OBJECTIVETo investigate the correlation of the deleted azoospermia (DAZ) gene copy related to gr/gr and b2/b3 deletions in the AZFc region with male spermatogenic impairment.

METHODSThis study included 121 infertile men with different de- grees of spermatogenic impairment and 95 healthy donors from the sperm bank. Using PCR, PCR-RFLP, and Y chromosome specific sequence tagged sites (STS) , we analyzed the association of DAZ gene copy deletions related to gr/gr and b2/b3 deletions in the AZFc region with spermatogenic impairment.

RESULTSThere were 15 cases of gr/gr deletion (12. 40% ) and 6 cases of b2/b3 deletion (4.96%) in the infertility group as compared with 13 cases of gr/gr deletion (13.68%) and 1 case of b2/b3 deletion (1.05%) in the control. Analysis of the DAZ-specific single nucleotide variant (SNV) loci revealed 11 gr/gr-DAZI/DAZ2 deletions (9.09%), 4 gr/gr-DAZ3/DAZ4 deletions (3.31%), and 6 b2/b3-DAZ1/DAZ2 deletions (4.96%) in the infertile men in comparison with 3 gr/ gr-DAZ1/DAZ2 deletions (3.16%), 10 gr/gr-DAZ3/DAZ4 deletions (10.53%), and 1 b2/b3- DAZ3/DAZ4 deletion (1.05%) in the control.

CONCLUSIONPartial deletions of gr/gr and b2/b3 exist in both healthy men and male patients with different degrees of spermatogenic impairment and cannot be considered as a risk factor for spermatogenesis impairment. However, deletions of different DAZ duplicons in gr/gr and b2/b3 deletions have different effects on spermatogenesis. DAZ1/DAZ2 instead of DAZ3/DAZ4 deletions might be associated with spermatogenesis impairment.

Deleted in Azoospermia 1 Protein ; Gene Deletion ; Gene Dosage ; Humans ; Male ; RNA-Binding Proteins ; genetics ; Spermatogenesis ; genetics

Objective To develop a sensitive and specific microarray for detecting mutations of HBV pre-core/core and basic core promoter regions in the clinic. Methods Site-specific oligonucleotide probes were designed and immobilized to microarray slides and hybridized to HBV gene fragments amplified with specific biotin-labeled primer using asymmetrical PCR. The specificity and sensitivity of the method were estimated. And the microarray was applied to detect 138 clinical serum samples with HBV-DNA.Results The mutations of HBV pre-core/core and basic core promoter regions can be specifically detected using the microarray, and the sensitivity was 1 × 101 copies/μl. Among 138 samples,40 samples had T1762/A1764 mutation, 1 lsamples had C1814 mutation, and 16 samples had A1896 mutation. The A1896 mutation rate in high HBV-DNA load group was significantly higher than that in low HBV-DNA load group (P ＜ 0. 01 ). Conclusion An DNA microarray assay was successfully established to detect the mutations in HBV pre-core/core and basic core promoter regions. The A1896 mutation in Pre-core/core region maybe involve in duplication of HBV.

Objective To optimize the preparation of nanoparticles (NP) encapsulating antisense oligodeoxynucleotides (ASODN) and investigate the effects on inhibition of C6 glioma cells. Methods ASODN coated in NT were prepared by interfacial polymerization of butyleyanoacrylate (BCA). Inverted microscope was used to observe the viability of C6 cells transfected by free ASODN, ASODN in NP, ASODN-NP (ASODN sticking to NP) and BCA-NP, respectively. Cell cycle of C6 cells was studied by flow cytometry (FCM), and CCK-8 assay was performed to examine the cytotoxicity and proliferation of C6 cells. Results Compared with the control group, all groups, except BCA-NP group, after transfection with NPs appeared cell morphological changes; C6 cells were detached from the matrix, the cell density was reduced and the cell viability was poor; ASODN in NP group was most significant in a time-dependent manner. The cell cycle in ASODN-in-NP group varied obviously compared with the BCA-NP group, and the number of the cells in the GO/GI phase was increased and the cell number in S phase was decreased significantly (P<0.05). The results of CCK-8 assay showed that all groups, but BCA-NP group, produced the inhibition of the cell proliferation to different degrees, and the inhibitory effect was increased with the final concentration increment, especially remarkably in ASODN-in-NP group (P<0.05). Conclusion ASODN in NP can inhibit the proliferation and cause cell cycle changes of C6 cells effectively after transfected with ASODN in NP, exerting significantly growth inhibitory effect on C6 glioma cells.

OBJECTIVETo study the impact of neoadjuvant therapy on lymph nodes retrieval in locally advanced mid-low rectal carcinoma.

METHODSData collected from 120 patients with locally advanced mid-low rectal cancer (T2-4 and/or N1-2M0) treated from January 2005 to June 2008 was investigated. The patients were divided into two groups: the study group (n=54) was treated with neoadjuvant therapy (preoperative radiation with a total dosage of 50 Gy and synchronous 5-Fu-based chemotherapy) followed by radical tumor resection 4-6 weeks after;the control group (n=66) underwent primary surgery without neoadjuvant therapy. The clinical stage was evaluated before and after neoadjuvant therapy. The total lymph nodes yields, as well as the tumor-positive lymph nodes of each resected specimen was compared between the two groups statistically.

RESULTSClinical downstage was achieved in 30 cases (56%) in study group after neoadjuvant therapy. The number of total lymph nodes and positive lymph nodes harvested from each resected specimen in the control group were 14+/-7 and 2.2+/-3.7, meanwhile those were 9+/-6 and 0.7+/-2.4 in study group, which were all significantly lower than those in control group (P<0.01).

CONCLUSIONSPreoperative radiotherapy combined with chemotherapy can downstage the tumor and reduce the retrieval rate of total lymph nodes and positive lymph nodes in locally advanced rectal cancer. It is necessary to retrieve as many lymph nodes as possible for it has some prognostic significance for the patients.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Prognosis ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the efficacy of neoadjuvant radiotherapy alone versus chemoradiotherapy in patients with mid-low locally advanced rectal cancer.

METHODSData of 69 patients with advanced (stage T(3) or T(4)) rectal cancer, undergone neoadjuvant therapy in our hospital from October 1997 to October 2007, were analyzed retrospectively. Forty patients received preoperative radiotherapy (50 Gy in 25 fractions over 5 weeks) alone (RT group), and 29 patients received preoperative radiotherapy concomitant with 5-FU/leucovorin -based preoperative chemoradiotherapy (CRT group). Radical surgery was performed 4-6 weeks after radiation therapy by the rule of TME.

RESULTSAll the patients underwent operations, including 26 abdominoperineal resections, 27 anterior resections, 10 Parks operations and 6 Hartmann's procedures. The sphincter preservation rate was 47.5%(19/40) in RT group, and 62.1%(18/29) in CRT group(P>0.05). In pathological findings, tumor and nodal downstaging were observed in 12 patients of RT group (30.0%), and 17 of CRT group (58.6%)(P<0.05). In RT group, 3 patients (7.5%) showed pathological complete regression (pCR), and the overall response rate (CR plus PR) was 60%(24/40). In CRT group, 4(13.8%) showed pCR and the overall response rate was 79.3%(23/29). There was significant difference of the overall response rate between two groups. Three-year disease-free survival for all patients was 77.3%.

CONCLUSIONFor patients with locally advanced rectal cancer, neoadjuvant chemoradiotherapy provides higher sphincter preservation rate, overall response rate and better down-staging as compared to radiotherapy alone.

Chemotherapy, Adjuvant ; Female ; Humans ; Male ; Middle Aged ; Neoadjuvant Therapy ; methods ; Neoplasm Staging ; Radiotherapy, Adjuvant ; methods ; Rectal Neoplasms ; pathology ; radiotherapy ; therapy ; Retrospective Studies

Objective To explore the effect of aspirin(ASP) combined with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) inducer TIC10 on apoptosis and autophagy of cervical cancer Siha and HeLa cells. Methods HPV16 positive squamous carcinoma Siha cells and HPV18 positive cervical adenocarcinoma HeLa cells were selected as research objects. Two cell lines were treated with ASP,TIC10 or two drugs combination. The effects of ASP and TIC10 on cell proliferation were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay,cell migration ability was detected by cell scratch assay,cell clone formation ability was detected by plate clone formation assay,and the levels of cleaved Caspase-3/8,LC3A/B,Beclin1 were detected by Western Blot. Results The MTT showed that the proliferations of Siha and HeLa cells were inhibited by APS in a concentration dependent manner (all P < 0. 01). After treated with TIC10 combined with ASP for 48 h,the cell proliferation inhibition rates of Siha and HeLa cells were higher than thosetreated with TIC10 alone (all P < 0. 01). After treated with TIC10 or ASP for 24 h, the migration rates of Siha and He- La cells in the experimental groups [ASP: (19. 61 ± 1. 17) ％,(23. 75 ± 0. 78) ％; TIC10: (16. 89 ± 1. 47) ％,(20. 59 ± 2. 01) ％]were lower than those of control group [(41. 18 ± 2. 01) ％,(40. 83 ± 3. 77) ％],all P < 0. 01. After two weeks of treatmentof ASP and TIC10,the CFE(colony-forming efficiency) rates of Siha and HeLa cells in the experimental groups [ASP: (24. 93 ± 2. 12) ％, (26. 47 ± 3. 30) ％, TIC10: (17. 33 ± 1. 50) ％,(19. 13 ± 4. 99) ％]were lower than those of control group [(69. 60 ± 3. 54) ％,(68. 40 ± 4. 20) ％],all P < 0. 01. Western blot assays showed that the expression of cleaved Caspase-3/8 and autophagy related protein LC3A/B,Beclin1 were higher than those of control group (all P < 0. 01). Conclusion ASP increases cervical cancer cells apoptosis with TRAIL inducer TIC10 through autophagy enhancement.

Objective:To investigate the association of R wave peak time(RWPT)with severity of coronary artery disease and short-term prognosis in patients with acute coronary syndrome(ACS).Methods:A total of 133 ACS patients who were treated in Shenzhen Longhua District Central Hospital between January 2019 and June 2022 and received coronary angiography were selected.According to number of diseased coronary arteries,they were divided into single-vessel coronary disease group(n=43),double-vessel coronary disease group(n=51)and multi-vessel coronary disease group(n=39);according to severity of coronary artery disease,they were divided into mild group(n=40),medium group(n=48)and severe group(n=45).ECG indexes were compared among groups with different number of diseased coronary arteries and severity of coronary artery disease.Pearson correlation analysis was used to analyze corre-lation between ECG indexes and Gensini score;patients were divided into major adverse cardiovascular events(MACE)group(n=46)and no MACE group(n=87)according to presence of MACE during hospitalization.Clinical data were compared between two groups,and multivariate Logistic regression was used to analyze risk factors of MACE during hospi-talization in ACS patients.Results:QRS duration and RWPT significantly increased according to the order of single-ves-sel,double-vessel and multi-vessel coronary disease group(P＜0.05 or＜0.01);QRS duration and RWPT significantly increased according to the order of mild,medium and severe group(P＜0.05 or＜0.01).Pearson correlation analysis indi-cated that QRS duration and RWPT were significant positively correlated with Gensini score in ACS patients(r=0.222,0.557,P=0.010,＜0.001).Multivariate Logistic regression analysis indicated that hypertension,diabetes,RWPT,Gensini score and multi-vessel disease were independent risk factors for MACE during hospitalization in ACS patients(OR=3.171～9.360,P＜0.05 or＜0.01).Conclusion:RWPT is closely related to the severity of coronary artery disease in ACS patients.RWPT detection helps to guide risk stratification and secondary prevention of ACS patients,therefore im-prove their short-term prognosis.

OBJECTIVETo investigate the post-operative complications of aortic endovascular grafting exclusion (EVGE) and its reasons and treatments.

METHODSClinical data of 82 cases received aortic endovascular grafting exclusion from January 2002 to October 2008 were retrospectively analyzed. Seventy-one cases were male and 11 cases were female with the age of 33 to 78 years and the average age of 49.2 years. There were 66 cases of thoracic aortic dissecting aneurysms and 16 cases of abdominal aortic aneurysm. The effect, post-operational complications and its treatment were investigated.

RESULTSThere were 90.1% patients had been followed up with the time of 3 to 78 months with technical success of 90.3%, clinical success of 94.1%, peri-operational mortality of 2.4%, total mortality of 6.1% and mortality associated with EVGE of 2.4%. Twenty-one cases underwent complications including type I endoleak (13 cases), abdominal aortoduodenal fistula (1 case), narrow true lumen (2 cases), reverse Stanford A dissection (2 cases), post EVGE syndrome (12 cases), delayed healing of inguinal incision (5 cases), constipation (3 cases), cerebral infarction (1 case). No paraplegia, left subclavian artery ischemia, contrast media associated nephrosis, ischemic colitis, ischemic neurologic injury, and artery embolism occurred. Post operation 4 cases had the second intervention including 2 type I endoleak and 2 narrow true lumen.

CONCLUSIONSThe technique-related complications still hinder the long-term effect of EVGE. It needs to be further investigated on technique improvement and treatment standardization.

Adult ; Aged ; Aneurysm, Dissecting ; surgery ; Aortic Aneurysm ; surgery ; Blood Vessel Prosthesis Implantation ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; therapy ; Retrospective Studies

BACKGROUNDThe long term prognosis of unprotected left main coronary artery (LMCA) stenting is controversial. This study was conducted to evaluate the immediate and long term outcomes of LMCA stenting in Chinese patients and to determine which factors affect the outcomes.

METHODSFrom May 1997 to March 2003, 224 patients in 23 hospitals underwent elective unprotected LMCA stenting with bare metal stents. Their clinical records were analysed to ascertain immediate and long term outcomes of LMCA stenting as well as factors influencing the prognosis.

RESULTSStents were implanted into LMCA successfully in 223 cases (99.6 %). One death (0.5%) and one case of non-Q wave nonfatal myocardial infarction (MI) occurred in hospital. The mean follow-up time was (15.6 +/- 12.3) months. Cardiac death developed in 10 cases (4.5%), noncardiac death in 2 cases (0.9%), nonfatal MI in 4 cases (1.8%), target lesion revascularization (TLR) of LMCA in 26 cases (11.7%) and TLR of nonLMCA in 37 cases (16.5%). Univariate analysis showed that cardiac death correlated with left ventricular ejection fraction (LVEF < 40%), female gender and LMCA combined with multivessel disease; that major adverse cardiac events (MACE) correlated with LVEF < 40%, bifurcation lesion and incomplete revascularization. Logistic regression analysis revealed that LVEF < 40% and female gender were independent predictors of cardiac death and MACE. Follow-up angiography was performed in 102 cases (45.7%). The restenosis rate was 31.4%.

CONCLUSIONSLong-term outcomes of stenting for selected patients with unprotected LMCA stenosis is acceptable. It should be performed in inoperable or low risk patients with LVEF > or = 40% and isolated LMCA disease or LMCA combined with multivessel diseases in whom complete revascularization can be obtained.

Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Disease ; therapy ; Coronary Restenosis ; etiology ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stents ; adverse effects ; Treatment Outcome

DNA genetic markers have always played important roles in individual identification, kinship analysis, ancestry inference and phenotype characterization in the field of forensic medicine. DNA methylation has unique advantages in biological age inference, body fluid identification and prediction of phenotypes. The majority of current studies independently examine DNA and DNA methylation markers using various workflows, and they use various analytical procedures to interpret the biological information these two markers present. Integrated methods detect DNA and DNA methylation markers simultaneously through a single experimental workflow using the same preparation of sample. Therefore, they can effectively reduce consumption of time and cost, streamline experimental procedures, and preserve valuable DNA samples taken from crime scenes. In this paper, the integrated detection approaches of DNA and DNA methylation markers on different detection platforms were reviewed. In order to convert methylation modifications to detectable forms, several options were available for pretreatment of genomic DNA, including digestion with methylation-sensitive restriction enzyme, affinity enrichment of methylated fragments, conversion of methylated or unmethylated cytosine. Multiplexed primers can be designed for DNA markers and converted DNA methylation markers for co-amplification. The schemes of using capillary electrophoresis platform for integrated detection add the pretreatment of genomic DNA on the basis of detecting DNA genetic markers. DNA and DNA methylation markers are then integrated by co-amplification. But the limited number of fluorescent options available and the length of amplicons restrict the type and quantity of markers that can be integrated into a panel. Pyrophosphate sequencing also supports integrated detection of DNA and DNA methylation markers. On this platform, due to the conversion of unmethylated cytosine to thymine after treatment with bisulfite, the methylation level of CpG site can be directly calculated using the peak height ratio of cytosine bases and thymine bases. Therefore, the methylation levels and SNP typing can be simultaneously obtained. However, due to the limited read length of sequencing, the detection of markers with longer amplicons is restricted. It is not conducive to fully interpret the complete information of the target sequence. Next-generation sequencing also supports integrated detection of DNA and DNA methylation markers. A preliminary experimental process including DNA extraction, pretreatment of genomic DNA, co-preparation of DNA and DNA methylation library and co-sequencing, has been formed based on the next-generation sequencing platform. It confirmed the feasibility of next-generation sequencing technology for integrated detection of DNA and DNA methylation markers. In field of biomedicine, various integrated detection schemes and corresponding data analysis approaches of DNA and DNA genetic markers developed based on the above detection process.Co-analysis can simultaneously obtain the genomic genetic and epigenetic information through a single analytic process. These schemes suggest that next-generation sequencing may be an effective method for achieving more accurate and highly integrated detection, helping to explore the potential for application in forensic biological samples. We finally explore the impact of interactions between sites and different pretreatment methods on the integrated detection of DNA and DNA methylation markers, and also propose the challenge of applying third-generation sequencing for integrated detection in forensic samples.