Analysis of KIF1A gene variant in a Chinese pedigree affected with Spastic paraplegia type 30 / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese pedigree affected with hereditary spastic paraplegia type 30 (HSP30).@*METHODS@#A proband presented at the Second Hospital of Shanxi Medical University in August 2021 was selected as the study subject. The proband was subjected to whole exome sequencing, and candidate variant was verified by Sanger sequencing and bioinformatic analysis.@*RESULTS@#The proband was found to have harbored a heterozygous c.110T>C variant in exon 3 of the KIF1A gene, which can cause substitution of isoleucine by threonine at position 37 (p.I37T) and alter the function of its protein product. The same variant was not found in his parents, elder brother and elder sister, suggesting that it has a de novo origin. Based on the guidelines of the American College of Medical Genetics and Genomics (ACMG), the variant was rated as likely pathogenic (PM2_Supporting+PP3+PS2).@*CONCLUSION@#The c.110T>C variant of the KIF1A gene probably underlay the HSP30 in the proband. Above finding has enable genetic counseling for this family.

Analysis of PKD2 gene variant and protein localization in a pedigree affected with polycystic kidney disease / 中华医学遗传学杂志

OBJECTIVE@#To detect the mutation site in a pedigree affected with autosomal dominant polycystic kidney disease (ADPKD) and verify its impact on the protein function.@*METHODS@#Peripheral blood samples were collected from the proband and his pedigree members for the extraction of genomic DNA. Mutational analysis was performed on the proband through whole-exome sequencing. Suspected variant was verified by Sanger sequencing. A series of molecular methods including PCR amplification, restriction enzyme digestion, ligation and transformation were also used to construct wild-type and mutant eukaryotic expression vectors of the PKD2 gene, which were transfected into HEK293T and HeLa cells for the observation of protein expression and cell localization.@*RESULTS@#The proband was found to harbor a c.2051dupA (p. Tyr684Ter) frame shift mutation of the PKD2 gene, which caused repeat of the 2051st nucleotide of its cDNA sequence and a truncated protein. Immunofluorescence experiment showed that the localization of the mutant protein within the cell was altered compared with the wild-type, which may be due to deletion of the C-terminus of the PKD2 gene.@*CONCLUSION@#The c.2051dupA (p. Tyr684Ter) mutation of the PKD2 gene probably underlay the pathogenesis of ADPKD in this pedigree.

Identification of a novel SOD1 variant in a Chinese patient with amyotrophic lateral sclerosis / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese patient with amyotrophic lateral sclerosis (ALS).@*METHODS@#Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA. Genetic variant was identified by whole exome sequencing. Candidate variant was verified by Sanger sequencing of his parents and healthy controls.@*RESULTS@#The patient was found to harbor a heterozygous c.420C>G (p.Asn140Lys) variant of the SOD1 gene. The same variant was not detected in his parents and 100 healthy controls. The variant has not been included in HGMD, dbSNP and other databases.@*CONCLUSION@#The c.420C>G variant of the SOD1 gene may underlie the ALS in this patient. Above finding has enriched the spectrum of SOD1 gene variants.

Instability of Mitochondrial DNA D-loop Region Genes in Patients with Leukemia / 中国实验血液学杂志

OBJECTIVE@#To study the instability of mitochondrial DNA（mt DNA） D-loop region genes in patients with Leukemia.@*METHODS@#The HV-1 and HV-2 regions of D-loop region in 24 patients with leukemia were amplificated and sequenced, then their results were compared with revised Cambridge reference sequence (rCRS) and Databank mtDB. The mutation rate was detected by SPSS 22.0 statistics software.@*RESULTS@#The total mutation rate in patients was 95.83% (23/24), the detection showed 82 mutated genes, out of which 47 (57.32%) mutated genes located in HV-1 region, 35 (42.68%) mutated genes in HV-2 region. The comparison showed that the mutation rate in untreated (UT) group and treated (T) group of AML patients was (2.37±0.82)×10 and (4.76±2.45)×10 respectively(P＜0.01), the mutation rate in PR and CR groups of treated AML patients was (5.10±2.56)×10 and (4.51±2.51)×10 respectively (P＜0.05), the comparison among M3 group showed that the mutation rates in UT, PR and CR groups were (2.55±0.63)×10, (5.37±3.41)×10 and (3.71±1.65)×10 respectively (P＞0.05).@*CONCLUSION@#The more high mutation rate and many kinds of mutation types exist in D-loop region, suggesting that the genes in D-loop region display the more strong instability, the chemotherapy may aggravate the instability of genes in D-loop region.

Screening for EXT1 and EXT2 gene mutations in a ethnic Han Chinese family from Shanxi with hereditary multiple exostoses / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen for potential mutations in an ethnic Han Chinese family from Shanxi with hereditary multiple exostoses.</p><p><b>METHODS</b>Polymerase chain reaction and DNA sequencing were used to screen potential mutations in EXT1 and EXT2 genes.</p><p><b>RESULTS</b>For EXT1 gene, two synonymous mutations (P477P and E587E), three intronic mutations (c.1537 -48A>G, c.1721 +203A>G and c.1722 -103C>G) were detected. For EXT2 gene, five intronic mutations (c.-29 -148A>T, c.1080 -18T>A, c.1336 -93C>T, c.1526 -166C>T, and c.1526 -195C>T) were identified. Among these, EXT1 P477P, EXT1 E587E and EXT2 c.1080 -18T>A are polymorphisms listed by Multiple Osteochondroma Mutation Database, whilst the other 7 sites have not been reported.</p><p><b>CONCLUSION</b>No mutations have been found among all exons of the EXT1 and EXT2 genes in this family. Linkage analysis is necessary for identifying the cause of this disease.</p>

An experimental study on chemoprevention of esophageal adenocarcinoma by celecoxib, a selective cyclooxygenase-2 inhibitor / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the feasibility of chemoprevention of esophageal adenocarcinoma by celecoxib, a selective cyclooxygenase-2(COX-2) inhibitor using a rat model.</p><p><b>METHODS</b>Rats were divided into 3 groups: model group, celecoxib group, and control group. The rat surgical model was established by performing a gastrojejunostomy plus an esophagojejunostomy 5 mm distal to the gastrojejunal anastomosis. Twenty-eight weeks after surgery, all the animals were sacrificed and the pathological changes in the esophagus were examined macroscopically. COX-2 expression was analyzed by immunohistochemistry. Prostaglandin E2(PGE2) level was measured by enzyme-linked immunosorbent assay(ELISA).</p><p><b>RESULTS</b>The incidence of Barrett's esophagus and esophageal adenocarcinoma in the model group was 84% and 57% respectively, significantly higher than those in the control group(P<0.01). The incidence of esophageal adenocarcinoma in the celecoxib-treated group was significantly lower than that in the model group(P<0.01), and no esophageal adenocarcinoma was detected in the control group. COX-2 expression was detected in 100% of reflux esophagitis, Barrett esophagus and esophageal adenocarcinoma, but not found in the normal tissue from the esophagus and the jejunum(P<0.01). The PGE2 level in the esophageal tissue in the model group was significantly higher than that in the control group(P<0.01). Rats in the celecoxib-treated group had significantly lower PGE2 level than that in the model group(P<0.01). The PGE2 levels were significantly higher in rats with cancer than those without cancer(P<0.01).</p><p><b>CONCLUSION</b>Celecoxib successfully prevents the development of esophageal adenocarcinoma in a rat surgical model with mixed reflux of acid and duodenal juice and significantly decreases the risk of Barrett esophagus developing esophageal adenocarcinoma. COX-2 maybe an effective selective target of chemoprevention for esophageal adenocarcinoma.</p>

Detection of exon 7 mutations of PAH gene in classical phenylketonuria by high-resolution melting analysis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a simple, rapid, inexpensive and sensitive method for detecting hot region for mutations in exon 7 of PAH gene.</p><p><b>METHODS</b>High-resolution melting (HRM) technology was used to detect a c.728G>A mutation in exon 7 in 88 patients with classical type phenylketonuria. Suspected mutations were validated by direct DNA sequencing.</p><p><b>RESULTS</b>The results detected by HRM are in good agreement with the results obtained by direct sequencing.</p><p><b>CONCLUSION</b>HRM analysis is a simple, rapid, inexpensive and sensitive method for detecting hot mutational region in exon 7 of PAH gene.</p>

Risk factors of surgical interventions in patients with esophageal foreign bodies / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the risk factors of surgical interventions in patients with esophageal foreign bodies(EFBs).</p><p><b>METHODS</b>Forty-three cases with EFBs underwent surgical interventions from January 1962 to January 2011 in Department of Thoracic Surgery at the Tangdu Hospital were enrolled in the study group. Forty three cases with EFBs who received non-surgical treatment in the same hospital and during the same period matched by gender and age(the age difference between the paired patients was 5 years or less) were selected as the control group. Clinical data including type of EFBs, duration of impaction, location of EFBs, complications associated with EFBs were collected. Univariate and multivariate conditional logistic analysis were used for risk factors analysis.</p><p><b>RESULTS</b>Univariate conditional logistic analysis showed that type of EFBs, duration of impaction, location of EFBs, and complications associated with EFBs were risk factors of surgical interventions in patients with EFBs(P<0.05). Further multivariate conditional logistic analysis showed that type of EFBs(sharp EFBs) was the independent factor of surgical interventions in patients with EFBs(P=0.043; OR=29.228, 95% CI:1.114-766.686).</p><p><b>CONCLUSIONS</b>Physician should pay more attention to patients with sharp EFBs. Whether the patients need surgical interventions or not should base on factors including duration of impaction, location of EFBs, and complications associated with EFBs.</p>

Study on the mutations of phenylalanine hydroxylase gene in patients with phenylketonuria in Shanxi province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the mutations in exons 3, 6, 7, 11 and 12 of the phenylalanine hydroxylase gene (PAH) in Shanxi population.</p><p><b>METHODS</b>The mutations in exons 3, 6, 7, 11 and 12 and flanking sequences of PAH gene were detected by PCR-DNA sequencing, in 59 patients with phynelketonuria(PKU) and 100 healthy children from Shanxi province.</p><p><b>RESULTS</b>By sequence analysis, three single nucleotide polymorphism (SNP) Q232Q (CAA>CAG), V245V (GTG>GTA) and L385L (CTG>CTC) were detected in both the patients and healthy children, with the frequencies of nt 696, 735 and 1155 of the PAH cDNA up to 96.2%, 76.1% and 7.6% in patients respectively, and 97.0%, 77.3% and 8.3% respectively in the healthy controls. In addition, 72 different mutations accounting for 61.0% of mutant alleles were identified in the patients only. In exon 3, R111X, H64>TfsX9 and S70 del were found accounting for 5.1%, 0.8% and 0.8%; EX6-96A>G in exon 6 was found accounting for 10.2%. In exon 7, R243Q was the highest incidence accounting for 12.7%, followed by Ivs7+2 T>A(5.1%) and T278I(2.5%); the lowest incidences were G247V, R252Q, L255S, R261Q and E280K accounting for 0.8 %, respectively. In exon 11, Y356X (5.9%) and V399V (5.1%) were found; in exon 12, R413P and A434D were found accounting for 5.9% and 2.5%. In total, 9 missense mutations, 3 splice site mutations, 2 nonsense mutations and 2 deletions were included in 16 kinds of different mutations.</p><p><b>CONCLUSION</b>The mutation characteristics and distribution in exons 3, 6, 7, 11 and 12 of the PAH gene have been identified, and it suggested that the EX6-96A>G and R243Q were the hot spots of PAH gene mutations in Shanxi PKU population.</p>

Screening metastasis-associated genes from anoikis resistant A549 lung cancer cells by human genome array / 中国肺癌杂志

<p><b>BACKGROUND AND OBJECTIVE</b>As a barrier to metastases, cells normally undergo apoptosis after they lose contact with their extra cellular matrix (ECM). This process has been termed "anoikis". Tumour cells that acquire malignant potential have developed mechanisms to resist anoikis and thereby survive after detachment from their primary site while traveling through the lymphatic and circulatory systems. This "anoikis resistance" is considered the first step to tumor metastases. The aim of this study was to screen metastasis-associated genes from anoikis resistant and adherent growth A549 lung cancer cell by Human Genome Array.</p><p><b>METHODS</b>Establish anoikis resistant A549 lung cancer cell lines by using poly-hydroxyethyl methacrylate resin processed petri dishes, which causes cell free from adherent. The different expressed gene between anoikis resistant A549 cell and adherent growth A549 cell was tested using human V2.0 whole-genome oligonucleotide microarray, a product of Capitalbio Corporation, Beijing. Screen metastasis-associated genes.</p><p><b>RESULTS</b>745 different expressed genes were screened, including 63 highly metastasis-associated genes.</p><p><b>CONCLUSION</b>The successfully established anoikis resistant A549 cell lines and screened different expressed genes provide us basis for further research on metastasis of lung cancer.</p>

Expression and significance of P-gp/mdr1 mRNA, MRP and LRP in non-Hodgkin's lymphoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the expression and clinical significance of P-glycoprotein (P-gp)/mdr1mRNA, multidrug resistance-associated protein (MRP) and lung resistance protein (LRP) in newly diagnosed non-Hodgkin's lymphoma.</p><p><b>METHODS</b>mdr1 mRNA of in 41 patients with non-Hodgkin's lymphoma was assayed by semi-quantitative RT-PCR. The expressions of P-gp, MRP and LRP proteins in lymph node viable blasts were identified by flow cytometry. The results were compared with those obtained from control cases, and the correlation of the changes with clinical outcomes was analyzed.</p><p><b>RESULTS</b>(1) Among the 41 cases, the positive expression of P-gp protein was detected in 8 cases, MRP in 7 cases, LRP in 15 cases, and mdr 1 mRNA in 11 cases. (2) The P-gp and LRP levels in NHL were significantly higher than those in control group, but MRP wasn't. The P-gp over-expression was significantly associated with mdr1mRNA (r = 0.396, P = 0.01). No correlation was showed among the expressions of P-gp, MRP and LRP. (3) Patients with P-gp expression had a poorer outcome of chemotherapy than those with P-gp-negative (P = 0.005). P-gp expression was significantly associated with higher clinical stage (P = 0.046) and elevated serum lactate dehydrogenase level (P = 0.032), but not associated with malignant degree (P = 0.298). MRP had no impact on the outcome of chemotherapy (P = 0.212), and wasn't significantly associated with higher clinical stage (P = 0.369), elevated LDH (P = 0.762) and higher malignant degree (P = 0.451). Patients with LRP expression had a poorer outcome of chemotherapy than those LRP-negative (P = 0.012). LRP expression was significantly associated with higher clinical stage (P = 0.0019), elevated LDH (P = 0.02) and higher malignant degree (P = 0.01).</p><p><b>CONCLUSION</b>The data of this study indicate that P-gp and LRP expressions but not MRP expression are important in the mechanism of drug resistance associated with a poor clinical outcome in previously untreated NHL.</p>

Mechanism of C-reactive protein on proliferation of multiple myeloma U266 cells / 中国实验血液学杂志

This study was purposed to investigate the mechanism of C-reactive protein (CRP) on proliferation of U266 cells. The human multiple myeloma cell line U266 was incubated with human CRP (0, 5, 10, 20 mg/L) for 24 hours, then the proliferation level of U266 cells was detected by using blood analyser. The mRNA expressions of survivin and HSP90alpha were examined by RT-PCR. The results showed that the proliferation ratio was increased, as compared with the control group (p<0.05); furthermore, the mRNA levels of survivin and HSP90alpha were up-regulated in proportion to the increased CRP concentrations. There was significant correlation between expression of survivin and HSP90alpha (r=0.737, p<0.0001) in incubated cells. It is concluded that CRP can stimulate the proliferation of MM cells directly by up-regulating the expression of survivin and HSP90alpha in MM cells. CRP can be regarded as a potential target for MM treatment.

Association of MMP1 -1607(1G>2G)single nucleotide polymorphism with susceptibility to lung cancer in Northwestern Chinese population of Han nationality / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Matrix metalloproteinase 1 (MMP1) plays an important role in the development of lung cancer. This study was to investigate the relation to associate the single nucleotide polymorphism(SNP)in MMP1 gene with the susceptibility to lung cancer in Northwestern Chinese population of Han nationality.</p><p><b>METHODS</b>By using the methods of polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP), MMP1 -1607(1G>2G) polymorphisms in 150 patients with lung cancer, and 200 healthy controls were detected to evaluate the relationship between different genotypes and susceptibility of lung cancer.</p><p><b>RESULTS</b>Individuals with 2G/2G genotype had 1.77 fold risk suffering from lung cancer, when compared with ones with 1G/2G and 1G/1G genotypes. Smokers with 2G/2G genotype exhibited 3.20 fold elevated risk for lung cancer (OR 3.20; 95% CI 1.50-6.82).</p><p><b>CONCLUSION</b>The -1607(1G>2G) in promoter region of MMP1 is associated with susceptibility to lung cancer in Northwestern Chinese population of Han nationality. The genotype 2G/2G enhances the susceptibility to lung cancer.</p>

Effect of ischemic preconditioning on cytokines during lung ischemia-reperfusion injury / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the protective role of ischemic preconditioning (IPC) during lung ischemia-reperfusion (I/R) injury and its influence on inflammatory cytokine production.</p><p><b>METHODS</b>In vivo I/R injury of rabbit was induced by blocking hilum of the left lung. The wet/dry ratio of the lung, lung permeability index and neutrophils percentage in bronchoalveolar lavage fluid (BALF) were detected as indexes of the lung injury. Serum levels of tumor necrosis factor alpha (TNFalpha), interleukin-6 (IL-6) and interleukin-8 (IL-8) were also detected using enzyme-linked immunosorbent assay. The protective role of IPC and its influence on inflammatory cytokine production were observed.</p><p><b>RESULTS</b>The wet/dry ratio of the lung, lung permeability index and neutrophils percentage in BALF of I/R group were 9.73 +/- 1.14, (41.62 +/- 5.77) x 10(-4) and (58.1 +/- 10.0)% respectively. The IPC group indexes were 6.23 +/- 0.69, (20.31 +/- 4.03) x 10(-4) and (23.8 +/- 5.2)% respectively. There was a significant difference between the two groups (P < 0.01). Serum levels of TNFalpha, IL-6 and IL-8 of I/R group were (0.9078 +/- 0.1062), (0.2137 +/- 0.0598) and (0.7211 +/- 0.0979) ng/ml respectively. The IPC group indexes were (0.7478 +/- 0.0843), (0.1271 +/- 0.0089) and (0.5903 +/- 0.0746) ng/ml respectively, significantly lower than that of I/R group (P < 0.01).</p><p><b>CONCLUSIONS</b>Lung IPC has a marked protection effect against I/R injury. The effect was related to its inhibition of inflammatory cytokines such as TNFalpha, IL-6 and IL-8, thus reducing activation and infiltration of neutrophils.</p>

Effect of anti-VEGF₁₆₅ ribozyme on biological behaviors of human lung adenocarcinoma cell / 中国肺癌杂志

<p><b>BACKGROUND</b>To investigate the biological effects of anti VEGF₁₆₅ ribozyme on human lung adenocarcinoma cell.</p><p><b>METHODS</b>Hammerhead ribozyme (VRz) against VEGF₁₆₅ gene transcripts (site 212) and its paired mutant ribozyme (mVRz) were designed and synthesized, and the cleavage activity of the ribozymes on target RNA in a cell-free system was observed. The replication-incompetent adenovirus-mediated eukaryotic expression vectors (rpAdVRz) containing VRz and mVRz gene were constructed and identified. Then the human lung adenocarcinoma cells (A549) were infected with recombinant adenovirus. The biological characteristics of A549 cell before and after infection in vitro were inspected by Northern blot, laser confocal imaging system analysis, flow cytometry and transmission electron microscopy.</p><p><b>RESULTS</b>VRz specifically and efficiently cleaved the VEGF₁₆₅ mRNA. The rpAdVRz was successfully constructed and infected A549 cell. The level of VEGF₁₆₅ expression decreased 87% in rpAdVRz infected cells compared with the other groups, but their biological characteristics were not influenced by the expression of the exogenous gene.</p><p><b>CONCLUSIONS</b>The adenovirus mediated hammerhead ribozyme against VEGF₁₆₅ can significantly decrease the expression of VEGF₁₆₅. This provides an experimental basis for human lung cancer gene therapy with antiangiogenesis method.</p>

Effect of transfection of p27(kipl) on lung cancer cells line A549 / 中国肺癌杂志

<p><b>BACKGROUND</b>To observe the suppressive effects of exogenous p27 gene on human lung cancer cell line A549.</p><p><b>METHODS</b>An adenovirus expression vector (pAd CMV-p27) containing 570 bp human full-length p27 cDNA was transfected into human lung cancer cell line A549. Expression of exogenous p27 gene was detected by dot-blot hybridization and laser co-focal system. MTT was adopted to measure the effects of exogenous p27 gene on cell cycle progression and cell features of the infected A549.</p><p><b>RESULTS</b>The mRNA and protein expression level of p27 was remarkably increased after transfecting with exogenous p27 gene. The apoptosis of infected A549 occurred and the progression of cell cycle was arrested in G1 phase.</p><p><b>CONCLUSIONS</b>p27(kipl) gene transfer may play a therapeutic role in the treatment of lung cancer.</p>