Genetic relatedness of selected clinical Vibrio cholerae O139 isolates from the southern coastal area of China over a 20-year period.

Vibrio cholerae O139 emerged as a causative agent of epidemic cholera in 1992 in India and Bangladesh, and was subsequently reported in China in 1993. The genetic relatedness and molecular characteristics of V. cholerae O139 in Guangdong Province, located in the southern coastal area of China, remains undetermined. In this study, we investigated 136 clinical V. cholerae O139 isolates from 1993 to 2013 in Guangdong. By conventional PCR, 123 (90·4%) isolates were positive for ctxB, ace and zot. Sequencing of the positive amplicons indicated 113 (91·7%) isolates possessed the El Tor allele of ctxB (genotype 3); seven carried the classical ctxB type (genotype 1) and three harboured a novel ctxB type (genotype 5). With respect to tcpA, 123 (90·4%) isolates were positive for the El Tor allele. In addition, pulsed-field gel electrophoresis (with NotI digestion) differentiated the isolates into clusters A and B. Cluster A contained seven of the non-toxigenic isolates from 1998 to 2000; another six non-toxigenic isolates (from 1998 and 2007) and all of the toxigenic isolates formed cluster B. Our results suggest that over a 20-year period, the predominant O139 clinical isolates have maintained a relatively tight clonal structure, although some genetic variance and shift has occurred. Our data highlight the persistence of toxigenic V. cholerae O139 in clinical settings in the southern coastal area of China.

[Research progress in mesenchymal stem cell-related therapies in the treatment of temporomandibular joint osteoarthritis].

Temporomandibular joint osteoarthritis (TMJOA) is a kind of organic disease with synovial inflammation, cartilage degeneration and subchondral bone remodeling as the main pathological changes. The current treatment is mainly to relieve symptoms, but cannot completely stop the progression of the disease. Mesenchymal stem cells (MSC) have multi-lineage differentiation potential and have good prospects in the repair therapy of TMJOA. Intra-articular injection of MSC from bone marrow, adipose, umbilical cord, dental pulp, etc. has been shown to be effective in numerous animal studies. The above exogenous MSCs can also be used as seed cells to participate in tissue engineering and repair more severe defects. Recent studies have shown that exosomes are important mediators of MSC action and have some potential in the treatment of TMJOA. As the mechanisms of TMJOA are further investigated, there is some prospect that endogenous repair capacity can be activated by local injection of relevant drugs targeting the resident stem cells in the joint.

[Etiological characterization of invasive non-typhoid Salmonella strains in Guangdong Province from 2018 to 2022].

Objective: To understand the serotype distribution, drug resistance and molecular characterization of invasive non-typhoid Salmonella (iNTS) in Guangdong Province from 2018 to 2022 and provide scientific evidence for the prevention and treatment of blood flow infection caused by Salmonella. Methods: Serological identification, antimicrobial susceptibility testing, multilocus sequence typing (MLST), and whole genome sequencing were performed on Salmonella isolated from blood and stool samples in Guangdong from 2018 to 2022. Simultaneously, annotated the sequencing results for drug resistance genes and virulence factors by a microbial gene annotation system. Results: The 136 iNTS strains were divided into 25 serotypes, and Salmonella enteritidis accounted for 38.24% (52/136). The OR of other iNTS serotypes were calculated with Salmonella typhimurium as the control. The OR values of Oreninburg, Rysson, and Pomona serotypes were the highest, which were 423.50, 352.92, and 211.75, respectively. The drug resistance rate of iNTS was 0.74%-66.91%, which was lower than that of non-iNTS (3.90%-77.21%). The main iNTS of drug resistance were ampicillin and tetracycline, with resistance rates of 66.91% (91/136) and 50.00% (68/136), respectively, while the resistance rates to ciprofloxacin (5.88%,8/136), ceftazidime (5.88%,8/136), gentamicin (5.13%,7/136) and cefoxitin (0.74%, 1/136) were relatively low. iNTS carried a variety of drug-resistance genes and virulence factors, but no standard virulence factor distribution has been found. MLST cluster analysis showed that iNTS was divided into 26 sequence types, and ST11 accounted for 38.24% (52/136). Conclusions: The iNTS strains in Guangdong were dominated by Salmonella enteritidis, of which three serotypes, Oreninburg, Rison, and Pomona, may be associated with a higher risk of invasive infection during 2018 to 2022. iNTS was sensitive to clinical first-line therapeutic drugs (cephalosporins and fluoroquinolones), with highly diverse sequences and clear phylogenetic branches. ST11 was the local dominant clone group.

[Consensus of experts on standard items of the cohort construction and quality control of temporomandibular joint diseases (2024)].

Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients'suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aimsto establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. Temporomandibular joint disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.

[Research progress in the pathologic mechanism of temporomandibular joint osteoarthritis].

Temporomandibular joint osteoarthritis (TMJOA) is a kind of degenerative lesion in temporomandibular joint disorders. The pathogenesis of TMJOA is complex. Among them, excessive mechanical stress plays an important role, which can cause a series of pathological changes, including synovial inflammation, chondrocyte death, osteogenic and adipogenic differentiation, degradation of extracellular matrix of cartilage, angiogenesis, osteochondral interface sclerosis, subchondral bone remodeling, degeneration of articular discs, etc. In the pathogenesis of TMJOA, it involves multiple signal pathways and epigenetic regulation, which may become potential therapeutic targets of TMJOA. In this paper, the research progress on the pathologic mechanism of TMJOA in recent years is reviewed to provide a basis for finding new therapeutic targets of TMJOA.

Study on the lateral pterygoid muscle status after artificial temporomandibular joint replacement.

The purpose of this study was to explore the status of the lateral pterygoid muscle (LPM) after detachment in artificial temporomandibular joint replacement (TJR) surgery. Patient clinical and computed tomography imaging data were collected before and after unilateral artificial TJR with LPM detachment. The volume of the LPM on the operated and unoperated sides was measured before and after surgery (at 1, 3, 6, 12 months) using ProPlan CMF 3.0 software. The volumes of the LPM on both sides, the patient's mandibular movements, quality of life (QoL), and pain and diet scores (visual analogue scales) were evaluated and compared at the different follow-up stages. Ten patients were included in the study. After surgery, the volume of the operated LPM was significantly reduced to 60.78% at 3 months (P=0.007), and gradually stabilized to 51.58% at 6 months (P=0.025) and 54.68% at 1 year postoperative (P=0.002). There were no significant LPM volume changes on the unoperated side (P=0.67). Lateral movement of the operated joint was significantly reduced (P=0.021) and correlated with the LPM volume change after surgical detachment (P=0.042). The LPM shrank after detachment in the artificial TJR surgery and the muscle detachment affected the movement of the replaced joint.

[Clinical analysis of seven cases of H1N1 influenza-associated encephalopathy in children].

Objective: To investigate the clinical manifestations, diagnosis, and treatment of H1N1 influenza A-associated encephalopathy (IAE) in children. Methods: The clinical manifestations, laboratory tests, cranial magnetic resonance imaging (MRI), electroencephalography (EEG) examinations and treatments of seven children with H1N1 IAE hospitalized in Guangzhou Women and Children's Medical Center from December 2018 to January 2019 were retrospectively analyzed. Results: Five of the seven children with H1N1 IAE were female. The age at admission was 4 years and 5 months (range 7 months-9 years). Neurological symptoms occurred simultaneously or early (0-3 days) after the flu-like symptom appeared. The main clinical manifestations of neurological symptoms were seizures (repeated seizures in five cases and status convulsion in two cases, including one case of unexpected fever and repeated seizures in a nine-year old girl) accompanied with altered consciousness (drowsiness in five cases and coma in two cases). Cranial MRI in three cases displayed multifocal lesions, mainly in the bilateral thalamus, brainstem and cerebellar hemisphere. MRI also showed reversible splenial lesion in the corpus callusumin in three cases. EEG tracings were characterized by diffuse slow wave activity in four cases, and status epilepticus was monitored in one case. All the 7 cases were treated with oral oseltamivir. Three cases were treated with pulsed methylprednisolone and intravenous immunoglobulin. One case was treated with intravenous immunoglobulin alone and all the patients received oral oseltamivir. All the patients survived, with three patients had minor neurological sequelae at discharge. Conclusions: The main clinical manifestations of H1N1 IAE are seizures and altered consciousness. Cranial MRI combined with EEG is helpful for early diagnosis. Intravenous immunoglobulin and (or) methylprednisolone should be considered for severe cases.

[Circulation and etiological characterization of Salmonella enterica serotype in human in Guangdong province, 2007-2016].

Objective: To understand the circulation, drug resistance and molecular characteristics of Salmonella1, 4, [5], 12: i:- in human in Guangdong province. Methods:Salmonella1, 4, [5], 12: i:- isolated from diarrhea patients in Guangdong during 2007-2016 were detected for drug resistance, genes and PFGE characteristics. Results: A total of 2 960 strains Salmonella1, 4, [5], 12: i: - were isolated from human diarrhea cases during this period. The positive rates of the isolation increased year by year. The male to female ratio of the infection cases was 1.58∶1, and the infection mainly occurred in infants and young children. Except imipenem, Salmonella1, 4, [5], 12: i: - was resistant to other 17 antibiotics to some extent. The drug resistant rates to ceftazidime, cefotaxime and ciprofloxacin increased from 2011 to 2016. Multi-drug resistance was serious, for example, the multi-drug resistant strains with ASSuT accounted for 70.62% (435/616) and the multi-drug resistant strains with ACSuGSTTm accounted for 27.11% (167/616). The lack of fljA, fljB and hin genes, as well as the retaining of iroB, STM2740, STM2757 genes, resulted in the unable expression of FljBenx gene with 8 different defection profiles. There were 934 different PFGE patterns observed in 2 347 strains, which displayed a relatively large fingerprint polymorphism. The major PFGE pattern was JPXX01. GD0226, which was found in 97 strains, accounting for 4.13% (97/2 347). The PFGE patterns in 168 Salmonella1, 4, [5], 12: i: - strains were consistent with that of Salmonella typhimurium. Conclusions:Salmonella1,4,[5], 12: i: - strains has become the major serotype of Salmonella that cause diarrhea in human in Guangdong. The multi-drug resistance of Salmonella1,4, [5], 12: i: - was serious, and since the defection of fljA, fljB and hin genes, the expression of FljBenx protein failed. The PFGE results were diverse, which displayed polymorphism in inheritance.

Evidence for a G2 checkpoint in p53-independent apoptosis induction by X-irradiation.

The p53 tumor suppressor gene is thought to be required for the induction of programmed cell death (apoptosis) initiated by DNA damage. We show here, however, that the human promyelocytic leukemia cell line HL-60, which is known to be deficient in p53 because of large deletions in the p53 gene, can be induced to undergo apoptosis following X-irradiation. We demonstrate that the decision to undergo apoptosis in this cell line appears to be made at a G2 checkpoint. In addition, we characterize an HL-60 variant, HCW-2, which is radioresistant. HCW-2 cells display DNA damage induction and repair capabilities identical to those of the parental HL-60 cell line. Thus, the difference between the two cell lines appears to be that X-irradiation induces apoptosis in HL-60, but not in HCW-2, cells. Paradoxically, HCW-2 cells display high levels of expression of bax, which enhances apoptosis, and no longer express bcl-2, which blocks apoptosis. HCW-2 cells' resistance to apoptosis may be due to the acquisition of expression of bcl-XL, a bcl-2-related inhibitor of apoptosis. In summary, apoptosis can be induced in X-irradiated HL-60 cells by a p53-independent mechanism at a G2 checkpoint, despite the presence of endogenous bcl-2. The resistance shown by HCW-2 cells suggests that bcl-XL can block this process.

Ku86 defines the genetic defect and restores X-ray resistance and V(D)J recombination to complementation group 5 hamster cell mutants.

X-ray-sensitive hamster cells in complementation groups 4, 5, 6, and 7 are impaired for both double-strand break repair and V(D)J recombination. Here we show that in two mutant cell lines (XR-V15B and XR-V9B) from group 5, the genetic defects are in the gene encoding the 86-kDa subunit of the Ku autoantigen, a nuclear protein that binds to the double-stranded DNA ends. These mutants express Ku86 mRNA containing deletions of 138 and 252 bp, respectively, and the encoded proteins contain internal, in-frame deletions of 46 and 84 amino acids. Two X-ray-resistant revertants of XR-V15B expressed two Ku86 transcripts, one with and one without the deletion, suggesting that reversion occurred by activation of a silent wild-type allele. Transfection of full-length cDNA encoding hamster Ku86 into XR-V15B cells resulted in a complete rescue of DNA-end-binding (DEB) activity and Ku70 levels, suggesting that Ku86 stabilizes the Ku70 polypeptide. In addition, cells expressing wild-type levels of DEB activity were fully rescued for X-ray resistance and V(D)J recombination, whereas cells expressing lower levels of DEB activity were only partially rescued. Thus, Ku is an essential component of the pathway(s) utilized for the resolution of DNA double-strand breaks induced by either X rays or V(D)J recombination, and mutations in the Ku86 gene are responsible for the phenotype of group 5 cells.

[Development and preliminary clinical application of home-made temporomandibular joint prostheses].

Objective: To explore the clinical application of home-made temporomandibular joint (TMJ) prostheses. Methods: Self-developed TMJ prostheses were applied for end stage osteoarthritis, TMJ ankylosis, condylar tumors after biomechanical examination. The stability of the prostheses toward bone was evaluated during operation. Post-operative CT scans and clinical follow-ups were taken to check changes of mouth opening, position of the prostheses, bone resorption around the screws, and heterotopic bone formation around artificial condyle. Results: There were 6 patients accepted 7 prostheses smoothly without infection and other complications. Post-operative follow-ups were from 6 to 13 months with a mean of 7.6 months. Mouth opening was significantly improved from (16.2±14.4) mm before operation to (31.5±4.8) mm during follow-ups (P=0.01). All prostheses were stable without screw loose and heterotopic bone formation. Conclusions: Home-made TMJ prostheses were stable after primary clinical application, but the long-term results need to be observed.

Oncogenic activity of amplified miniature chromosome maintenance 8 in human malignancies.

Miniature chromosome maintenance (MCM) proteins play critical roles in DNA replication licensing, initiation and elongation. MCM8, one of the MCM proteins playing a critical role in DNA repairing and recombination, was found to have overexpression and increased DNA copy number in a variety of human malignancies. The gain of MCM8 is associated with aggressive clinical features of several human cancers. Increased expression of MCM8 in prostate cancer is associated with cancer recurrence. Forced expression of MCM8 in RWPE1 cells, the immortalized but non-transformed prostate epithelial cell line, exhibited fast cell growth and transformation, while knock down of MCM8 in PC3, DU145 and LNCaP cells induced cell growth arrest, and decreased tumour volumes and mortality of severe combined immunodeficiency mice xenografted with PC3 and DU145 cells. MCM8 bound cyclin D1 and activated Rb protein phosphorylation by cyclin-dependent kinase 4 in vitro and in vivo. The cyclin D1/MCM8 interaction is required for Rb phosphorylation and S-phase entry in cancer cells. As a result, our study showed that copy number increase and overexpression of MCM8 may play critical roles in human cancer development.

[Etiologic characteristics of Shigella sonnei strains isolated from some areas of Guangdong province and Guangxi Zhuang Autonomous Region of China, 2014-2016].

Objective: To investigated the etiologic characteristics of Shigella (S.) sonnei strains causing outbreaks and sporadic cases in some areas of Guangdong province and Guangxi Zhuang Autonomous Region during 2014-2016. Methods: Fourteen S. sonnei strains isolated from outbreaks and 6 S. sonnei strains from sporadic cases from Guangdong and Liuzhou of Guangxi Zhuang Autonomous Region were tested for antimicrobial resistance and analyzed by pulsed-field gel electrophoresis (PFGE). Six typical strains were selected for whole genome sequencing typing and compared with 51 strains isolated both at home and abroad from NCBI genome database. Results: The antibiotic resistance test indicated the isolates had high resistance rate to ampicillin, tetracycline, gentamicin, trimethoprim/sulfamethoxazole and nalidixic acid, while sensitive to azithromycin, chloromycetin and imipenem. PFGE showed high similarity (93.2%) among the strains isolated from different areas. The whole genome sequencing analysis also revealed that all the typical strains were clustered into a same evolution branch, close to some strains from Korea. Conclusions: The S. sonnei strains isolated from some areas of Guangdong and Guangxi Zhuang Autonomous Region showed high resistance to commonly used antibiotics, but they were sensitive to azithromycin, chloramphenicol and imipenem. The isolates in this study also showed similar PFGE patterns and close phylogenic evolution.

Clinical investigation of early post-traumatic temporomandibular joint ankylosis and the role of repositioning discs in treatment.

This study investigated the development of temporomandibular joint (TMJ) ankylosis after condylar fracture and the functional results of surgery that included repositioning of the articular discs. In a total of 18 patients, there were 13 cases of fibrous ankylosis (type I) and 11 of partial bony ankylosis (type II). CT scans for both groups and MRI scans for type I patients were analysed. Intraoperative inspection of the damaged disc, the sites of adhesion or bony fusion, and remaining intra-articular movement was recorded. After release arthroplasty and repositioning of discs, follow-up was for 1 to 3.5 years (mean 2.2 years). Post-traumatic TMJ ankylosis was highly associated with sagittal and comminuted condylar fractures. Type I ankylosis usually formed in the 4th to 5th month post-trauma with mean interincisal opening distance of 18.3+/-5.5mm. Progression from type I to II ankylosis occurred 1 year post-trauma and caused a reduction of 5mm in the range of mouth opening. The disc was displaced for each of the involved joints, and intra-articular adhesions or ossification initiated at the site where there was no intervening disc present. After surgical repositioning of the disc, stable joint function and mouth opening from 30 to 45 mm were obtained in all patients but one (recurrence due to dislocation). Sagittal and comminuted condylar fractures predispose the TMJ to ankylosis, and the displacement of the articular disc plays a critical role. Early surgical intervention to reposition the disc was successful for early trauma-induced TMJ ankylosis.

[Study on Salmonella serotyping by use of Microsphere-based Liquid Array method].

OBJECTIVE: To understand the effect of serotyping on Salmonella isolates, by use of Microsphere-based Liquid Array method, among diarrhea patients, in Guangdong. METHODS: Salmonella isolated from humans in Guangdong province were serotyped on the Microsphere-based Liquid Array platform with SSA kit. RESULTS: A total of 4 942 Salmonella strains with 189 serotypes, were identified in Guangdong province in 2010-2014. The top 100 serotypes accounted for 98.08% (4 847/4 942) of all the strains. 98% of the top 100 species serotypes could completely be serotyped with SSA kit. In order to detect O antigen among 198 isolates with SSA kit, 181 strains were carrying the O antigen, with the coincidence rate as 100%. However, under the SSA, 98.32% (528/537) of the H antigen could be detected and were consistent with the traditional serum agglutination test. The coincidence rate of fljB gene was 93.09% (175/188), with false negative rate and false positive rate of fljB gene as 7.35% (9/134) and 7.41% (4/54) respectively. The coincidence rate of sdf gene and Vi gene were 100%. 11 out of the 12 Salmonella strains could not be serotyped under the traditional methods but were successfully serotyped by the molecular serotyping method. CONCLUSIONS: Using the SSA kit, more than 96% of the anthropogenic Salmonella strains could be serotyped in Guangdong province. Comparing with the traditional methods, the coincidence rate of serotyping appeared over 98% . Under the Microsphere-based Liquid Array techniques, the molecular serotyping method appeared faster and more accurate on Salmonella serotyping than those traditional methods.

[Active etiological surveillance for foodborne diseases in Guangdong province, 2013-2014].

Objective: To study the infection status, serotypes, drug resistance and molecular characteristics of Salmonella, Shigella, Vibrio parahemolyticus, enterotoxigenic Escherichia (E.) coli (ETEC), pathogenic E. coli (EPEC), Shiga Toxin producing E. coli (STEC) and Enteroinvasive E. coli (EIEC) collected from diarrhea patients in Guangdong. Methods: The strains of Salmonella, Shigella, V. parahemolyticus and 4 kinds of E. coli isolated from foodborne diseases surveillance during 2013-2014 were collected to conduct serotyping, drug resistance test and pulsed-field gel electrophoresis (PFGE). Results: A total of 3 372 stains of pathogens were isolated from 57 834 stool samples during 2013-2014, the overall positive rate was 5.83% and the positive rate of Salmonella was highest, followed by that of V. parahemolyticus, 4 kinds of E. coli and Shigella. And 3 213 strains of Salmonella were divided into 143 serotypes. The most prevalent serotypes were Salmonella typhimurium, 4, 5, 12: i:-, Enteritidis, Stanley and Derby. Salmonella was sensitive to cephalosporin and fluoroquinolones, and showed significant differences in drug resistance rate among different serotypes. In top 10 common serotypes, S. enteritidis and S. derby were most resistant to cephalosporin and ciprofloxacin respectively. PFGE was performed for 2 289 strains of Salmonella, showing distribution diversity and significant fingerprint polymorphisms. The 85 strains of V. parahemolyticus were divided into 10 serotypes, O3∶K6 (61.18%) was the most common serotype, followed by O4∶K8. The results showed that the carrying rate of virulence genes tdh (81.18%) was high, while the carrying rate of trh was low (7.06%), and there were 10 strains carrying no the two kinds of virulence genes. The sensitive rate of V. parahemolyticus to imipenem, nalidixic acid, SMZ-TMP, chloramphenicol and tetracycline were more than 95%. Thirteen strains of Shigella were detected, including 9 strains of Shigella sonnei, 3 strains of Shigella flexneri and 1 strains of Shigella bogdii. The strains all showed sensitivity to ceftazidime, ciprofloxacin and chloramphenicol (76.92%). There were 86 strains of E. coli detected, including 29 strains of ETEC (33.72%), 27 strains of EPEC (31.39%), 27 strains of STEC (31.39%) and 3 strains of EIEC (3.48%). Conclusions: In the active etiological surveillance for foodborne diseases in Guangdong during 2013-2014, the detection rate of Salmonella was highest (5.57% ), followed by that of V. parahemolyticus, 4 kinds of E. coli and Shigella. Salmonella, V. parahemolyticus and Shigella were sensitive to cephalosporin and fluoroquinolones. Clustered cases of Salmonella infection were found in the surveillance, but no outbreaks occurred.

Correlation between the disc status in MRI and the different types of traumatic temporomandibular joint ankylosis.

OBJECTIVES: We aimed to investigate the correlation between the disc status in MRI and the different types of traumatic temporomandibular joint (TMJ) ankylosis. METHODS: 51 consecutive patients (69 joints), diagnosed with traumatic TMJ ankylosis with a residual condyle (Types A2 and A3), were included in this study. All patients had pre-operative MRI, which was reviewed to determine the disc shape, length and position. The results were compared using the Mann-Whitney test. RESULTS: There were 37 joints of Type A2 ankylosis and 32 joints of Type A3. All joints of Type A2 and 27 joints of Type A3 (84.4%) definitely had a discernible disc, while 5 joints of Type A3 had no discernible discs. Among the discernible discs, the lateral disc of Type A2 and the whole disc of Type A3 had severe deformity, while the medial disc of Type A2 had mild deformity. The mean (standard deviation) disc length was 10.88 (1.19) mm in Type A2, but 7.50 (0.82) mm in Type A3. There was a significant difference between Types A2 and A3 (p < 0.05). As for the disc position, the intermediate position was found in all joints. CONCLUSIONS: There is a correlation between the disc status and the different types of traumatic TMJ ankylosis. Therefore, MRI examination is needed to help treatment planning and predict post-operative TMJ function.

Restoration of X-ray and etoposide resistance, Ku-end binding activity and V(D) J recombination to the Chinese hamster sxi-3 mutant by a hamster Ku86 cDNA.

Ku is a heterodimeric protein composed of 86 and 70 kDa subunits that binds preferentially to the double-stranded ends of DNA. Recent molecular characterization of ionizing-radiation sensitive (IRs) mutants belonging to the XRCC5 complementation group demonstrated the involvement of Ku in DNA double-strand break (DSB) repair and lymphoid V(D)J recombination. Here, we describe the isolation of a full-length hamster cDNA encoding the large subunit of the Ku heterodimer and demonstrate that the stable expression of this cDNA can functionally restore IR, Ku DNA end-binding activity and V(D)J recombination proficiency in the Chinese hamster IRs sxi-3 mutant. Moreover, we also demonstrate that sxi-3 cells are hypersensitive to etoposide, a DNA topoisomerase II inhibitor, and that resistance to this drug was restored by the Ku86 cDNA. These experiments suggest that a defect in the large subunit of the heterodimeric Ku protein is the sole factor responsible for the known defects of sxi-3 cells and our data of further support the role of Ku in DNA DSB repair and V(D)J recombination.