastrozole with GH for at least 2 years decelerates the bone age advancement resulting in adult height gain with no abnormality in sex hormone levels. These results suggest anastrozole can be used as an additional treatment to GH for further height gain in pubertal boys.
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ABSTRACT Objective: To assess the effect of recombinant growth hormone (rGH) on body composition and metabolic profile of prepubertal short children born small for gestational age (SGA) before and after 18 months of treatment. Methods: It is a clinical, non-randomized, and paired study. Children born SGA, with birth weight and/or length <-2 standard deviations (SD) for gestational age and sex, prepubertal, born at full term, of both genders, with the indication for treatment with rGH were included. The intervention was performed with biosynthetic rGH at doses ranging from 0.03 to 0.05 mg/kg/day, administered subcutaneously, once a day at bedtime. Total lean mass (LM) and total fat mass (FM) were carried out using dual-energy X-ray absorptiometry (DXA), and the metabolic profile was assessed for insulin, glycemia, IGF-1 levels and lipid profile. Results: Twelve patients (nine girls, 8.17±2.39 y) were evaluated; three patients dropped out of the study. There was an increase of LM adjusted for length (LMI) (p=0.008), LMI standard deviation score (SDS) adjusted for age and sex (p=0.007), and total LM (p<0.001). The percentage of body fat (BF%) and abdominal fat (AF) remained unaltered in relation to the beginning of treatment. Among the metabolic variables, blood glucose remained within normal levels, and there was a reduction in the number of participants with altered cholesterol (p=0.023). Conclusions: The effect of rGH treatment was higher on LM than in FM, with increased LM adjusted for length and standardized for age and sex. Glycemia remained within the normal limits, and there was a decreased number of children with total cholesterol above the recommended levels.
RESUMO Objetivo: Avaliar o efeito do hormônio de crescimento recombinante (rHC) na composição corporal e no perfil metabólico de crianças pré-púberes com baixa estatura, nascidas pequenas para a idade gestacional (PIG) antes e depois de 18 meses de tratamento. Métodos: Estudo clínico, não randomizado e pareado. Foram incluídas crianças nascidas PIG, com peso e/ou altura ao nascer <-2 desvios padrão (DP) para idade gestacional e sexo, pré-púberes, nascidas a termo, de ambos os sexos, com indicação de tratamento com rGH. A intervenção foi realizada com rGH biossintético com doses variando de 0,03 a 0,05 mg/kg/dia, administrado por via subcutânea, uma vez ao dia ao deitar-se. A massa magra total (LM) e a massa gorda total (MG) foram determinadas por meio de absorciometria de raios X de dupla energia (DXA), e o perfil metabólico foi avaliado com dosagens de insulina, glicemia, IGF-1 e perfil lipídico. Resultados: Doze pacientes (nove meninas, 8,17±2,39 anos) foram avaliados; três pacientes abandonaram o estudo. Houve aumento da LM ajustada para estatura (LMI) (p=0,008), LMI standard deviation scores (SDS) ajustada para idade e sexo (p=0,007) e LM total (p<0,001). O percentual de gordura corporal (GC%) e gordura abdominal (AF) permaneceu inalterado em relação ao início do tratamento. Entre as variáveis metabólicas, a glicemia manteve-se na normalidade, e houve redução do número de participantes com colesterol alterado (p=0,023). Conclusões: O efeito do tratamento com HCr foi maior na MM do que na MG, com o aumento da MM ajustada para altura e padronizada para idade e sexo. A glicemia permaneceu normal e houve redução do número de crianças com colesterol total acima do recomendado.
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As alterações cromossômicas estruturais no cromossomo 14 são incomuns e podem levar a um espectro variado de manifestações clínicas, incluindo hipotonia, atraso no desenvolvimento psicomotor, déficits cognitivos e dismorfismos faciais. O fenótipo específico é influenciado pela localização, extensão e pontos de interrupção da deleção. Este relato de caso tem como objetivo detalhar o fenótipo e genótipo de uma pré-escolar com deleção 14q24.1, além de documentar a resposta ao tratamento com hormônio do crescimento recombinante humano (rhGH). A paciente, uma prematura nascida com medidas adequadas para a idade gestacional e filha de pais não consanguíneos, apresentou desconforto respiratório e dificuldades de deglutição no período neonatal, necessitando de gastrostomia até o primeiro ano de vida. Entre o nascimento e os dois anos e seis meses, ela apresentou uma redução da velocidade de crescimento e baixa estatura desproporcional. Foram observados também inclinação palpebral superior bilateral, baixa inserção das orelhas, fissura palatina posterior, dentição irregular, micrognatia, retrognatia, escoliose, encurtamento do fêmur direito com marcha antálgica, agenesia do rim esquerdo e posicionamento pélvico do rim direito. Além disso, a paciente exibiu atraso no desenvolvimento neuropsicomotor. A análise genética revelou uma deleção no braço longo do cromossomo 14 de aproximadamente 231 Kb. Com o tratamento com rhGH, observou-se melhora na taxa de crescimento e na estatura final. A evolução clínica do caso indica que a administração de rhGH, associada ao acompanhamento clínico rigoroso e ao tratamento das comorbidades, pode contribuir para a melhoria dos parâmetros antropométricos.
Structural chromosomal changes on chromosome 14 are uncommon and can lead to a diverse spectrum of clinical manifestations, including hypotonia, delayed psychomotor development, cognitive deficits, and facial dysmorphisms. The specific phenotype is influenced by the location, extent, and breakpoints of the deletion. This case report aims to detail the phenotype and genotype of a preschooler with 14q24.1 deletion, in addition to documenting the response to treatment with recombinant human growth hormone (rhGH). The patient, a premature female, born with appropriate measurements for gestational age and daughter of non-consanguineous parents, presented respiratory discomfort and swallowing difficulties in the neonatal period, requiring gastrostomy until the first year of life. Between birth and two years and six months, she presented a reduction in growth speed and disproportionate short stature. Bilateral upper eyelid tilt, low ear insertion, posterior cleft palate, irregular dentition, micrognathia, retrognathia, scoliosis, shortening of the right femur with antalgic gait, agenesis of the left kidney and pelvic positioning of the right kidney were also observed. Furthermore, the patient exhibited delayed neuropsychomotor development. Genetic analysis revealed a deletion on the long arm of chromosome 14 of approximately 231 Kb. With rhGH treatment, an improvement in growth rate and final height was observed. The clinical evolution of the case indicates that the administration of rhGH, associated with strict clinical monitoring and treatment of comorbidities, can contribute to the improvement of anthropometric parameters.
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ABSTRACT Objective: Congenital hypopituitarism (CH) is a rare disease characterized by one or more hormone deficiencies of the pituitary gland. To date, many genes have been associated with CH. In this study, we identified the allelic variant spectrum of 11 causative genes in Turkish patients with CH. Materials and methods: This study included 47 patients [21 girls (44.6%) and 26 boys (55.4%)] from 45 families. To identify the genetic etiology, we screened 11 candidate genes associated with CH using next-generation sequencing. To confirm and detect the status of the specific familial variant in relatives, Sanger sequencing was also performed. Results: We identified 12 possible pathogenic variants in GHRHR, GH1, GLI2, PROP-1, POU1F1, and LHX4 in 11 patients (23.4%), of which six were novel variants: two in GHRHR, two in POU1F1, one in GLI2, and one in LHX4. In all patients, these variants were most frequently found in GLI2, followed by PROP-1 and GHRHR. Conclusion: Genetic causes were determined in only 23.4% of all patients with CH and 63% of molecularly diagnosed patients (7/11) from consanguineous families. Despite advances in genetics, we were unable to identify the genetic etiology of most patients with CH, suggesting the effect of unknown genes or environmental factors. More genetic studies are necessary to understand the etiology of CH.
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Introducción. Los pequeños para la edad gestacional (PEG) suelen tener una talla final 1 DE bajo la media. Se diferencian tres grupos según antropometría al nacimiento: de peso reducido (PRN), de longitud reducida (LRN) o ambos. Objetivos. Describir las características de los pacientes PEG atendidos en el Servicio de Endocrinología Pediátrica de un hospital de tercer nivel, y analizar la evolución de niños PEG sin crecimiento recuperador a los 4 años de edad, en tratamiento con hormona del crecimiento (GH), según su diagnóstico. Métodos. Estudio retrospectivo de pacientes PEG atendidos desde 2004 hasta 2021. Resultados. Se estudiaron 89 PEG; 44/89 iniciaron tratamiento con GH (11/44 PRN, 8/44 LRN y 25/44 ambos). La edad media al diagnóstico fue de 3,87 años; la talla media al inicio del tratamiento fue de -2,99 DE en los PEG diagnosticados por PRN, -2,85 DE en aquellos diagnosticados por LRN y -3,17 DE en los diagnosticados por bajo PRN y LRN. La talla final fue de -1,77, -1,52 y -1,23 DE, respectivamente, lo que supone una ganancia total de 1,22, 1,33 y 1,93 DE, respectivamente, alcanzando así su talla diana con una diferencia de 0,36 ± 0,08 DE. Conclusión. Menos de la mitad de los PEG derivados a la consulta precisaron tratamiento con GH, por no tener la edad de 4 años aún, o haber completado el crecimiento recuperador. Aquellos pacientes PEG según peso y longitud al nacimiento presentaron percentiles peores al diagnóstico y una mayor respuesta a GH.
Introduction. Small for gestational age (SGA) children usually have a final height of 1 SD below the mean. Three groups are established based on anthropometric characteristics at birth: low birth weight (LBW), short birth length (SBL), or both. Objectives. To describe the characteristics of SGA patients seen at the Department of Pediatric Endocrinology of a tertiary care hospital and to analyze the course of SGA children without catch-up growth at 4 years of age who were receiving treatment with growth hormone (GH), according to their diagnosis. Methods. Retrospective study of SGA patients seen between 2004 and 2021. Results. A total of 89 SGA children were studied; 44/89 started treatment with GH (11/44 LBW, 8/44 SBL, and 25/44 both). Their mean age at diagnosis was 3.87 years; their mean height at treatment initiation was -2.99 SD in SGA children diagnosed by LBW, -2.85 SD in those with SBL, and -3.17 SD in those with both LBW and SBL. Their final height was -1.77, -1.52, and -1.23 SD, respectively, with a total gain of 1.22, 1.33, and 1.93 SD, respectively, thus reaching their target height with a difference of 0.36 ± 0.08 SD. Conclusion. Less than half of SGA children referred to the clinic required treatment with GH because they were not yet 4 years old or had not completed their catch-up growth. SGA patients according to birth weight and length had worse percentiles at diagnosis and a greater response to GH.
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Humanos , Preescolar , Estatura , Hormona de Crecimiento Humana/uso terapéutico , Hormona del Crecimiento , Estudios Retrospectivos , Edad GestacionalRESUMEN
Abstract Objective The present study aimed to evaluate the effects of GH treatment on the body composition of children born with SGA. Methods This study is a systematic review of the literature. CINAHL, Embase; Medline/Pubmed, Scopus and Web of Science were searched from inception to March 2022. Results Four studies met the inclusion criteria, with an intervention time of 1 to 3 years, using doses from 0.03 to 0.07 mg/kg/day of GH. Bone densitometry by dual-energy X-ray absorptiometry (DXA) with whole-body scans was the most used method to assess body composition. Most studies (n= 3) had SGA children as a control group with the same characteristics as the case group; the mean age was similar between the groups (minimum of 5.1 ± 1.4 years and maximum of 6.7 ± 1 0.8 years) and all participants had an average height ≤ -3DP. The Lean Mass (LM) and Fat Mass (FM) outcomes of the studies were not presented in a standardized manner; thus, they cannot be compared. There was a significant increase in LM in the group treated with GH in relation to the pre-treatment period and in comparison, to the untreated control group. Three studies showed a significant decrease in FM at the end of the intervention period, and in two studies, this decrease occurred in the control group. Conclusions Despite the differences in the presentation of results and in the evaluation periods, the results of the studies showed that growth hormone favors the gain and maintenance of lean mass, and it also affects fat mass reduction and redistribution.
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Abstract Objective: The main growth hormone action is to promote linear growth increasing protein synthesis stimulation and osteoblastic activity. Peak bone mass extends from adolescence to 30 years of age. Several factors may influence this acquisition and prevent fracture risk in adulthood, such as genetic potential, GH, ethnicity, and lifestyle factors. This study aims to compare bone mass and osteometabolic profile of white and Afro-descendant Brazilian adolescents in the transition phase, who were treated with human recombinant growth hormone in childhood. Methods: The authors selected 38 adolescents from the Transition Outpatient Clinic of the University of São Paulo. Lumbar spine and total body bone mineral density (BMD) and bone mineral content (BMC), serum calcium, 25-OH-vitamin D and bone markers were analyzed at the rhGH withdrawal. Results: The mean age was 16.8 ± 1.6 years. There were 21 Afro-descendant and 17 whites. Thirty-four percent of the sample presented vitamin D insufficiency, 66% inadequate calcium intake and 44.7% physical inactivity. The Afro-descendants showed a lower lumbar spine and total body Z scores than those of the whites (p = 0.04 and p = 0.03, respectively), as well as their mean body weight (p = 0.03). There were no differences in the remaining osteometabolic parameters. Conclusion: As most adolescents had vitamin D insufficiency, low calcium intake, and physical inactivity, calcium, and cholecalciferol supplementation and lifestyle changes should be encouraged. The Brazilian Afro-descendant may be a vulnerable group for low bone mass, requiring
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Growth hormone deficiency (GHD) has become a serious healthcare burden, and presents a huge impact on the physical and mental health of patients. Here, we developed an actively separated microneedle patch (PAA/NaHCO3-Silk MN) based on silk protein for sustained release of recombinant human growth hormone (rhGH). Silk protein, as a friendly carrier material for proteins, could be constructed in mild full-water conditions and ensure the activity of rhGH. After manually pressing PAA/NaHCO3-Silk MN patch to skin for 1 min, active separation is achieved by absorbing the interstitial fluid (ISF) to trigger HCO3 ‒ in the active backing layer to produce carbon dioxide gas (CO2). In rats, the MN patch could maintain the sustained release of rhGH for more than 7 days, and produce similar effects as daily subcutaneous (S.C.) injections of rhGH in promoting height and weight with well tolerated. Moreover, the PAA/NaHCO3-Silk MN patch with the potential of painless self-administration, does not require cold chain transportation and storage possess great economic benefits. Overall, the PAA/NaHCO3-Silk MN patch can significantly improve patient compliance and increase the availability of drugs, meet current unmet clinical needs, improve clinical treatment effects of GHD patients.
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@#Objective To optimize the expression of recombinant human growth hormone-Fc(rhGH-Fc)fusion protein in CHO cells in order to obtain better glycosylation ratio and lower content of highmannose.Methods CHO cells expressing rhGH-Fc were cultured in a 7 L bioreactor.The glycosylation modifications of rhGH-Fc were adjusted by improving the composition of feeding media(using three commercial media:Gly-1:EX-CELL Glycosylation Adjust,Gly-2:SHEFF-CHO PLUS PG ACF and Gly-3:EfficientFeed C + AGT Supplement & GlycanTune C + Total Feed),and the glycosylation type and proportion of the target proteins were analyzed by mass spectrometry.Results The G0F(main glycosylation types:G0,G1 and G2;F:fucose)of Gly-1,Gly-2 and Gly-3 were 32.89%,58.66% and 33.28%,the G1F were 31.39%,18.03%and 34.90%,and the G2F were 31.39%,18.03% and 34.90%,respectively.Gly-1 and Gly-3 made the target protein contain less G0F while more G2F;Gly-3 feeding scheme-showed less high mannose modification than the other two schemes.Conclusion Gly-1 medium changed the glycosylation modification from G0F to G1F and G2F,while Gly-2 medium changed that from G2F and G1F to G0F.However,Gly-3 medium changed the glycosylation modification from G0F to G1F and G2F,and the contentof high mannose was less than 5%,which may have a better effect on modifying glycosylation type and proportion of the target protein.
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@#ObjectiveTo optimize the condition for anion exchange chromatography in purification process of recombinant human growth hormone-Fc(rhGH-Fc)fusion protein by Design of Experiment(DOE)so as to decrease the content of host cell protein(HCP)in bulk of protein.MethodsA complete factorial experimental design with four factors at two levels was established by the DOE in Minitab 19 software.The condition(flow rate,sample load,pH value of buffer and salt concentration of eluent)for anion exchange chromatography in purification process of rhGH-Fc was optimized by DOE to find out the operating space.ResultsThe experimental results were predicted accurately by the established DOE model.The sample load,pH value of buffer,salt concentration of eluent as well as the interaction of pH value of buffer and salt concentration of eluent showed significant influence on the HCP content in the harvest.The optimal sample load,flow rate as well as pH value and salt concentration of eluent were 15 mg/mL,120 cm/h,7.0 ～ 8.0 and 0.1 mol/L respectively.The HCP contents in eluents under the optimal condition were less than 400 ng/mg,which met the requirements for verification within the range of results in determined operating space.ConclusionThe condition for removal of HCP by anion exchange chromatography was successfully optimized by DOE,which provided a reference for further application of DOE in the biopharmaceutical field.
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@#Primary growth hormone (GH) resistance or growth hormone insensitivity syndrome, also called Laron syndrome, is a hereditary disease caused by mutations in the GH receptor or in the post-receptor signaling pathway. This disorder is characterized by postnatal growth failure resembling GH deficiency. Differentiating the two conditions is necessary. We present the cases of two siblings, a 16-year-old female and a 9-year-old male, born from a consanguineous union. Both had normal birth weights with subsequent severe short stature and delayed teeth eruption, with no features suggestive of any systemic illness. Serum insulin-like growth factor 1 (IGF1) and insulin-like growth factor binding protein 3 (IGFBP3) were both low. Suspecting GH deficiency, provocative testing with clonidine was done revealing peak growth hormone >40 ng/mL in both patients. In view of low IGF1 and IGFBP3 and high GH on stimulation, IGF1 generation test was done for both siblings, with values supporting the diagnosis of GH insensitivity or Laron syndrome.
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Síndrome de LaronRESUMEN
In this study, we established a novel bioassay to determine the activity of polyethylene glycolated recombinant human growth hormone (PEG-rhGH) using Nb2-11 cells. We performed experimental condition optimization and methodological verification, and then detected the relative potency of PEG-rhGH products using this method. We demonstrated that the bioactivity of PEG-rhGH in promoting Nb2-11 cell proliferation displays a dose-response relationship, which conformed to the four-parameter model. Using PEG-rhGH reference as a control, we analyzed the relative potency of six batches of PEG-rhGH products, as well as linearity, regression and parallelism of the obtained curves. The relative potency of six batches of PEG-rhGH products was 95% to 105%. These results implied that the new bioassay established may be employed in quality control of PEG-rhGH products.
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Objective:To provide molecular evidence for clinical diagnosis and genetic counseling by analyzing the clinical characteristics and identifying the pathogenic genes in a SPONASTRIME-type spondyloepimetaphyseal dysplasia(SEMDSP)family.Methods:Clinical data of the family members was collected and analyzed. Case 2 was identified as the proband for whole-exome sequencing and variant analysis. Suspected variants were validated across family numbers using Sanger sequencing.Results:The two affected individuals in this family, a brother and a sister, both presented as short stature. The initial diagnosis for the sister(case 1)was made at the age of 4 years and 2 months(height: 88.6 cm), and for the brother(case 2)at 4 years and 4 months(height: 81.6 cm). Both individuals exhibited distinctive facial features, including frontal bossing, midface hypoplasia with depressed nasal bridge, upturned nostrils, ocular hypertelorism, and epicanthus, thick hair, short lingual frenulum, stubby fingers and palms, and absence of scoliosis. The parents displayed normal phenotypes. Laboratory tests indicated growth hormone deficiency in both affected individuals. Imaging studies revealed significant bone age delay in case 2, while case 1 showed longitudinal striations at the distal radius but with bone age matching their actual age(5 years and 11 months). Despite receiving recombinant human growth hormone treatment, both patients had inadequate responses. Genetic testing identified compound heterozygous mutations in the TONSL gene shared by the two siblings. These mutations included a paternally inherited c. 1291-14_1291-11delCCTC and a maternally inherited c. 1909_1920delACGCTGCAGCAG. Notably, SEMDSP families have not been reported in China, and the c. 1909_1920delACGCTGCAGCAG mutation is a novel variant.Conclusion:Two patients were diagnosed as spondyloepimetaphyseal dysplasia, SPONASTRIME type, and the compound heterozygous variant was the genetic cause of this family.
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Objective:To investigate the efficacy and safety of polyethylene glycol-recombinant human growth hormone (PEG-rhGH) in the treatment of preadolescent growth hormone deficiency (GHD) and idiopathic short stature (ISS).Methods:Clinical data of children with preadolescent GHD or ISS diagnosed in Chengdu Women and Children′s Central Hospital from January 2014 to October 2022 were retrospectively analyzed. Among them, 36 children with GHD received (0.19±0.02) mg·kg -1·week -1 PEG-rhGH for treatment; and 21 children with ISS received (0.20±0.01) mg·kg -1·week -1 PEG-rhGH. The changes of height, weight, bone age, insulin-like growth factor-1 (IGF-1), thyroid function, fasting blood glucose and fasting insulin were observed in the two groups during treatment. Results:The height of children in GHD group was (107.56±8.38)cm and (111.68±7.94)cm 6 and 12 months after treatment, which was significantly higher than that before treatment ((101.62±8.83) cm, P<0.05). The height of children in ISS group was (108.69±12.59)cm and (114.66±11.47)cm 6 and 12 months after treatment, which was significantly higher than that before treatment ((103.40±12.66) cm, P<0.05). The height of the two groups was increased the most during 0-3 months of treatment ((3.15±0.99) cm and (2.91±0.73) cm, respectively). After 12 months of treatment, body mass index and IGF-1 were significantly higher than those before treatment ( P<0.05), and bone age and maturity were not significantly different ( P>0.05). In the GHD group, growth rate was negatively correlated with actual age, bone age, height, weight, IGF-1 and fasting insulin before treatment. In the ISS group, growth rate was negatively correlated with pre-treatment height standard deviation score (HtSDS). During treatment, hypothyroidism occurred in 2 cases (1 case in GHD group and 1 case in ISS group), and serum IGF-1 level increased in 9 cases (6 cases in GHD group and 3 cases in ISS group), there was no severe adverse reactions. Conclusion:PEG-rhGH treatment has good efficacy in treatment of GHD and ISS, and the children with GHD may have better curative effect than those with ISS. The children in both groups have the fastest growth rate within 3 months after treatment. Short-term use of PEG-rhGH does not increase the body mass index and promote bone maturity, and has no significant effect on the level of thyroid function, blood sugar and insulin, and has no serious adverse reactions.