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Objective:To explore the application of comprehensive simulated delivery room skill training combined with improved objective teaching in obstetric nursing teaching.Methods:A total of 84 nurses who practiced in the Department of Obstetrics of Nanjing Hospital of Traditional Chinese Medicine from August 2019 to August 2020 were selected as the research objects, and were divided into control group and research group according to the sequence of admission of practice nurses, with 42 nurses in each group. The control group adopted traditional teaching methods, and the research group adopted comprehensive simulated delivery room skill training combined with improved target teaching mode. The teaching effect of the two groups of practice nurses was evaluated by theoretical assessment, skill operation assessment and teaching satisfaction rate. SPSS 22.0 was used for t test and chi-square test. Results:The total score of theory assessment, single choice, multiple choice, blank filling, noun explanation, short answer and case analysis of practical nursing students in the study group were better than those in the control group, with statistically significant differences ( P<0.05). The results of practical skills operation and comprehensive quality ability evaluation of practice nurses in the study group were better than those in the control group, and the difference was statistically significant ( P<0.05). The teaching satisfaction rate of practice nurses in the control group was 80.95%(34/42), and that in the research group was 97.62%(41/42), with statistically significant differences ( χ2=6.10, P=0.014). Conclusion:The skill training of comprehensive simulated delivery room combined with improved objective teaching can improve the theoretical knowledge, practical skills and comprehensive quality of obstetric practice nurses, and improve the teaching satisfaction rate and obstetric nursing teaching quality of practical nurses.
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AIM: To analyze the effect of conbercept combined with laser photocoagulation on patients with macular edema secondary to branch retinal vein occlusion.METHODS: According to the random number table method, 102 patients(102 eyes)with macular edema secondary to branch retinal vein occlusion who treated in our hospital from January 2019 to June 2021 were divided into observation group and control group. There were 51 patients(51 eyes)in each group. The control group was treated with laser photocoagulation and the observation group was treated with conbercept combined with laser photocoagulation. The macular foveal retinal thickness, best corrected visual acuity and incidence of complications were compared among the patients of two groups.RESULTS: After treatment at 3mo, the retinal thickness of macular fovea in the observation group was lower than that in the control group(P<0.05), the best corrected visual acuity in the observation group was higher than that in the control group(P<0.05). The incidence of complications was not different between the two groups(P>0.05).CONCLUSION:The effect of conbercept combined with laser photocoagulation for macular edema secondary to branch retinal vein occlusion is remarkable to reduce the retinal thickness of macular fovea and improve the vision of patients and the complications did not increase compared with retinal laser photocoagulation therapy alone, which were safe and effective.
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BACKGROUND@#To observe the effects of Chinese medicine (CM) Polygonum cuspidatum (PC) on adenosine 5'-monophosphate-activated protein kinase (AMPK), forkhead box O3α (FOXO3α), Toll-like receptor-4 (TLR4), NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and monocyte chemoattractant protein-1 (MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism.@*METHODS@#A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups (n=12 per group). A normal group (n=6) was used as the control. Rats in the normal and model groups were administered distilled water (10 mL•kg) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol (23.33 mg•kg), and 7.46, 3.73, or 1.87 g•kg•d PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 mRNA and protein levels in renal tissue or serum.@*RESULTS@#Compared with the normal group, the mRNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6th and 8th weeks (P<0.01 or P<0.05). The mRNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated (P<0.01 or P<0.05). Compared with the model group, at the 6th week, the mRNA transcription levels of AMPK in the high- and medium-dose groups, and protein expression levels of AMPKα1, pAMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and pAMPKα1 in the mediumdose PC group, and pAMPKα1 in the low-dose PC group were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium- and low-dose PC groups were down-regulated (P<0.01 or P<0.05). At the 8th week, the mRNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, pAMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription levels of TLR4 in the medium- and low-dose PC groups, NLRP3 in the high- and low-dose PC groups and MCP-1 in the medium- and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated (P<0.01 or P<0.05).@*CONCLUSION@#PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecular mechanism of PC to improve hyperuricemia-mediated immunoinflflammatory metabolic renal damage.
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Animals , Male , Rats , AMP-Activated Protein Kinases , Physiology , Chemokine CCL2 , Blood , Disease Models, Animal , Fallopia japonica , Forkhead Box Protein O3 , Physiology , Hyperuricemia , Kidney Diseases , Drug Therapy , Plant Extracts , Pharmacology , Rats, Sprague-Dawley , Signal Transduction , Uric AcidABSTRACT
Objective To explore CTA and MRA characteristics of persistent trigeminal artery (PTA)with vascular disease. Methods 54 patients diagnosed as PTA by CTA and MRA were collected retrospectively,whose classification,variation and associated vascular disease were analyzed.Results There were more females than males in 54 patients,whose average age was 59.72±16.32,and the incidence of left ones were higher than that of the right ones.Type Ⅰ of Weon classification was 16 cases (29.63%),type Ⅱ 7 cases (12.96%),typeⅢ 16 cases (29.63%),type Ⅳ 1 1 cases (20.37%),type Ⅴ 4 cases (7.41%).The lateral type of Salas classification was significantly higher than that in the middle type.There were 16 cases (29.63%)with aneurysm,in which 3 cases (18.75%)were rupture.3 cases were with moyamoya disease (5.56%).39 cases (72.22%)were with basilar artery dysplasia.There were 33 cases (61.11%)with the postembryonic cerebral artery.There were 29 cases with cerebral infarction (53.7%),4 cases with cerebral hemorrhage (7.4%),3 cases with subarachnoid hemorrhage (5.5 6%).Cerebral arteriosclerosis with different degrees of stenosis was showed in 1 7 cases (31.48%). 2 cases were accompanied with trigeminal neuralgia (3.7%),and only 1 case was with ocular nerve palsy (1.85%).Conclusion CTA and MRA could be used to display the anatomical features and radiographic classification of PTA ,and also clearly display and evaluate PTA associated with vascular disease.
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This study aimed to investigate the inhibitory effect of total C-21 steroidal glycoside (TCSG) from Baishouwu on the proliferation, invasion and apoptosis of human hepatoma HepG2 cells in vitro and the relevant molecular mechanism. The experiment was divded into control group, TCSG groups (25, 60, 150 mg·L⁻¹) and positive control cisplatin group (1.33 mg·L⁻¹). Human hepatocyte L-02 cells and hepatoma HepG2 cells were treated with different concentrations of TCSG. Then, the inhibitory effect of TCSG on the proliferation of HepG2 cells was detected by CCK-8 method. Cell cycle, cell apoptosis and mitochondrial membrane potential were detected by flow cytometry. The apoptotic morphology was observed by Hoechst 33258 staining. Cell migration and invasion abilities were analyzed by Transwell chamber model. The protein expressions of Bcl-2, Bax, caspase 3, cleaved caspase 3 and Cyt C (cytosolchondrial) were detected by Western blot. Compared with the control group, the proliferation of HepG2 cells was significantly inhibited after treatment with different concentrations of TCSG for 48 h in a dose-dependent manner(<0.01), but no obvious effect was observed on the proliferation of L-02 cells. After treatment with TCSG for 48 h, apoptotic morphology such as nuclear shrinkage, fragmentation and semilunar or circular was observed; migration and invasion abilities of cells were significantly decreased, cell cycle was blocked in the G₀/G₁ phase(<0.01), mitochondrial membrane potential was remarkably decreased(<0.01), and so did the ratio of apoptosis(<0.01).Western blot results showed that the protein expressions of Bax, caspase 3, cleaved caspase 3, and Cyt C were significantly up-regulated(<0.05, <0.01), while the Bcl-2 protein was significantly down-regulated(<0.05, <0.01). Furthermore, the ratio of Bax/Bcl-2 was increased (<0.01). The results suggested that TCSG could inhibit the proliferation and invasion of HepG2 cells, and induce the apoptosis of HepG2 cells. The potential mechanism may be related to the blocking of cell cycle and the regulation of the expressions of apoptosis-related proteins by activating mitochondrial pathway.
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Objective To study the toxicological mechanisms of the compatibility application of Sargassum pallidum and Glycyrrhiza uralensis on kidney in rats. Methods Rats were divided into control, Sargassum pallidum (S), Glycyrrhiza uralensis (G), and Sargassum pallidum-Glycyrrhiza uralensis extract (S-G) groups, which were respectively exposed (gavages) for 4 weeks. Then, the levels of blood urea nitrogen (BUN), serum creatinine (Scr), aldosterone, cortisol, and electrolytes in rat serum and pathological sections of kidney were detected. Six active contents of Glycyrrhiza uralensis in kidney of rats were detected by UPLC-TQ/MS method. The expression of HSD11B2 in kidney was detected by Western blotting. Results Compared with the control group, all biochemical indicators of S group had no obvious change. It was found that the level of aldosterone from G group and S-G group was significantly lower than that from control group (P < 0.05, 0.01). In contrast to the control group, S. pallidum-G. uralensis extract led to significantly increased concentration of cortisol, BUN, and Scr in serum (P < 0.05, 0.01). The level of K+ and Cl- in S-G group was significantly lower than that in control group (P < 0.05, 0.01). Pathological examination showed that the G group had mild inflammation infiltration, and a serious inflammatory response accompanied by protein tube was absolved in S-G group. Compared with the G. uralensis extract group, the combination of S. pallidum and G. uralensis significantly raised the concentration of glycyrrhetinic acid (GA) in kidney (P < 0.05).When compared to that of control group, there was an inhibited expression of HSD11B2 in the kidney of L group and S-G group. Moreover, the expression of HSD11B2 in S-G group was markedly higher than that in G group (P < 0.05). Conclusion The toxicity of S-G group was mainly result that increased accumulation of GA, and inhibited the expression of HSD11B2, which resulted the aldosterone-cortisol system disorders.
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Objective To apply a simple fixator to salivary gland scintigraphy to evaluate its effect on body position movement. Methods Totally 30 female patients complaining xerostomia were randomly and equally divided into a conventional scan group and a simple fixator group,who were injected with Technetium Tc-99m Pertechnetate intravenously. In the conventional scan group the patients had their necks fixed with the bracket and cushion,while in the other group the fixation was executed with the single fixator. SPECT imager was used for dynamic salivary gland scintigraphy, and two nuclear medicine physicians evaluated the maximum shift of the parotid gland along left-right(X)and head-foot(Y)axes in series of dynamic images. SPSS 16.0 software was used for data analysis. Results In the conventional scan group the maximum shift was(6.6±4.6)mm at X axis and(5.2±3.4)mm at Y axis;in the single fixator group the maximum shift was(3.2± 1.6)mm at X axis and(3.0±1.3)mm at Y axis.There were significant differences between the maximum shifts in the two groups (P=0.012,X axis;P=0.027,Y axis).Conclusion The single fixator assists in salivary gland scintigraphy,and alleviates the body position movement during dynamic acquisition and provides data support for functional parameter calculation and result determination.
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Metabolic syndrome has become a global epidemic that adversely affects human health. Both genetic and environmental factors contribute to the pathogenesis of metabolic disorders; however, the mechanisms that integrate these cues to regulate metabolic physiology and the development of metabolic disorders remain incompletely defined. Emerging evidence suggests that SWI/SNF chromatin-remodeling complexes are critical for directing metabolic reprogramming and adaptation in response to nutritional and other physiological signals. The ATP-dependent SWI/SNF chromatin-remodeling complexes comprise up to 11 subunits, among which the BAF60 subunit serves as a key link between the core complexes and specific transcriptional factors. The BAF60 subunit has three members, BAF60a, b, and c. The distinct tissue distribution patterns and regulatory mechanisms of BAF60 proteins confer each isoform with specialized functions in different metabolic cell types. In this review, we summarize the emerging roles and mechanisms of BAF60 proteins in the regulation of nutrient sensing and energy metabolism under physiological and disease conditions.
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Humans , Chromatin Assembly and Disassembly , DNA-Binding Proteins , Metabolism , Disease , Metabolism , Nutrients , Metabolism , Signal TransductionABSTRACT
Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ～ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08％.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.
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<p><b>OBJECTIVE</b>To explore the molecular mechanism of exocrine immune inflammatory injury of Sjögren's Syndrome and the intervention of Banxia Qinlian Decoction (BQD).</p><p><b>METHODS</b>Totally 18 female NOD mice were randomly divided into the model group, the positive drug group, and the BQD group, 6 in each group. Six female BALB/c mice were recruited as a blank control group. Mice in the blank control group and the model group were gavaged with deionized water at the daily dose of 0.1 mL/10 g body weight. Tripterygium Tablet was administered by gastrogavage to mice in the positive group at the daily dose of 10 mg/kg. BQD was administered by gastrogavage to mice in the BQD group at the daily dose of 60 g crude drugs/kg. After 12 weeks of medication, mice were sacrificed. Their eyeballs were excised and blood collected. Tissues of bilateral parotids and submandibular glands were kept. mRNA transcriptional levels of IL-17, IL-6, type 3 muscarinic acetylcholine receptors (M3R), aquaporin protein-5 (AQP5) were detected by RT-PCR. Expression levels of M3R and AQP5 protein were detected by Western blot. Protein expression levels of IL-17 and IL-6 were detected by ELISA.</p><p><b>RESULTS</b>Compared with the normal group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly up-regulated in the model group (P < 0.01). Compared with the model group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly down-regulated in the positive drug group and the BQD group with statistical difference (P < 0.01, P < 0.05). Compared with the BQD group, mRNA-transcriptional levels of IL-17, IL-6, and M3R, as well as M3R and AQP5 protein expression levels were significantly down-regulated in the positive drug group (all P < 0.01).</p><p><b>CONCLUSION</b>The molecular mechanism of BQD in inhibiting SS exocrine neurotoxic injury might be possibly related to regulating Th17/IL-17 immune inflammatory way.</p>
Subject(s)
Animals , Female , Mice , Aquaporin 5 , Metabolism , Disease Models, Animal , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Interleukin-17 , Metabolism , Interleukin-6 , Metabolism , Mice, Inbred BALB C , Mice, Inbred NOD , Sjogren's Syndrome , Drug Therapy , Allergy and Immunology , Submandibular Gland , Th17 Cells , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of compound qingqin liquid (CQL) on Toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in rats with urate nephropathy, and to explore its renal protection mechanism.</p><p><b>METHODS</b>Totally 55 SD rats were randomly divided into 5 groups, i.e., the normal control group (n =5), the model group (n =10), the positive drug group (n=10), and the high-, medium-, low-dose CQL groups (n=10) respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. Allopurinol was intragastrically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intragastrically administered at the daily dose of 3.77, 1.89, 0.94 g/kg to rats in the high-, medium-, and low-dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively. Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay.</p><p><b>RESULTS</b>At week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4 mRNA among the 3 CQL groups (P > 0.05) at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>CQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.</p>
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Animals , Male , Rats , Disease Models, Animal , Drugs, Chinese Herbal , Pharmacology , Kidney , Metabolism , Kidney Diseases , Drug Therapy , Metabolism , RNA, Messenger , Genetics , Rats, Sprague-Dawley , Toll-Like Receptor 2 , Genetics , Metabolism , Toll-Like Receptor 4 , Genetics , Metabolism , Uric AcidABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Compound Qingqin Liquid (CQL) on the expression level of angiotensin II (Ang II) and COX-2 mRNA transcription and protein expression in the renal tissue of rats with uric acid nephropathy.</p><p><b>METHODS</b>SD rats were randomly divided into the blank control group, the model group, the positive drug group, the high, moderate, and low dose CQL group according to number randomization principle. The model was established by gastrogavage of adenine, accompanied with yeast feeding. Distilled water was given by gastrogavage to rats in the blank control group and the model group. Allopurinol at the daily dose of 9.33 mg/kg was given by gastrogavage to rats of the positive control group. CQL at the daily dose of 3.77 g/kg, 1.89 g/kg, and 0.09 g/kg was respectively given by gastrogavage to rats in the high, moderate, and low dose CQL groups. All treatment lasted for 6 weeks. Rats were randomly divided at week 4 (3 in the blank control group, and 6 in the rest groups), and the rest rats were killed at week 6. The renal tissue was extracted. The expression level of Ang II and COX-2 mRNA transcription were detected by RT-PCR. The expression level of Ang II was detected by ELISA. The expression level of COX-2 protein was detected by Western blot and immunohistochemical assay.</p><p><b>RESULTS</b>Compared with the blank control group, except the mRNA expression of Ang II at week 4, the mRNA and protein expression of Ang II and COX-2 obviously increased at week 4 and 6 in the model group (P < 0.01, P < 0.05). The COX-2 protein expression at week 4 was obviously lower in the high and moderate dose CQL groups than in the model group and the low dose CQL group (P < 0.05); the average integral of optical density value was obviously lower in the positive control group than in the model group. Except the mRNA expression of Ang II in the high dose CQL group at week 6, the mRNA and protein expression of Ang II obviously decreased in the positive control group and each dose CQL group (P < 0.01, P < 0.05). Of them, the effects were better in the high and moderate dose CQL groups than in the positive control group and the low dose CQL group (P < 0.05, P < 0.01). Besides, the mRNA expression of COX-2, the average integral of optical density value were obviously lower in the positive control group and each dose CQL group than in the model group (P < 0.05). The protein expression of COX-2 was obviously lower in the high and moderate dose CQL groups than in the model group (P < 0.05). Of them, the mRNA expression of COX-2 was better in the moderate dose CQL group than in the positive control group (P < 0.05); the protein expression of COX-2 was better in the high dose CQL group than in the low dose CQL group (P < 0.05).</p><p><b>CONCLUSION</b>CQL was capable of lowering the expression level of Ang II, COX-2 mRNA transcription and protein expression, thus suppressing the inflammatory pathological injury of the renal tissue.</p>
Subject(s)
Animals , Male , Rats , Angiotensin II , Metabolism , Cyclooxygenase 2 , Genetics , Metabolism , Disease Models, Animal , Drugs, Chinese Herbal , Pharmacology , Kidney , Metabolism , Kidney Diseases , Drug Therapy , Metabolism , RNA, Messenger , Genetics , Random Allocation , Rats, Sprague-Dawley , Uric AcidABSTRACT
Objective To study the effect of Bupi-Qufeng decoction on total serum IgE (TIgE) and EOS counts of atopic dermatitis (AD).Methods A total of 101 cases were randomly recruited into a treatment group and a control group.During the treatment both groups were give Danggui-Bohe ointment for external use,and based on this,oral Bupi-Qufeng decoction was added in the treatment group and loratadine in the control group.TIgE level and EOS count were detected both before the treatment and four weeks after the treatment.Results TIgE in the treatment group decreased significantly after the treatment (t=8.0063,P＜0.001),the decreased value of which was larger than the control group (t=3.6434,P＜0.001).EOS count in the treatment group also obviously decreased after the treatment (t=3.0314,P＜0.01) ; the decreased value of which was also larger than the control group (t=3.3331,P＜0.01).Conclnsion Bupi-Qufeng decoction could decrease TIgE and EOS level of AD patient,while the therapeutic mechanism needed further research.
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<p><b>OBJECTIVE</b>To study the molecular genetic mechanism of mucopolysaccharidosis type IV A(MPS IV A), and reveal the relationship between the genotype and phenotype, and provide a basis for prenatal gene diagnosis in the future.</p><p><b>METHODS</b>A preliminary diagnosis was made by qualitative detection of urinary glycosaminoglycans of the suspected MPS IV A proband. Then, mutation detection was performed on the proband and her family members with PCR and direct sequencing of the PCR products. After a novel c.1567T to G mutation was detected, Xsp I restriction enzyme digestion and amplification refractory mutation system (ARMS) fast specific identification were established to analyze the sequences of exon 14 in GALNS gene, including 110 randomly selected healthy controls, the proband and other pedigree members. At the same time, bioinformatic approaches for protein secondary, tertiary structure prediction were applied to identify the novel pathologic mutation.</p><p><b>RESULTS</b>The proband's urine GAGs test was a weak positive(± ), and a c.1567T to G heterozygous termination codon mutation in exon 14 and a c.374C to T heterozygous missense mutation in exon 4 were found. The proband was compound heterozygous of the two mutations, so was her younger sister. Her mother was a carrier with only a c.1567T to G heterozygous mutation in exon 14. Her father had a heterozygous mutation of c.374C to T in exon 4. After Xsp I restriction enzyme digestion, healthy controls had three bands including 28 bp, 120 bp and 399 bp, while the proband and her mother had four bands consisting of 28 bp, 120 bp, 148 bp and 399 bp. For amplification by ARMS specific primers, it was negative for the controls, while it was positive for the proband and the carrier. The results of protein secondary and tertiary structure prediction showed that the c.1567T to G mutation located in the stop codon, resulted in stop codon (TAG) changing to glutamic acid (GAG), with the peptide chain extending 92 amino acid residues, and secondary and tertiary protein structure change, which were not found in the controls. The result of enzyme assay showed that the activity of GALNS enzyme in the affected child was 8.3 nmol/17h/mg pr, which was obviously lower than the normal value (the normal range is 41.9-92.1 nmol/17h/mg pr).</p><p><b>CONCLUSION</b>These results illustrate that the c.1567 T to G is a novel pathologic mutation, which is the main cause of the disease in this family.</p>
Subject(s)
Child , Female , Humans , Infant , Amino Acid Sequence , Asian People , Genetics , Base Sequence , Chondroitinsulfatases , Chemistry , Genetics , Metabolism , Genotype , Molecular Sequence Data , Mucopolysaccharidosis IV , Genetics , Mutation , Genetics , Pedigree , Protein Conformation , Sequence AlignmentABSTRACT
<p><b>OBJECTIVE</b>To identify the mutations of iduronate-2-sulfatase (IDS) gene, to reveal its mutation features, and to establish a basis for genetic counseling and prenatal gene diagnosis of Hunter syndrome.</p><p><b>METHODS</b>Urine glycosaminoglycans (GAGs) assay, PCR and DNA sequencing were performed to detect mutation of IDS gene of the patient and his parents.</p><p><b>RESULTS</b>The result showed that the patient was: DS(++), HS(++), KS(-), CS(-), and that both of his parents were negative. A frame-shift deletion mutation (1062 del 16) was identified in exon 7 of the patient's IDS gene. His parents' genotypes were normal.</p><p><b>CONCLUSION</b>The patient's mutation was not inherited by his parents but a novel one. The mutation probably altered the primary structure and tertiary structure of IDS enzyme protein remarkably and lowered the activity of IDS enzyme greatly. Therefore it is supposed to be the direct cause of the disorder.</p>