Analysis of gene mutation type and frequency in children with thalassemia aged 0-18 years in Chengdu, Sichuan / 中华检验医学杂志

Objective:To analyze the type and frequency of thalassemia gene mutation in children aged 0 to 18 years in Chengdu.Methods:A total of 568 children from Chengdu, who were initially positive for thalassemia during screening from September 2018 to July 2021, were recruited. Among them, there were 308 males and 260 females. The type of mutation and distribution of α and β types of thalassemia in this cohort was analyzed utilizing PCR reverse dot blot.Results:Among the 568 children, 356 were genetically diagnosed as thalassemia, with a total positive rate of 62.68%. Among them, there were 140 cases of α-thalassemia with a positive rate of 24.65%, and 202 cases of β-thalassemia with a positive rate of 35.56%. There were 14 carriers of α-complex β-thalassemia gene, and the positive rate was 2.46%. Among these cases, the types of α-thalassemia gene mutation were mainly αα/--sea (79.29%), αα/-α3.7 (7.86%), and-α3.7/--sea (7.14%) genotypes, accounting for 94.29% of all types. In the 202 β-thalassemia patients, 199 heterozygous mutations were identified, mainly including cd17(A?T) (36.13%), cd41-42(-TCTT) (32.68%), IVS-2-654(C?T) (20.79%), and accounting for 88.61% of all types of gene mutation, and 3 compound heterozygous mutations were detected. α-complex β-thalassemia was detected in 14 patients, including cd41-42(-TCTT)/-α3.7, VS-2-654(C?T)/--sea, cd17(A?T)/-α3.7 and cd41-42(-TCTT)/--sea, which accounting for 57.14% of all types of gene mutation. Our results showed that there is no sex difference between α and β thalassemia in Chengdu area, whereas the prevalence of α combined with β thalassemia is higher in males ( P=0.003). Conclusions:The type of α-thalassemia mutation in Chengdu is mainly αα/--sea, whereas β-thalassemia with cd17 (A?T) mutations and α-complex β-thalassemia are more frequent in males. This study provides a reference for the formulation of prevention and control strategies for thalassemia in Chengdu.

Diagnosis of the accurate genotype of HKαα carriers in patients with thalassemia using multiplex ligation-dependent probe amplification combined with nested polymerase chain reaction / 中华医学杂志(英文版)

BACKGROUND@#Patients carrying the HongKongαα (HKαα) allele and -α/ααα could be misdiagnosed as -α/αα by the current conventional thalassemia detection methods, leading to inaccurate genetic counseling and an incorrect prenatal diagnosis. This study was aimed to accurately analyze the genotypes of HKαα carriers and -α/ααα.@*METHODS@#Samples were collected in our hospital from July 2017 to October 2019. Twenty-four common types of Chinese thalassemia were screened by gap-polymerase chain reaction (Gap-PCR) and reverse dot blot (RDB). Anti-4.2 multiplex-PCR was used to confirm carriers of the ααα duplication with -α deletion. Two-round nested PCR and multiplex ligation-dependent probe amplification (MLPA) were applied to accurately identify and confirm their genotypes. For data analysis, we used descriptive statistics and Fisher's exact tests.@*RESULTS@#Two thousand five hundred and forty-four cases were identified as thalassemia in 5488 peripheral blood samples. The results showed that α, β, and αβ compound thalassemia were identified in 1190 (46.78%), 1286 (50.55%), and 68 (2.67%) cases, respectively. A total of 227 samples from thalassemia patients were identified as -α/αα by Gap-PCR, and the genotypes of two samples were uncertain. There was a difference between Gap-PCR and combined groups (Gap-PCR combined with nested PCR and MLPA) in detecting HKαα (P < 0.05). Among the 229 patients, 20 patients were identified as HKαα carriers and one was identified as -α/ααα by two-round nested PCR and MLPA, including 15 patients with HKαα/αα, three with HKαα/αα and β-thalassemia coinheritance, one with HKαα/--, one with HKαα/-α and β-thalassemia coinheritance, and one with -α/ααα and β-thalassemia coinheritance.@*CONCLUSIONS@#ααα and HKαα genotypes of patients carrying -α need to be detected to reduce the misdiagnosis rate of patients carrying HKαα and -α3.7/ααα alleles. More accurate genetic counseling can be provided in the clinic using nested PCR combined with MLPA.

Effect of primary tooth root resorption on the isolation of dental pulp stem cells from primary teeth / 中国组织工程研究

BACKGROUND: Mesenchymal stem cells are derived from a variety of tissues, such as bone marrow, pulp, placenta, umbilical cord and adipose tissue. Mesenchymal stem cells from deciduous pulp have strong stemness and biological activity, no rejection, and strong immunoregulation, which are one of excellent cell sources for biotherapy. It is easy and suitable for large-scale production of mesenchymal stem cells from deciduous pulp, thereby laying a good foundation for the industrialization of dental pulp stem cells. OBJECTIVE: To investigate the effect of primary tooth root resorption on the isolation and expansion of dental pulp stem cells, in order to further determine the proper period for tooth extraction for pulp stem cell isolation. METHODS: Totally 173 primary teeth from 173 pupils aged 7-9 years were extracted for the isolation and expansion of dental pulp stem cells. Before tooth extraction, we took X-ray periapical film or orthopantomography of the primary teeth, in accordance with the World Health Organization (WHO) professional inspection standard. Root resorption in primary teeth could be divided into five kinds: root resorption 1/3, root resorption 1/2, root resorption 2/3, complete root resorption, and natural loss of primary teeth. Collected teeth after tooth extraction were placed into a medium within 7 seconds, and stored at in a refrigerator of 2-4 ℃. Then, the teeth were sent to the Oral Stem Cell Bank in Beijing within 24 hours by a professional cold-chain logistics for the isolation, expansion and preservation of dental pulp stem cells. Statistical analysis of the test results was performed. RESULTS AND CONCLUSION: For 32 primary teeth with root resorption 1/3, dental pulp stem cells were successfully extracted from 30 teeth, with a success rate of 94%, and ectopic eruption of permanent teeth was found in 12 cases, with an average eruption time of (2.19±0.18) months. For 35 primary teeth with root resorption 1/2, dental pulp stem cells were successfully extracted from 32 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 11 cases, with an average eruption time of (1.89±0.13) months. For 59 primary teeth with root resorption 2/3, dental pulp stem cells were successfully extracted from 54 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 8 cases, with an average eruption time of (1.42±0.12) months. For 37 primary teeth with complete root resorption (the bottom of the pulp was intact), dental pulp stem cells were successfully extracted from 34 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 2 cases, with an average eruption time of (1.03±0.15) months. For 10 naturally exfoliated primary teeth, dental pulp stem cells were not extracted, and ectopic eruption of permanent teeth was found in 4 cases, with an average eruption time of (0.65±0.23) months. To conclude, the primary teeth naturally exfoliated have no dental pulp with no stem cells; the success rate of extraction is relatively high in primary teeth that have mobility I-II, root resorption 2/3 or complete root resorption but with the complete bottom of the pulp. Moreover, it has no effect on permanent tooth eruption, and it is the best time for collection of primary teeth.

Research progress in long non-coding RNAs involved in invasion and metastasis of breast cancer / 基础医学与临床

Long non-coding RNAs(lncRNAs) defined as RNAs of more than 200 nucleotides in length don't en-code protein but are closely related to the development of breast cancer.Plenty of lncRNAs are dysregulated in breast cancer,and they can regulate the invasion and metastasis of breast cancer through various mechanisms such as transcription and post-transcriptional regulation which is greatly important to guide the treatment and prognosis of breast cancer.

Analysis of NF1 gene mutations among eleven sporadic patients with neurofibromatosis type 1 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic etiology for 11 sporadic patients with neurofibromatosis type 1.</p><p><b>METHODS</b>Chip targeting capture and high-throughput sequencing were employed to detect potential mutations of NF1 and NF2 genes among the 11 patients. The data was filtered through multiple mutational databases and in-house whole exome sequence database. Sanger sequencing was used for analysis of family members of the patients.</p><p><b>RESULTS</b>Eleven pathogenic variants were found among the 11 patients, which included two splicing mutations, one missense mutation, two nonsense mutations, and six frame-shifting mutations. None of the mutations was recorded by the public database or the in-house database generated from 1775 samples through whole exome sequencing. None of the unaffected parents carried the same mutation. Seven mutations were associated with neurofibromatosis type 1 previously, while the remaining four were discovered for the first time. Prenatal diagnosis of two high-risk pregnancies suggested that neither fetus has inherited the NF1 mutation from their affected parents.</p><p><b>CONCLUSION</b>Identification of causative mutations in patients with sporadic-type neurofibromatosis type 1 has provided a basis for genetic counseling. The four novel mutations have enriched the spectrum of NF1 gene mutations.</p>

Thermal tissue effects of the second-generation argon plasma coagulation on human stomach tissue :An ex vivo study / 西安交通大学学报(医学版)

Objective To explore the thermal effects of the second-generation argon plasma coagulation (VIO APC/APC2) on ex vivo human stomach tissue with different mode ,output power and application time .Methods The thermal effects of VIO APC were studied on fresh surgery-resected human stomach tissues .Different output powers were combined with mode "forced (F )","pulse , effect 1 (P1 )" and "pulse , effect 2 (P2 )";three application time ,the depth and maximum diameter of each injury were recorded .Results The depth of tissue injuries was correlated with output power (P<0 .001) ,energy (P=0 .008) and mode (P=0 .013) .The maximum diameter of injuries was correlated with output power (P<0 .001) ,application duration (P=0 .001) ,energy (P<0 .001) and mode (P=0 .001) .The incidence of harmful damage was only related to mode (P=0 .012) .The depth of tissue injuries was P1> F> P2 (P<0 .05) ,and the maximum diameter of injuries was P2> P1> F (P<0 .05) . Conclusion Different modes of VIO APC can create different thermal effects on the gastric tissues .Compared with F mode ,P1 mode creates deeper injuries while P2 mode creates larger but shallow injuries .All the three modes may cause injuries involving muscularis propria .

Expressions of CD49f splicing isoforms in different cell lines / 基础医学与临床

Objective To compare the expression of CD 49f splicing isoforms in different human stem cells and colon cancer cell lines , and construct lentiviral vectors overexpressing specific isoform .Methods The expression of CD49f isoforms in different cells was detected by RT-PCR and real-time PCR.The lentiviral vectors overexpressing CD49f isoforms were constructed by molecular cloning method .The overexpression of CD49f in colon cancer cell line HT29 was confirmed by FCM, Western blot and real-time PCR, the invasive ability was examined by transwell assay.Results CD49f splicing isoform B was highly expressed in H 9 human embryonic stem cell line , while iso-form A expressed in epithelial cells .Both isoform A and B were expressed in mesenchymal cells .CD49f isoforms A and B were also expressed in colorectal cancer cell lines , while HT29 and HCT116 showed higher expression of iso-form A than isoform B , HCT8 and LoVo showed higher expression of isoform B than isoform A .Overexpression of CD49f isoform B greatly increased the invasive ability of HT 29 cells, while isoform A showed no effect .Conclusions The expressions of CD49f splicing isoforms A and B in different types of cells are significantly different , which sug-gests that CD49f isoforms play different biological functions in cells .

Sonic hedgehog signaling enhanced the expression of histone demethylase, lysine-specific demethylase 8 in the head and neck squamous cell carcinoma cell line SCC-6 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine whether the sonic hedgehog (Shh) signaling could regulate the expression of histone demethylases in the head and neck squamous cell carcinoma(SCC).</p><p><b>METHODS</b>Human recombinant SHH-N protein or over-expression of the mutant 2 smoothened (M2-SMO) was applied to activate the Shh signaling in tongue squamous cell carcinoma cell line-SCC-6 in this study. Cyclopamine was used to block the Shh signaling in SCC-6. The real-time reverse transcription (RT)-PCR was used to detect the expression of histone demethylases at the mRNA level.</p><p><b>RESULTS</b>The data showed that activation of the Shh signaling up-regulated the expression of histone demethylase, lysine-specific demethylase 8 (KDM-8) at the mRNA level by human recombinant SHH-N protein (1.841 ∼ 3.591 fold compare with untreated group; P < 0.01), over-expression of the M2-SMO also increased the expression of KDM-8 (1.358 ∼ 3.013 fold compared with empty vector group; P < 0.05), and after the Shh signaling was blocked by Cyclopamine, the expression of KDM-8 was down regulated (decreased 25.6% ∼ 66.6% compared with control cells, P < 0.05).</p><p><b>CONCLUSIONS</b>Histone demethylase KDM-8 was downstream target gene of Shh signaling in head and neck squamous cell carcinoma cell line SCC-6, and its expression was positively regulated by the Shh signaling.</p>

Causes of death after hip fracture in senile patients / 中华创伤杂志(英文版)

Hip trauma has been a leading cause of death in senile patients for more than a centenary. Although the mortality decreased due to the advanced technique in medication, surgery and nursing, the increasing mortality should not be neglected in elders after orthopedic operation nowadays. Many factors are considered to influence the causes of death after trauma, such as age, gender, personal customs, comorbidities, types of fracture, timing of surgery, procedure, anesthesia, complications, medical treatment, activity of daily living, or even marriage status. This article reviews these causes from the aspects of patient's own factors, iatrogenic factors, medical treatment and other factors and provides some clues for further clinical application according to the recent foreign and domestic researches. According to the present research, it is essential for surgeons to perform a comprehensive estimation for patients suffering from hip trauma.

Postoperative mortality and morbidity in octogenarians and nonagenarians with hip fracture: an analysis of perioperative risk factors / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the role of high risk factors in octogenarians and nonagenarians with hip trauma, which may lead to excessive mortality and morbi- dity postoperatively.</p><p><b>METHODS</b>Fifty-four octogenarians and nonagenarians patients were enrolled in the study, receiving surgical repair of hip fracture in our hospital from January 2006 to January 2010. High risk factors were recorded preoperatively in detail. Complications and survival state were followed up by telephone for 2 years postoperatively. All the data were analyzed by Chi-square test with SPSS 13.0.</p><p><b>RESULTS</b>Twenty-six males (48.1%), aged from 80 to 94 years with a mean age of 84.2 years, and twenty-eight females (51.9%), aged from 80 to 95 years with a mean age of 83.4 years, were presented in the cohort study. The hip traumas were caused by daily slight injuries (52 cases) and car accidents (2 cases), respectively. Twenty-eight patients (51.9%) with femoral neck fracture while 26 patients (48.1%) with intertrochanteric fracture were diagnosed through an anterior-posterior pelvic radiophotograph. In this series, 39 patients (72.2%) suffered from one or more comorbidities preoperatively. The morbidity was 48.1% and the major cause was urinary tract infection, while a significant difference was noted between females and males. The mortality was 20.4% with a predominant cause of acute renal failure.</p><p><b>CONCLUSIONS</b>The gender should be considered as a critical high risk factor in octogenarians and nonagenarians with hip trauma postoperatively. Females are more likely to suffer complications postoperatively, which is especially obvious in senile patients over 80 years (P less than 0.05). Urinary tract infection is the most frequent complication after hip surgery, followed by low limb embolism and malnutrition. The mortality is dramatically greater in patients over 80 years old than those below, and major causes are acute renal failure, multiple organ dysfunction syndrome and mental deterioration. Multidisciplinary consultations and mental assessment are encouraged in patients over 80 years old after hip trauma and surgery. Hip fractures in octogenarians and nonagenarians deserve special attention because of their advanced age and comorbidities.</p>

Association study on single nucleotide polymorphisms in HTRA1 gene and rheumatoid arthritis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the association between the single nucleotide polymorphisms (SNPs) in the high-temperature requirement A-1 (HTRA1) gene and rheumatoid arthritis (RA) in Chinese Han population.</p><p><b>METHODS</b>Five SNPs in the HTRA1 gene (rs2014307, rs2248799, rs2300433, rs714816 and rs2268356) were genotyped by ABI Snapshot method in Han Chinese cohort composed of 344 patients with RA and 288 healthy controls. The serum rheumatoid factor (RF) and C-reactive protein (CRP) of the patients were determined by endpoint nephelometry method.</p><p><b>RESULTS</b>Genotypes of all the five SNPs in the HTRA1 gene were not significantly different between the RA patients and controls (P> 0.05). Haplotypes generated by these five SNPs did not show significantly difference between the two groups either (P> 0.05). Serum RF levels in the RA patients had no significant difference among the genotypes for four SNPs (rs2014307, rs2248799, rs714816, and rs2268356) in the HTRA1 gene, while RF levels in the RA patients with genotypes AA+AG of the rs2300433 locus were significantly higher than that in genotype GG carriers (P< 0.05). Serum CRP levels in the RA patients had no significant difference among the genotypes for all the five SNPs.</p><p><b>CONCLUSION</b>Author's results suggested that although the five SNPs in the HTRA1 gene were not associated with RA in Chinese Han population, RF levels in the RA patients with genotypes AA and AG in the rs2300433 locus were significantly higher than the GG carriers. The HTRA1 role in RF regulation needs to be further investigated.</p>

Relationship between surgical time and postoperative complications in senile patients with hip fractures / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To elucidate the relationship between surgical time and postoperative complications in senile patients with hip fractures, and try to find out other factors which are related to these complications.</p><p><b>METHODS</b>Sixty-two patients, 28 males aged from 65 to 72 years with a mean age of 76.3 years and 34 females aged from 65 to 95 years with a mean age of 78.1 years, who had undergone orthopedic surgery because of hip fractures, were enrolled in a retrospective cohort study. The surgical time and pattern, the type of fracture, preoperative comorbidities, American Society of Anesthesiologists (ASA) score and the volume of blood transfusion during operation were obtained from these patients who were followed up by telephone calls for postoperative complications. All the patients were followed up at least for 1 year and were divided into subgroups according to their clinical characteristics and the results were analyzed by the Statistical Analysis System software.</p><p><b>RESULTS</b>There was no significant difference in the morbidity of postoperative complications with the gender, age, surgical time and pattern, or ASA score. There was significant difference in the morbidity of postoperative complications related to preoperative comorbidities and the volume of blood transfusion. There was a significant causality between preoperative comorbidities and postoperative complications. The morbidity of postoperative complications was 1.651 times higher in patients with preoperative comorbidities than those without.</p><p><b>CONCLUSIONS</b>There is no relationship between the surgical time and postoperative complications in senile patients who received surgery for hip fracture within 1 year. No correlation is found between the postoperative complications and gender, age, type of fracture, surgical pattern, ASA score and the volume of blood transfusion. Preoperative comorbidities are an independent predictor for postoperative complications.</p>

Disease gene screening of known loci in a Chinese family with autosomal dominant retinitis pigmentosa / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To map the disease-causing gene in a Chinese family with autosomal dominant retinitis pigmentosa.</p><p><b>METHODS</b>Twenty-seven micro-satellite markers were randomly selected from the region around the known loci of causative genes, and haplotypes were determined by ABI3100 genetic analyzer. Two-point linkage analysis was performed using MLINK.</p><p><b>RESULTS</b>The Lod score of each marker vs adRP was below 1.</p><p><b>CONCLUSION</b>The phenotype of this family may not be caused by mutation of the known disease-causing genes.</p>

Linkage analysis of a family with familial hypertriglyceridemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To perform linkage analysis and mutation screening in a Chinese family with familial hpertriglyceridemia (FHTG).</p><p><b>METHODS</b>Thirty-two family members including 12 hypertriglyceridemia patients participated in the study. Genotyping and haplotype analysis for 22 subjects were performed using short tandem repeat (STR) microsatellite polymorphism markers on 16 candidate genes and/or loci related to lipid metabolism. Two of the sixteen known candidate genes, APOA2 and USF1 were screened for mutation by direct DNA sequencing.</p><p><b>RESULTS</b>No linkage was found between the candidate genes/loci of APOA5, LIPI, RP1, APOC2, ABC1, LMF1, APOA1-APOC3-APOA4, LPL, APOB, CETP, LCAT, LDLR, APOE and the phenotype in this family. The two-point Lod scores (theta =0) were all less than-1.0 for all the markers tested. Linkage analysis suggested linkage to chromosome 1q23.3-24.2 between the disease phenotype and STR marker D1S194 with a two-point maximum Lod score of 2.44 at theta =0. Fine mapping indicated that the disease gene was localized to a 5.87 cM interval between D1S104 and D1S196. No disease-causing mutation was detected in the APOA2 and USF1 genes.</p><p><b>CONCLUSION</b>The above mentioned candidate genes were excluded as the disease causing genes for this family. The results implied that there might be a novel gene/locus for FHTG on chromosome 1q23.3-1q24.2.</p>

The trend of cancer mortality from 1988 to 2005 in Kaifeng county, China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the time trends of cancer mortality among residents in Kaifeng county, Henan province.</p><p><b>METHODS</b>Data on cancer mortality from the vital registration system in Kaifeng county from 1988 to 2005 was analyzed. A total of 9543 death records (5974 males and 3567 females) due to malignant tumors were studied. A two-year-period age-specified standardized mortality rates were directly adjusted by the world standard population, and the annual percentage change (APC) of mortality were estimated by a linear logarithm regression.</p><p><b>RESULTS</b>The crude cancer death rate for male was 95.09/100,000 and its age-standardized death rate was 117.41/100,000. While, the crude cancer death rate for female was 59.13/100,000 and the age-standardized death rate was 57.15/100,000. There was a significant growth tread for lung cancer (APC: 6.54%), liver cancer (5.07%) in males and breast cancer (7.04%) in females in the groups aged over 18. On the contrary, the decreasing treads for esophageal cancer in both of sexes (-7.09%, -13.53%) were also observed in this study. Meanwhile, there was no other significant changes in the trend, either in the tumor sites or mortality, was observed.</p><p><b>CONCLUSION</b>In the past two decades, there has been a significant increasing trend for cancer mortality in Kaifeng county, of Henan Province. Hence, it is necessary to enhance epidemiological survey to identify risk factors at the earlier stages.</p>