The poly(ADP-ribose) polymerases (PARPs) is an important group of enzymes in DNA repair pathways, especially the base excision repair (BER) for DNA single-strand breaks (SSBs) repair. Inhibition of PARP in DNA repair-defective tumors (like those with BRAC1/2 mutations) can lead to cell death and genomic instability, what is so called "synthetic lethality". Currently, PARP inhibitors combined with cytotoxic chemotherapeutic agents in the treatment of BRCA-1/2 deficient cancers are in the clinical development. In this review, we will be focused on the development of combination application of PARP inhibitors with other anticancer agents in clinical trials.

Objective To investigate the adaptive immune responses in elderly patients with chronic obstructive pulmonary disease during acute exacerbations (ECOPD) and effects of the immunostimulating agent Pidotimod in ECOPD patients. Methods A randomized, prospective clinical trial was held, and 103 patients with ECOPD were recruited into the study. Seventy-five patients aged 65 years and over were divided into two groups: 38 patients with general treatment as a control group and 37 patients with general treatment plus pidotimod as an experimental group. Another non-elderly groups comprised 28 patients younger than 65, and 20 healthy individuals served as the healthy elderly control. Levels of CD3+ , CD4+ , CD8+ , CD4+ /CD8+ in peripheral blood were measured by flow cytometry at baseline (the 1st day) and at the 15th and 30th treatment day, meanwhile, the clinical conditions were evaluated. Results Ninety-one patients completed the trial (32 in experimental group,34 in control group and 25 in non-eldely group). The experimental group and control group were statistically homogeneous. The aged COPD intervention group and aged COPD control had a more decreased CD4+ level, CD4+/CD8+ ratio and more increased CD8+ level, while compared with aged health control and non-elderly COPD control (all P

Objective To determine the safety of the fetal olfactory ensheathing cell(OEC) transplantation in patients with chronic spinal cord injury (SCI) by examination of the magnetic resonance imaging (MRI). Methods A prospective clinical study involving 16 patients with chronic SCI was designed to investigate the feasibility and biological safety of the fetal OEC transplantation in treatment of SCI. The olfactory bulbs from the 3-4-month-old aborted human fetuses following the strict ethical guidelines were harvested and trypsinized down to single fetal OEC. These cells were then cultured for 12-17 days and were prepared for a clinical use. From November 2001 to December 2002, 16 patients with chronic SCI were randomly enrolled. The patients suffered from SCI for 1.5-8 years (average 4.3 years) after the injury. The suspension (50 μl) containing about 1×106 fetal OECs was transplanted by an injection into the patients' spinal cords above and below the injury site. All the patients were assessed before the transplantation and were followed up with MRI for 29-42 months (average 38 mon) after the transplantation. Results No cell-related adverse effects were observed in any patient during the follow-up period. The follow-up with MRI did not reveal any development of optic glial tumor, tumor-like mass, new hemorrhage, edema, expanding cyst, new cyst formation, infection or disruption of the neural structure in the transplant site of all the patients. Conclusion This is the first clinical study demonstrating the long-term safety of the OEC therapy for SCI. The results indicate that our protocol is feasible and safe in treatment of patients with chronic SCI within 38 months after the injury. Although the size of the samples for our study was not big enough, the positive results of the study have encouraged us to make a further research in this field.

Objective:To investigate the clinical characteristics and gene variation of asparagine synthase deficiency that is caused by ASNS gene variation. Methods:In Department of Neuroendocrine Pediatrics, Affiliated Hospital of Qingdao University from October 2018 to February 2020, the clinical data of a family of asparagine synthase deficiency were analyzed retrospectively.The pathogenic mutation of the proband was screened by the full exon analysis technique.The pathogenic sites of candidate genes were determined by combining the phenotype of the proband.In the heterotopic spot of the proband, his parents and other family members were verified by Sanger sequencing.Meanwhile, the relevant literature database was consulted, and the reported ASNS mutation related cases were collected and reviewed. Results:The female with proband visited the hospital at the age of 4 months, and she had recurrent convulsions at the age of about 3 months.Physical examination showed that the child suffered from microcephaly, and mental and motor retardation.Meanwhile, video electroencephalogram examination displayed extensive moderate high amplitude spiny slow wave and sharp slow wave.Exon sequencing illustrated that the compound heterozygous variants of ASNS gene were c. 1211G>A (p.R404H) and c. 1643C>T (p.S548F), respectively.c.1211G>A was a known pathogenic variant, and c. 1643C>T was a new variant.The proband′s younger brother visited the hospital at the age of 2 months, developed convulsions at the age of 1 month, and developed mental and motor retardation.Electroencephalogram displayed that bilateral posterior head was dominant, multiple foci and extensive spike wave, and spike slow wave and fast wave were distributed.Sanger sequencing revealed the same ASNS compound heterozygous variants as the proband.Both of them died of status convulsion at the age of 7 months and 6 months, respectively. Conclusions:This study is helpful to further understand the clinical features of the disease and reveal a new pathogenic mutation of ASNS gene, so as to enrich the mutation spectrum of ASNS gene, thus providing important basis for clinical treatment and genetic counseling.

To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor β1 (TGF-β1) gene in vitro, cultured BMSCs were transfected with the complexes of pcDNA3-TGF-β1 and Lipofectamine Reagent in vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type Ⅰ propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type Ⅰ expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-β1 gene could promote the osteogenic potential of cultured BMSCs.

To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor β1 (TGF-β1) gene in vitro, cultured BMSCs were transfected with the complexes of pcDNA3-TGF-β1 and Lipofectamine Reagent in vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type Ⅰ propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type Ⅰ expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-β1 gene could promote the osteogenic potential of cultured BMSCs.

S and NO production induced by rhIL-1β in a concentration-dependent manner. It is suggested that PDTC can inhibit NO production and iNOS mRNA expression induced by IL-1β, which may provide an alternative method for the treatment of osteoarthritis.

OBJECTIVETo investigate the frequency of p16 and p15 gene methylation in multiple myeloma (MM), and its relationship with bone marrow cell apoptosis and clinical outcome.

METHODSTwenty-two patients with MM were studied to detect p16 and p15 gene methylation. Methylation-specific polymerase chain reaction (MSP) was used to detect gene methylation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) was used to detect cell apoptosis.

RESULTSp16 and/or p15 gene methylatoin was detected in 10 of 22 patients (45.4%). There were 3 patients with p16 gene methylation, 9 patients with p15 gene methylation, and 2 patients with both genes methylation. The incidence of methylation of p15 gene was higher than that of p16 gene (P < 0.05). The patients with p16 and/or p15 gene methylation had a delayed cell apoptosis, poor response to chemotherapy, and a short over-all survival (OS).

CONCLUSIONThe methylation of p16 and/or p15 gene plays a key role in MM apoptosis pathogenesis. The patients with both p16 and p15 gene methylation had a poor prognosis.

Apoptosis ; Cell Cycle Proteins ; Cyclin-Dependent Kinase Inhibitor p15 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Gene Silencing ; Genes, p16 ; Humans ; Multiple Myeloma ; genetics ; pathology ; Prognosis ; Transcription Factors ; genetics ; Tumor Suppressor Proteins

The feasibility of using gene therapy to treat full-thickness articular cartilage defects was investigated with respect to the transfection and expression of exogenous transforming growth factor (TGF)-beta 1 genes in bone marrow-derived mesenchymal stem cells (MSCs) in vitro. The full-length rat TGF-beta 1 cDNA was transfected to MSCs mediated by lipofectamine and then selected with G418, a synthetic neomycin analog. The transient and stable expression of TGF-beta 1 by MSCs was detected by using immunohistochemical staining. The lipofectamine-mediated gene therapy efficiently transfected MSCs in vitro with the TGF-beta 1 gene causing a marked up-regulation in TGF-beta 1 expression as compared with the vector-transfected control groups, and the increased expression persisted for at least 4 weeks after selected with G418. It was suggested that bone marrow-derived MSCs were susceptible to in vitro lipofectamine mediated TGF-beta 1 gene transfer and that transgene expression persisted for at least 4 weeks. Having successfully combined the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, i.e. molecular tissue engineering, are put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in research. Using this technique, we have a new powerful tool with which: (1) to modify the functional biology of articular tissue repair along defined pathways of growth and differentiation and (2) to affect a better repair of full-thickness articular cartilage defects that occur as a result of injury and osteoarthritis.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; cytology ; Gene Transfer Techniques ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; Stem Cells ; cytology ; metabolism ; Tissue Engineering ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics

BACKGROUND: It was thought that there was no regeneration capacityin central nerves. Recent research shows that regeneration capacity of injured neural axons and recovery of some neurological functions can be achieved by changing local surroundings after spinal cord injury (SCI).OBJECTIVE: To probe into whether the transplantation of fetal olfactory ensheathing cells (OECs) in recovering the neurological functions of patients with chronic SCI is safe, feasible, and effective.DESIGN: Auto-control observation before and after surgery.SETTING: Neurological Research and Treatment Center of Beijing Xishan Hospital; Second Department of Neurosurgery in Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences; Second Department of Neurosurgery in Naval General Hospital.PARTICIPANTS: A total of 171 patients with chronic spinal cord injury were selected from the Second Department of Neurosurgery in Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences and the Second Department of Neurosurgery in Naval General Hospital betweenNovember 2001 and February 2003, of which there are 147 patients with complete injury and 24 ones with incomplete injury. Post-injury period ranged from 0.5 to 18 years. Process of treatment is discussed and permitted by relevant Medical Ethics Committees. Cells were obtained from voluntary donors and patients agreed to receive the treatment.METHODS: ① Fetal olfactory bulbs were cultured for 12-17 days after being digested into single cells. ② Fetal OECs were transplanted into sites rostral and caudal to the epienter. ③ Neurological functions of all patients 2-8 weeks before and after operation were evaluated according to the scoring standard of ASIA.MAIN OUTCOME MEASURES: ① Status of functional recovery in spinal cord of patients after transplantation of OECs. ② Harmful events and side effects.RESULTS: A total of 171 patients were involved in the analysis of results.①Status of functional recovery in spinal cord of patients with OECs transplantation: Partial neurological functions of 171 patients rapidly recovered,whose motor function score increased from (34.5±20.3) points before operation to (42.0±20.0) points (P ＜ 0.001) after operation, score of light touch increased from (47.2±24.0) points to (61.8±23.0) points (P ＜ 0.001) after operation,score of pain sense increased from (48.6±23.5) points to (64.0±22.8) points (P ＜ 0.001). ②Harmful events and side-effects: Early manifestations of spinal cord injury induced by infection in surgical area of one patient aggravated; two patients suffered from serious pulmonary infection,one patient from thalamic hemorrhage. Three patients mentioned above died of serious respiration and circulatory failures.CONCLUSION: OEC transplantation can rapidly promote partial neurological function of patients with chronic SCI, while the mechanism needs further observing.