A decade of liver organoids: Advances in disease modeling

Liver organoids are three-dimensional cellular tissue models in which cells interact to form unique structures in culture. During the past 10 years, liver organoids with various cellular compositions, structural features, and functional properties have been described. Methods to create these advanced human cell models range from simple tissue culture techniques to complex bioengineering approaches. Liver organoid culture platforms have been used in various research fields, from modeling liver diseases to regenerative therapy. This review discusses how liver organoids are used to model disease, including hereditary liver diseases, primary liver cancer, viral hepatitis, and nonalcoholic fatty liver disease. Specifically, we focus on studies that used either of two widely adopted approaches: differentiation from pluripotent stem cells or epithelial organoids cultured from patient tissues. These approaches have enabled the generation of advanced human liver models and, more importantly, the establishment of patient-tailored models for evaluating disease phenotypes and therapeutic responses at the individual level.

HIV-1 subtypes and drug-resistance mutation sites in HIV/AIDS patients with antiretroviral-therapy failure / 上海预防医学

ObjectiveTo analyze the characteristics of HIV-1 subtypes and drug-resistance mutation sites among HIV-infected patients who received high-efficiency antiretroviral therapy but failed. MethodsA total of 130 plasma samples were collected from the patients who received antiviral treatment for 6 months in Taizhou City of Zhejiang Province in 2021 but failed the treatment and the viral load was ≥1 000 copies·mL-1. Nucleic acid in the samples was extracted， and the pol gene was amplified by nested reverse transcription PCR. After next-generation sequencing， online tools were used to compare and analyze the subtypes and drug-resistant mutation sites. ResultsA total of 110 samples were successfully sequenced. The main HIV-1 subtype was CRF01_AE， accounting for 42.72% （47 cases）， followed by CRF07_BC， 35.45% （39 cases）； CRF08_BC， 10.00% （11 cases）； CRF85_BC ， 8.18% （9）； and a small number of B subtype， 1.81% （2 cases） and C subtype， 1.81% （2 cases）. The online tool comparison showed that there were 67 cases with mutations of drug-resistance sites and 61 cases with drug-resistance. The mutation sites were mainly M184V， K103N， K65R and V181C， and the mutation rates were 20.00% （22 cases）， 10.91% （12 cases）， 8.18% （9 cases） and 8.18% （9 cases）， respectively. These mutation sites caused different degrees of resistance to nucleoside reverse transcriptase inhibitors （NRTI）， non- nucleoside reverse transcriptase inhibitors （NNRTI） and protease inhibitors （PI）， including 45 cases of NRTI， 61 cases of NNRTI and 2 cases of PI resistance. ConclusionThe HIV infected people who fail the treatment in Taizhou are mainly with the subtypes CRF01_AE and CRF07_BC. The rate of drug-resistance mutation is at a moderate level， mainly due to the mutation of NRTI and NNRTI drug-resistance sites， and a small number of PI drug-resistance sites. Therefore， the antiviral treatment plan for HIV infected people should be reasonably adjusted， and the detection of drug-resistance mutation sites should be strengthened to avoid the generation of transmissible drug-resistance strains.

Phenotypes and ATP7B gene variants in 316 children with Wilson disease / 中华儿科杂志

Objectives: To summarize the clinical phenotypes and the variation spectrum of ATP7B gene in Chinese children with Wilson's disease (WD) and to investigate their significance for early diagnosis. Methods: Retrospective analysis was performed on the clinical data of 316 children diagnosed as WD in Guangzhou Women and Children's Medical Center during the period from January 2010 to June 2021. The general situations, clinical manifestations, lab test results, imaging examinations, and ATP7B gene variant characteristics were collected. The patients were divided into asymptomatic WD group and symptomatic WD group based on the presence or absence of clinical symptoms at the time that WD diagnosis was made. The χ2 test, t test or Mann-Whitney U test were used to compare the differences between groups. Results: Among the 316 children with WD, 199 were males and 117 were females, with the age of 5.4 (4.0, 7.6) years at diagnosis; 261 cases (82.6%) were asymptomatic with the age of 4.9 (3.9, 6.4) years; whereas 55 cases (17.4%) were symptomatic with the age of 9.6 (7.3, 12.0) years. The main symptoms invloved liver, kidney, nervous system, or skin damage. Of all the patients, 95.9% (303/316) had abnormal liver function at diagnosis; 98.1% (310/316) had the serum ceruloplasmin lever lower than 200 mg/L; 97.7% (302/309) had 24-hour urine copper content exceeding 40 μg; only 7.4% (23/310) had positive corneal K-F rings, 8.2% (23/281) had abnormal MRI signals in the lenticular nucleus, and all of them had symptoms of damage in liver, kidney or nervous system. Compared with the group of symptomatic WD, asymptomatic group had higher levels of serum alanine aminotransferase and lower levels ceruloplasmin and 24-hour urine copper [(208±137) vs. (72±78) U/L, (55±47) vs. (69±48) mg/L, 103 (72, 153) vs. 492 (230, 1 432) μg; t=9.98, -1.98, Z=-4.89, all P<0.001]. Among the 314 patients completing genetic sequencing, a total of 107 mutations in ATP7B gene were detected, of which 10 are novel variants, and 3 cases (1.0%) had large heterozygous deletion (exons 10 to exon 11) in ATP7B gene. The percentage of missense mutation in asymptomatic WD children was significantly higher than that in symptomatic WD (81.5% (422/518) vs. 69.1% (76/110), χ²=8.47, P<0.05). WD patients carrying homozygous variant of c.2 333G>T had significantly low levels of ceruloplasmin than those not carrying this variant ((23±5) vs. (61±48) mg/L, t=-2.34, P<0.001). Conclusions: The elevation of serum ALT is an important clue for early diagnosis of WD in children, while serum ceruloplasmin and 24-hour urine copper content are specific markers for early diagnosis of WD. In order to confirm the diagnosis of WD, it is necessary to combine the Sanger sequencing with multiplex ligation-dependent probe amplification or other testing technologies.

Etiological analysis of acute respiratory tract infection in Taizhou of Zhejiang Province from 2020 to 2021 / 上海预防医学

ObjectiveWe examined the principal respiratory pathogens in patients with acute respiratory tract infection in Taizhou， Zhejiang Province during 2020‒2021 to provide evidence for prevention， diagnosis and treatment of acute respiratory tract infection. MethodsFrom September 2020 to August 2021， a total of 2 831 cases with acute respiratory tract infection were collected from two influenza sentinel surveillance hospitals in Taizhou， which had then received the examination of 22 respiratory pathogens by multiple fluorescence quantitative PCR. ResultsThe total positive rate of respiratory pathogens in 2 831 samples was 14.13%， among which enterovirus （7.77%） and respiratory syncytial virus （1.59%） were the principal pathogens. Except enterovirus， there was no significant difference in the positive rate of pathogens detected by gender（P>0.05）. Moreover， there was significant difference in pathogens by age （P<0.05）， with the highest positive rate in 0‒4 years（35.21%）. There was also significant difference in pathogens by seasons （P<0.05）， with the highest positive rate in summer（20.54%）. ConclusionThe positive rate of acute respiratory tract infection decreases significantly， compared with that before the COVID-19 epidemic. The differences in the positive rate differ significantly by age and seasons. Comprehensive consideration of diverse factors before diagnosis and the utilization of multiple fluorescent quantitative PCR can quickly and effectively determine the pathogens in the early stage of infection. Our findings may provide certain support for the diagnosis and treatment of acute respiratory infections in the context of COVID-19 in Taizhou.

Genetic and Clinical Characteristics of A Family with Combined PROC and PROS1 Genetic Variants / 中国实验血液学杂志

OBJECTIVE@#To test the anticoagulation functions, perform the genetic diagnosis and analyze the clinical characteristics in a family with combined heterozygous genetic variants of PROC and PROS1.@*METHODS@#Peripheral blood was collected from all the family members. Hematological phenotypes and activity of anticoagulant factors were analyzed. Target genes were amplified by PCR from DNA isolated from peripheral blood, and then were analyzed by Sanger DNA sequencing.@*RESULTS@#Many members in the family displayed the combined genetic variants in protein C and protein S, and six family members accompanied by deep venous thrombosis (DVT). The influences of genetic and secondary factors on the incidence of venous thrombosis in the family members were analyzed. The results showed that in this family, carriers of combined protein C and protein S gene defects had a higher incidence of VTE, but acquired factors still played a key role in the eventual thrombotic symptoms.@*CONCLUSION@#Venous thromboembolism (VTE) is a multifactorial disease, the combined genetic heterozygous mutations of protein C and S is an important genetic factor, and the clinical phenotype show a high heterogenicity, the secondary factors contribute to the VTE incidence.

Effects of mild hypothermia on the expression of high mobility group protein B1 in lung tissues of septic mice / 中国综合临床

Objective:To investigate the effects of naturally occurring mild hypothermia and artificial mild hypothermia on the expression of high mobility group box 1(HMGB1) in lung tissues of septic mice.Methods:One hundred and twenty BALB/C mice (SPF level) were randomly numbered.Twelve mice with integer multiples of 10 were used as the normal control (NC) group, and the remaining 108 mice were chosen as the septic group.The septic mouse model was established by intra abdominal injection of lipopolysaccharide (LPS) 10 mg/kg.The NC group was given the same dose of normal saline.Anal temperature of the septic mice were measured 1 hour after the model was established successfully, and then were divided into naturally occurring mild hypothermia group and non-mild hypothermia group according to T≤36℃ and T>36℃.In the naturally occurring mild hypothermia group, the mice with T<34℃ were eliminated, and the remaining septic mice were randomly divided into the naturally occurring mild hypothermia(NOMH) observation group and the keep normothermia (KN) group.NOMH group was not given preheating intervention, while KN group was placed in an incubator to maintain the anal temperature between 36.0℃ and 37.5℃.Septic mice in the non-mild hypothermia group were randomly divided into the nonhypothermia (NH) observation group and the artificial mild hypothermia (ATMH) group.The NH group was not treated with hypothermia, while the ATMH group was treated with physical hypothermia, so that the anal temperature of the mice were maintained at 34℃-36℃.Four mice in each group were randomly selected at 6 and 12 hours after modeling, and the concentrations of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and HMGB1 in serum were detected by enzyme-linked immunosorbent assay(ELISA). At 12 hours, the survival rate of each group of mice was observed.Then 4 mice of each group were sacrificed and lung tissues were taken.The pathological changes of lung tissues were observed by hematoxylin-eosin (HE) staining, and the expression of HMGB1 in lung tissues was observed by immunohistochemical staining.Real time fluorescence quantitative PCR and Western blot were used to detect the relative expression of HMGB1 at mRNA and protein levels.Results:(1)Twelve hours after modeling, the survival number of NOMH group, ATMH group, KN group and NH group were 36(40), 6(11), 27(40), 4(11), respectively, and there were differences between the four groups (χ 2=32.286, P=0.002). Compared with the other three groups of septic mice, the survival rate was highest in the NOMH group (compared with ATMH group: χ 2=5.222, P=0.022; compared with the KN group: χ 2=6.050, P=0.013; and the NH group: χ 2=11.672, P=0.001), but the differences between the other two groups were not statistically significant (all P>0.05). (2)Compared with the NC group, the concentrations of serum TNF-α, IL-6 and HMGB1 of septic mice in each group were significantly increased at 6 h and 12 h (all P<0.05). Compared with NOMH group, the concentrations of TNF-α, IL-6 and HMGB1 in ATMH group, KN group and NH group were significantly increased at 6 h and 12 h(all P<0.05), and the concentrations of TNF-α, IL-6 and HMGB1 in NH group were the highest at all time points (all P<0.05). The concentrations of TNF-α at 12 h decreased compared with 6 h (all P<0.05), while the concentrations of IL-6 and HMGB1 at 12 h increased compared with 6 h (all P<0.05). (3)HE staining showed that the lung tissue damage were minimal in NOMH group, followed by ATMH group.(4)Immunohistochemical staining showed that the expression of HMGB1 protein was in order of NOMH group, ATMH group, KN group and NH group; (5)The relative expressions of HMGB1 protein in lung tissues of septic mice in NOMH group, ATMH group, KN group, and NH group was 0.280±0.013, 0.320±0.016, 0.340±0.018, and 0.380±0.014, respectively, and the relative expression level of HMGB1 mRNA was 4.86±0.22, 6.02±0.18, 6.26±0.20, and 7.98±0.28, respectively, compared with NC group (HMGB1 protein content was 0.240±0.013, and the relative expression level of HMGB1 mRNA was 2.21±0.12) significantly increased (all P<0.05). Cmpared with NOMH group, the relative expression levels of HMGB1 protein and HMGB1 mRNA in the lung tissues of the ATMH group, KN group and NH group were significantly increased(all P<0.05), with the highest expression level in the NH group(all P<0.05). Conclusion:Mild hypothermia may reduce lung tissue damage by down-regulating the expression of HMGB1 in lung tissues of septic mice, and the improvement of spontaneous mild hypothermia was more significant.

Percutaneous Sacroplasty for Painful Sacral Metastases Involving Multiple Sacral Vertebral Bodies: Initial Experience with an Interpedicular Approach

OBJECTIVE: To report our initial experience of percutaneous sacroplasty (PSP) with an interpedicular approach for treating painful sacral metastases involving multiple sacral vertebral bodies. MATERIALS AND METHODS: This study prospectively enrolled 10 consecutive patients (six men and four women; mean age, 56.3 ± 13.8 years) who underwent PSP for painful sacral metastases involving multiple sacral vertebral bodies from March 2017 to September 2018. Visual analogue scale (VAS) scores, Oswestry disability index (ODI) values, and the number of opioids prescribed to the patients were assessed before and after PSP. The procedure duration, length of hospitalization, and complications were also recorded. RESULTS: Mean VAS and ODI declined significantly from 6.90 ± 1.20 and 74.40 ± 5.48 before the procedure to 2.70 ± 1.34 and 29.60 ± 14.57 after the procedure, respectively (p < 0.01). The median number of opioids prescribed per patient decreased from 2 (interquartile range [IQR] 1-3) pre-procedure to 1 (IQR 0–3) post-procedure (p < 0.01). Nine of the 10 patients showed no or decreased opioid usage, and only 1 patient showed unchanged usage. The mean procedure duration was 48.5 ± 3.0 minutes. The average length of hospitalization was 4.7 ± 1.7 days. Extraosseous cement leakage occurred in three cases without causing any clinical complications. CONCLUSION: PSP with an interpedicular approach is a safe and effective treatment in patients with painful sacral metastases involving multiple sacral vertebral bodies and can relieve pain and improve mobility.

Clinical application of a self-developed bone dust collector in mastoid cavity obliteration following mastoidectomy / 中华耳鼻咽喉头颈外科杂志

Objective@#To introduce a self-developed bone dust collector designed by the authors and evaluate its efficiency in mastoid obliteration following mastoidectomy.@*Methods@#Consecutive patients, from April 2017 to March 2018, who prepared to receive mastoidectomy were randomly divided into two groups, and in each group the bone dust was harvested by self-developed bone dust collector or by conventional used method respectively in mastoidectomy. The amount of the harvested bone dust and the time consumed in the collecting procedure were compared between two groups. The infection of the bone dust after mastoid obliteration was also evaluated during follow up.@*Results@#33 patients were recruited in bone dust collector group, and 31 patients in conventional method group.There is no significance of difference between two groups in sex ratio, age and pneumatization of mastoid cells (P>0.05 for all). The median amount of bone dust harvested by bone dust collector was significantly larger than that collected by conventional method (1.8 g vs 1.1 g, P<0.05). The median time spent in bone dust collector group was significantly shorter than that spent in conventional method group (4 minutes vs 6 minutes, P<0.05). No bone dust infection was found in the follow-up in all patients.@*Conclusion@#The present self-developed bone dust collector is a easy and useful apparatus which can significantly improve the efficiency of collecting bone dust in mastoidectomy.

Analysis on chemical constituents of fruit of Chaenomeles speciosa by UPLC-Q-Exactive Orbitrap-MS / 中草药

Objective An ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap-MS) method was developed to rapidly analyze and identify the chemical constituents from the fruit of Chaenomeles speciosa. Methods The analysis was performed on a Halo C18 column (100 mm × 2.1 mm, 2.7 μm) by gradient elution. The mobile phase consisted of 0.05% formic acid-acetonitrile and 0.05% formic acid-water at a flow rate of 0.3 mL/min. The column temperature was at 40 ℃. The information of the compounds was acquired in positive and negative mode. Results Totally 25 compounds were identified, including five kinds of free amino acids, 11 organic acids, two glycosides, three flavonoids, and four triterpenes. Conclusion The established method is accurate, rapid, and sensitive, which provides a reference for further clarifying the material basis of its efficacy and the selection of quality control indicators.

Genotyping and molecular tracing of the first local infection of dengue virus in Taizhou / 中华实验和临床病毒学杂志

Objective@#To determine the first dengue fever case in Taizhou and trace probable transmission sources.@*Methods@#Collected serum of three patients for antigen, antibody and nucleic acid detection. Dengue viruses were isolated and cultured in C6/36 cell. The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR) and then sequenced. The phylogenetic tree was drawn.@*Results@#Three cases were positive in nucleic acid detection. Two cases were IgM positive. One case was NSI antigen postive. Three strains of type I dengue virus were isolated from samples. The phylogenetic trees shown that the three strains were on the same branch. The identities of nucleotide were 99.87%. The identities of amino acid were 99.6%-99.8%.@*Conclusions@#The dengue virus strains isolated in Taizhou was imported from Guangdong or Southeast Asia and caused location infection.

Chronic subdural hematoma complicated with hepatic encephalopathy: a case report / 天津医药

@#This article report a case of patient with chronic subdural hematoma. After receiving a surgery, the patient presented mental symptoms such as abnormal behaviors and personality changes. The patient was diagnosed as hepatic encephalopathy and secondary epilepsy according to his medical history, laboratory tests, head CT / MRI and electroencephalogram. The secondary epilepsy was diagnosed by convulsion. After receiving corresponding therapy, these symptom disappeared.

Comparison of four score modes in prognosis assessment of AECOPD patients with respiratory failure / 实用医学杂志

Objective To explore the predictive effect of modified DECAF,DECAF,CAPS and APACHEⅡin the assessment of prognosis of AECOPD patients with respiratory failure. Methods Clinical data of 186 AECOPD cases complicated with respiratory failure were analyzed and four score modes were used within 24 hours of admission. Clinical endpoints were patients′ survival status 28 days after admission. The discriminative power of the four score modes was evaluated by the area under the receiver operating characteristic curve (AUC). Results AUC of the modified DECAF(0.777,95%CI:0.710-0.835) and DECAF (0.766,95%CI:0.699-0.825) for prognosis was significantly greater than that of CAPS(0.699 ,95%:0.628-0.764) and APACHEⅡ(0.715,95%:0.645-0.779). Conclusion The modified DECAF and DECAF have predictive values on assessing the prognosis of AECOPD patients with respiratory failure ,which are simple and efficient.

NR6A1 promotes vascular smooth muscle cell apoptosis by up-regulating RIPK3 gene expression / 中国病理生理杂志

AIM:To determine the role of nuclear receptor subfamily 6,group A,member 1 (NR6A1) in vascular smooth muscle cell (VSMC) apoptosis.METHODS:NR6A1 protein was over-expressed in the VSMCs by infection of adenovirus.The effect of NR6A1 on the viability of VSMCs was measured by MTT assay.DAPI staining,TUNEL staining and caspase activity assay were conducted.DNA microarray was used to quickly screen the target genes of NR6A1.The effect of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) silencing on NR6A1-induced apoptosis of the VSMCs was further analyzed.RESULTS:Adenovirus-mediated over-expression of NR6A1 induced the apoptosis of VSMCs.The RIPK3 gene expression was up-regulated by NR6A1 over-expression in the VSMCs.NR6A1-induced VSMC apoptosis was inhibited by RIPK3 silencing.CONCLUSION:NR6A1 promotes VSMC apoptosis by up-regulating the RIPK3 gene expression.

Fluorescent Detection of p-Nitroaniline Based on an Anionic Pyrenyl Probe / 分析化学

p-Nitroaniline ( PNA) was one kind of highly toxic aromatic amine and becomes an environmental pollutant in recent years. Here we reported the construction of a fluorescent sensor based on an anionic pyrene, 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS), for the rapid and visual detection of PNA in aqueous media. The fluorescence quenching of HPTS caused by the presence of PNA was through non-covalent interactions. A good linear relationship between fluorescent intensity of HPTS at 512 nm was obtained in the range of 10-120 μmol/L. The detection limit (3σ) of this approach was 4. 6 μmol/L. The results showed that the method was suitable for rapid detection of PNA in real samples with good sensitivity, selectivity, anti-interference, low cost and easy operation.

Fluorescent Detection of p-Nitroaniline Based on an Anionic Pyrenyl Probe / 分析化学

p-Nitroaniline ( PNA) was one kind of highly toxic aromatic amine and becomes an environmental pollutant in recent years. Here we reported the construction of a fluorescent sensor based on an anionic pyrene, 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS), for the rapid and visual detection of PNA in aqueous media. The fluorescence quenching of HPTS caused by the presence of PNA was through non-covalent interactions. A good linear relationship between fluorescent intensity of HPTS at 512 nm was obtained in the range of 10-120 μmol/L. The detection limit (3σ) of this approach was 4. 6 μmol/L. The results showed that the method was suitable for rapid detection of PNA in real samples with good sensitivity, selectivity, anti-interference, low cost and easy operation.

NR6A1 promotes vascular smooth muscle cell apoptosis by up-regulating RIPK3 gene expression / 中国病理生理杂志

AIM:To determine the role of nuclear receptor subfamily 6,group A,member 1 (NR6A1) in vascular smooth muscle cell (VSMC) apoptosis.METHODS:NR6A1 protein was over-expressed in the VSMCs by infection of adenovirus.The effect of NR6A1 on the viability of VSMCs was measured by MTT assay.DAPI staining,TUNEL staining and caspase activity assay were conducted.DNA microarray was used to quickly screen the target genes of NR6A1.The effect of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) silencing on NR6A1-induced apoptosis of the VSMCs was further analyzed.RESULTS:Adenovirus-mediated over-expression of NR6A1 induced the apoptosis of VSMCs.The RIPK3 gene expression was up-regulated by NR6A1 over-expression in the VSMCs.NR6A1-induced VSMC apoptosis was inhibited by RIPK3 silencing.CONCLUSION:NR6A1 promotes VSMC apoptosis by up-regulating the RIPK3 gene expression.

Association Between Rectus Abdominis Denervation and Ventilation Dysfunction in Patients with Amyotrophic Lateral Sclerosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Spontaneous potentials in electromyography (EMG) of paraspinal muscles are associated with diaphragm denervation and, therefore, poor respiratory function in amyotrophic lateral sclerosis (ALS) is understandable. EMG changes in the rectus abdominis (RA) display an effect similar to those in paraspinal muscles with respect to the function of lower motor neurons in the thoracic spinal cord. The RA denervation was examined to determine its association with ventilation dysfunction in ALS.</p><p><b>METHODS</b>We collected the clinical data of 128 patients with sporadic ALS in Department of Neurology of Peking University Third Hospital from 2009 to 2013. EMG, Revised ALS Functional Rating Scale (ALSFRS-R) and forced vital capacity (FVC) were performed in all patients and the differences in the EMG changes in RA between those with and without FVC ≥ 80% were analysed.</p><p><b>RESULTS</b>The mean FVC value was 83.4% ± 17.1% (range: 45%-131%) of the predicted value. A total of 79 patients displayed FVC ≥80%, and 49 patients displayed FVC <80%. Compared with the patients displaying a normal FVC (60/79, 75.9%), spontaneous activity in RA was significantly different among those patients displaying an FVC <80% (47/49, 95.9%). In addition, spontaneous potentials in RA were more frequently detected in patients exhibiting dyspnea (32/33, 97.0%) than in patients without dyspnea (75/95, 78.9%).</p><p><b>CONCLUSION</b>Spontaneous potentials in RA are associated with ventilation dysfunction and dyspnea in ALS patients.</p>

Identification of a novel pathogenic mutation in PDHA1 gene for pyruvate dehydrogenase complex deficiency / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the molecular genetic mechanism and genetic diagnosis of pyruvate dehydrogenase complex deficiency (PHD), and to provide a basis for genetic counseling and prenatal genetic diagnosis of PHD.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) was performed to amplify the 11 exons and exon junction of the PDHA1 gene from a child who was diagnosed with PHD based on clinical characteristics and laboratory examination results. The PCR products were sequenced to determine the mutation. An analysis of amino acid conservation and prediction of protein secondary and tertiary structure were performed using bioinformatic approaches to identify the pathogenicity of the novel mutation.</p><p><b>RESULTS</b>One novel duplication mutation, c.1111_1158dup48bp, was found in the exon 11 of the PDHA1 gene of the patient. No c.1111_1158dup48bp mutation was detected in the sequencing results from 50 normal controls. The results of protein secondary and tertiary structure prediction showed that the novel mutation c.1111 _1158dup48bp led to the duplication of 16 amino acids residues, serine371 to phenylalanine386, which induced a substantial change in protein secondary and tertiary structure. The conformational change was not detected in the normal controls.</p><p><b>CONCLUSIONS</b>The novel duplication mutation c.1111_1158dup48bp in the PDHA1 gene is not due to gene polymorphisms but a possible novel pathogenic mutation for PHD.</p>

Gene mutations and clinical manifestations in children with glycogen storage disease type Ib / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Glycogen storage disease type Ib (GSDIb) is caused by a deficiency of glucose-6-phosphate translocase (G6PT) activity due to SLC37A4 gene mutations. Most GSDIb patients have recurrent infections and inflammatory bowel disease, with poor prognosis. Detection of SLC37A4 gene mutations is of great significance for the diagnosis, subtyping and outcome prediction of GSD patients. This study aims to analyze SLC37A4 gene mutations in Chinese GSDIb patients and to investigate the relationship between its genotypes and clinical manifestations.</p><p><b>METHODS</b>All exons and their flanking introns of SLC37A4 gene in 28 Chinese children with a primary diagnosis of GSDIb were screened by PCR combined with direct DNA sequencing to detect SLC37A4 gene mutations.</p><p><b>RESULTS</b>Five SLC37A4 gene mutations were detected in 7 (25%) of the 28 children, i.e., p.Gly149Glu (9/13, 69%), p.Gly115Arg (1/13, 8%), p.Pro191Leu (1/13, 8%), c.959-960 insT (1/13, 8%) and c.870+5G>A (1/13, 8%).</p><p><b>CONCLUSIONS</b>In this study, c.959-960 insT is a novel mutation and p.Gly149Glu is the most common mutation. p.Gly149Glu may be associated with severe infections in children with GSDIb.</p>

The investigation on clinical features of acute pancreatitis / 中国医师进修杂志

Objective To explore the causes and clinical features change of acute pancreatitis,to get early diagnosis of severe acute pancreatitis.Methods The clinical data of 1195 cases of patients with acute pancreatitis were retrospectively analyzed.Results Main causes of acute pancreatitis were still biliary disease 31.72％ (379/1195),engorgement 25.44％ (304/1195) and hyperlipidemia 16.82％ (201/1195).But the causes of severe acute pancreatitis had changed.Hyperlipidemia was 44.59％ (140/314),biliary disease accounted for 28.34％ (89/314),engorgement accounted for 17.52％ (55/314).The incidence of hyperlipidemia severe acute pancreatitis was 69.65％ (140/201),the incidence of biliary severe acute pancreatitis was 69.88％ (58/83).The incidence of severe acute pancreatitis on patients with body mass index (BMI) ≤25 kg/m2,BMI26-30 kg/m2,BMI≥31 kg/m2 were 5.41％ (17/314),24.84％ (78/314),69.75％ (219/314).There was significant difference (P ＜ 0.05).The incidence of severe acute pancreatitis on patients at age ≤54 years-old,55-69 years-old,≥70 years-old were 12.42％ (39/314),21.66％(68/314),65.92％ (207/314).There was significant difference (P ＜ 0.05).Conclusions In shanghai,the main causes of acute pancreatitis still is biliary disease and engorgement in the lastest ten years.The causes of severe acute pancreatitis have changed.Hyperlipidemia is main casuse; the second one is biliary disease.The incidence rate of severe acute pancreatitis as well as high BMI and elder age were very high and serious.Thus control of blood-lipid and weight should be strengthened.