[Epidemiological distribution of mosaic loss of chromosome Y in adult men in 10 areas in China and its prospective association with lung cancer].

Objective: To detect the prevalence of mosaic loss of chromosome Y in adult men in ten study areas in China, describe the epidemiological distribution of mosaic loss of chromosome Y (mLOY) carriers and assess its prospective association with lung cancer. Methods: Based on the data from baseline survey, genetic analysis and follow-up (as of December 31, 2018) from China Kadoorie Biobank, we used Mosaic Chromosomal Alterations pipeline to detect mLOY carriers in 10 areas in China and described the epidemiological characteristics of mLOY carriers in adult men, including age, area distribution, lifestyle and disease history. We used multivariate logistic regression model to identify the potential relevant factor of mLOY. Cox proportional hazard regression model was fitted to assess the prospective association of mLOY with lung cancer. Stratification analysis were conducted to evaluate the potential modification effects of smoking and age. We also conducted mediation analysis to assess the mediating effect of mLOY in the association between smoking and lung cancer. Results: A total of 42 859 adult men were included in our analysis, in whom 2 458 mLOY carriers were detected (5.7%). The detection rate increased with age (P<0.05). The detection rate was higher in urban area (7.3%±0.2%) than that in rural area (4.7%±0.1%). The results of logistic regression analysis indicated that smoking might be a risk factor for the detection of mLOY (OR=1.49, 95%CI:1.36-1.64). After follow-up for average 11.1 years, 1 041 lung cancer cases were observed. The prospective analysis showed that mLOY carriers had an increased risk for lung cancer by 24% compared with non-mLOY carriers (HR=1.24, 95%CI:1.01-1.52) and expanded mLOY carriers (mLOY cell proportion ≥10%) had an increased risk for lung cancer by 50% (HR=1.50, 95%CI:1.13-2.00). Stratification analysis showed no modification effects of smoking and age in the association between mLOY and lung cancer (interaction P>0.05). Mediation analysis showed that mLOY could be a mediating factor in the association between smoking and lung cancer, the estimated effect was 0.09 (0.01-0.17). Conclusions: There were significant differences in the detection rate of mLOY in adult men with different social-economic characteristics and lifestyles in ten areas in China. Besides, mLOY carriers, especially expanded mLOY carriers, had increased risk for lung cancer and mLOY might be a mediating factor in the association between smoking and lung cancer.

[Clinical study of modified Shirodkar transvaginal cervical cerclage during pregnancy in the treatment of cervical insufficiency].

Objective: To investigate the clinical efficacy of modified Shirodkar transvaginal cervical cerclage (TVCC) in the treatment of cervical insufficiency (CI) and its impact on maternal and fetal outcomes. Methods: The clinical data of 218 pregnant women with CI admitted to Fu Xing Hospital, Capital Medical University from January 1, 2015 to August 31, 2021 was retrospectively analyzed. According to different surgical approaches, they were divided into modified Shirodkar TVCC treatment during pregnancy (TVCC group, 108 cases) and non-pregnant women underwent laparoscopic cervical cerclage (LACC) treatment (LACC group, 110 cases). The clinical data and pregnancy outcomes of the two groups were compared. Furthermore, the two groups of pregnant women were stratified according to cervical length (CL) to explore the effects of the two surgical methods on the pregnancy outcomes of CI women with different CL. Results: (1) Related indicators before and during cerclage: there were no complications such as massive hemorrhage, bladder injury and anesthesia accident in the two groups of pregnant women during cerclage. Compared with the LACC group, TVCC group had longer preoperative CL [(2.3±0.6) vs (2.7±0.6) cm], more intraoperative blood loss [(7.5±0.5) vs (14.4±1.4) ml] and longer hospital stay [(6.0±0.1) vs (7.3±0.4) day]. However, the operation time was shorter [(42.9±1.6) vs (25.9±1.4) minute] and the hospitalization cost was less [(9 912±120) vs (5 598±140) yuan], and the differences were statistically significant (all P<0.05). (2) Pregnancy outcomes: live birth rates were 95.4% (103/108) in the TVCC group and 96.4% (106/110) in the LACC group, showing no significant difference between the two groups (χ2=2.211, P=0.232). The preterm birth rate (12.0%, 13/108) in the TVCC group was higher than that in the LACC group (7.3%, 8/110), the neonatal birth weight was lower than that in the LACC group [(3 006±96) vs (3 225±42) g], and the proportion of low birth weight infants was higher than that in the LACC group [15.5% (16/103) vs 1.9% (2/106)], and the differences were statistically significant (all P<0.05). (3) Stratified analysis of CL: for pregnant women with CL<2.0 cm, the miscarriage rate of the TVCC group was higher than that of the LACC group (2/9 vs 3.0%), and the live birth rate was lower than that of the LACC group (7/9 vs 97.0%), and the differences were statistically significant (all P<0.05). For CL 2.0-<2.5 cm, 2.5-<3.0 cm, CL≥3.0 cm, there were no statistically significant differences in preterm birth rate and live birth rate between the two groups (all P>0.05). Conclusions: Modified Shirodkar TVCC is simple and easy to operate, which significantly reduces the cesarean section rate and medical cost compared with LACC, and there is no significant difference in the live birth rate. When there is inevitable late abortion, laparoscopic cerclage removal does not need to be performed again, which could reduce the second operation and is worthy of clinical application.

[Clinical efficacy of single/double 125I-seed strands combined with biliary stents in the treatment of malignant obstructive jaundice].

Objective: To compare the clinical efficacy of single/double 125I-seed strands combined with biliary stents in the treatment of malignant obstructive jaundice. Methods: Totally 67 cases of patients with malignant obstructive jaundice who received single/double125I-seed strands combined with biliary stents implantation from September 2018 to December 2021 were analyzed retrospectively. Among them, 36 patients received single 125I-seed strands combined with biliary stents (single strand group) and 31 patients received double 125I-seed strands combined with biliary stents(double strands group). The technical success rate, clinical success rate, complications, biochemical and tumor indexes at 8 weeks after operation [total bilirubin (TB), direct bilirubin (DB), alanine transaminase (ALT), aspartate transaminase (AST), carbohydrate antigen 19-9 (CA19-9)], stent patency time (SP), median progression-free survival time (mPFS) and median survival time (mOS) were analyzed. Results: There was no significant difference (P>0.05) in technical success rate (100% vs 100%), clinical success rate (97.2% vs 96.8%) and major complications (5.6% vs 6.5%) between single strand group and double strands group. There were significant differences in TB, DB, ALT, AST and CA19-9 indicators between the two groups before and 8 weeks after operation (all P<0.05), but there was no significant difference in the difference value of preoperative and postoperative 8-week indicators between the two groups (all P>0.05).The SP and mPFS of double-stranded stents were longer than those of single-stranded stents.[8.6 months (95%CI:6.9-10.4) vs 6.2 months (95%CI:5.8-6.6), 3.2 months (95%CI:3.0-3.4) vs 3.0 months (95%CI:2.9-3.1), all P<0.05]. The mOS of single and double strands groups was 11.2 months (95%CI:8.3-14.1) and 13.4 months (95%CI:9.9-16.9) respectively, with no statistical difference (P=0.137). Conclusion: Compared with single 125I-seed strands, double 125I-seed strands can prolong biliary SP and mPFS, but the long-term survival index still needs further observation.

[Epidemiological distribution characteristics of peripheral blood mosaic chromosomal alteration in adults from 10 regions of China].

Objective: To describe the epidemiological distribution characteristics of peripheral blood mosaic chromosomal alteration (mCA) in community adults aged 30-79 years in 10 regions of China. Methods: A total of 100 297 participants with complete baseline information (demographic characteristics, lifestyle, physical examination, etc.) and genotyping data of blood-derived DNA in ten regions of the China Kadoorie Biobank study were included. The mCAs were detected with the Mosaic Chromosomal Alterations pipeline, and logistic regression models were used to compare the differences in the detection rate of mCAs in different regions and populations. Results: A total of 5 810 mCA carriers were detected, with the detection rate of 5.8%. The standardized detection rate was 5.1%. The baseline detection rate of mCA increased with age, which were 3.4%, 5.0%, and 9.4% in those aged 30-, 51-, and >60 years, respectively (trend test P<0.001). A more significant proportion of mCAs were found in men (8.0%) than women (4.0%), as well as in urban areas (6.4%) than in rural areas (5.3%), the difference was significant (P<0.001). After adjusting for age and gender, the detection rate of mCA was higher in current smokers or people quitting smoking due to illness and people with low physical activity level, and the mCA detection rate was lower in obesy people (5.3%) than that in people with normal body weight (5.9%) (P=0.006). Conclusions: The detection rate of mCAs varied with region and population in community adults aged 30-79 years in 10 regions of China. The study results might contribute to the molecular identification of aging populations and guide precision prevention of age-related diseases such as cancers.

[Progress on genome-wide association studies on mosaic chromosomal alterations].

Mosaic chromosomal alteration (mCA) is referred to as large-scale somatic mutations on chromosomes, which results in diverse karyotypes in body. The mCA is regarded as one of the phenotypes of aging. Studies have revealed its associations with many chronic diseases such as hematopoietic cancers and cardiovascular diseases, but its genetic basis (e.g. genetic susceptibility variants) is still under-investigated. This paper reviews GWAS studies for mCA on autosomal chromosomes and sex chromosomes [mosaic loss of the Y chromosome (mLOY) and mosaic loss of the X chromosome (mLOX)] based on large population, respectively. Most of the genetic susceptibility loci found in studies for autosomal mCA were associated with copy-neutral loss of heterozygosity. The study of sex chromosome mCA focused on mosaic loss mutations. The number of genetic susceptibility loci for mLOY was high (up to 156), but it was relatively less for mLOX.

[Immune function and research progress of pentraxin 3 in pulmonary infectious diseases].

Pentraxins3 (PTX3) is an acute-phase protein of the pentraxin family that is synthetized and stored in a variety of cells. As an important mediator of innate immunity, PTX3 is rapidly released during microbial invasion and inflammatory response. It promotes the recognition of pathogens by myeloid cells through regulating complement activation. Recent studies have indicated that PTX3 concentrations in peripheral blood or tissues increase rapidly after infection, and the increased level is associated with the severity of the disease. Thus, PTX3 appears to be a vital clinical biomarker in the diagnosis and prognosis of pulmonary infectious diseases.

[Research progress on the epidemiological distribution and influencing factors of autosomal mosaic chromosomal alteration].

Somatic mosaicism is defined as the occurrence and accumulation of somatic mutations in humans, and mosaic chromosomal alterations (mCA) are recognized as one of the aging phenotypes due to their impact on genome integrity. With the coming acceleration of global population aging, understanding the prevalence and influencing factors of mCA will help to explore the "genomic instability" of human aging and its biological mechanisms and provide the scientific basis for the primary prevention of age-related diseases. This review aims to summarize the epidemiological distribution and influencing factors of autosomal mCA in peripheral blood based on previous large-scale population-based studies.

[Review of genome-wide association research of aging phenotypes].

"Active health" has been emphasized in "Healthy China 2030" in dealing with the challenges of population aging, so the anti-aging strategies are requires to be more precise and effective at both individual and population levels. Aging is influenced by both genetic and environmental factors. In the recent 20 years, the research of genetics of human ageing has been greatly facilitated owning to the development of high-throughput sequencing techniques, statistical methodology for multi-omics data, as well as the growing qualified evidence of large-scale population-based genomic research. This paper provides a review of genome-wide association research of aging.

Species and characteristics of volatile organic compounds emitted from an auto-repair painting workshop.

Volatile organic compounds (VOCs) are secondary pollutant precursors having adverse impacts on the environment and human health. Although VOC emissions, their sources, and impacts have been investigated, the focus has been on large-scale industrial sources or indoor environments; studies on relatively small-scale enterprises (e.g., auto-repair workshops) are lacking. Here, we performed field VOC measurements for an auto-repair painting facility in Korea and analyzed the characteristics of VOCs emitted from the main painting workshop (top coat). The total VOC concentration was 5069-8058 ppb, and 24-35 species were detected. The VOCs were mainly identified as butyl acetate, toluene, ethylbenzene, and xylene compounds. VOC characteristics differed depending on the paint type. Butyl acetate had the highest concentration in both water- and oil-based paints; however, its concentration and proportion were higher in the former (3256 ppb, 65.5%) than in the latter (2449 ppb, 31.1%). Comparing VOC concentration before and after passing through adsorption systems, concentrations of most VOCs were lower at the outlets than the inlets of the adsorption systems, but were found to be high at the outlets in some workshops. These results provide a theoretical basis for developing effective VOC control systems and managing VOC emissions from auto-repair painting workshops.

[Effects of smoking on the wound healing of stage 4 pressure ulcers in rats].

Objective: To explore the effect of smoking on the wound healing of stage 4 pressure ulcers in rats. Methods: Fifty male Sprague-Dawley rats aged 6-8 weeks were divided into simple pressure ulcer group and smoking+ pressure ulcer group according to the random number table, with 25 rats in each group. After the rats in the smoking+ pressure ulcer group received passive smoking intervention for 12 weeks, an iron plate was placed in the back muscle of each rat in 2 groups, and a magnet was placed outside the skin at the corresponding position of the iron plate for 2 h at each time, with 5 times a day and continuously for 6 days to reproduce stage 4 pressure ulcer model. Immediately after establishing the model, 3 rats in each group were sacrificed and wound tissue was collected, and hematoxylin-eosin staining was applied to observe the pathological changes of the wounds. On 1, 3, 7, and 14 day (s) after establishing the model, 3 rats in each group were collected to measure the pressure ulcer wound area by the paper jam method. After measurement of the wound area, the rats were sacrificed and the wound tissue was collected, and the protein expression levels of matrix metalloproteinases 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in wound tissue were detected by immunohistochemical method, and the ratio of MMP-9/TIMP-1 was calculated.The wound healing time of the remaining 10 rats in each group was recorded. Data were statistically analyzed with analysis of variance for factorial design, two independent sample t test, and Bonferroni correction. Results: (1) Immediately after establishing the model, muscle fiber necrosis and dissolution with large areas were seen on the wound, the myofibrils arranged loosely, and more lymphocytes and monocytes infiltration were seen around the wound of rats in simple pressure ulcer group. A large number of necrotic myofibers were dissolved and gradually disappeared, the myofibrils arranged loosely, and the number of diffuse lymphocytes and monocyte infiltration in wound of rats in smoking+ pressure ulcer group were significantly higher than those in simple pressure ulcer group. (2) The wound areas of rats in smoking+ pressure ulcer group were significantly larger than those in simple pressure ulcer group on 1, 3, 7, and 14 day (s) after establishing the model (t=3.019, 2.549, 2.181, 3.674, P<0.05 or P<0.01). (3) On 1 to 14 days after establishing the model, the protein expression levels of MMP-9 and TIMP-1 in the wound tissue and the ratio of MMP-9/TIMP-1 of rats in the two groups increased first and then decreased. On 1, 3, 7, and 14 day (s) after establishing the model, the protein expression levels of MMP-9 in the wound tissue and the ratio of MMP-9/TIMP-1 of rats in smoking+ pressure ulcer group were significantly higher than those in simple pressure ulcer group (t=4.783, 4.508, 6.325, 7.204, 3.078, 2.989, 4.081, 4.696, P<0.05 or P<0.01), and the protein expression levels of TIMP-1 in wound tissue of rats in the two groups were similar. (4) The wound healing time of rats in smoking+ pressure ulcer group was (48.9±2.6) d, which was significantly longer than (35.2±2.3) d of simple pressure ulcer group (t=12.477, P<0.05). Conclusions: Smoking can up-regulate the expression of MMP-9 in pressure ulcer wound and result in an imbalance of MMP-9/TIMP-1, thereby affecting the wound healing of stage 4 pressure ulcers in rats.

LncRNA AB073614 promotes the proliferation and inhibits apoptosis of cervical cancer cells by repressing RBM5.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "LncRNA AB073614 promotes the proliferation and inhibits apoptosis of cervical cancer cells by repressing RBM5, by L.-Y. Guo, C.-F. Qin, H.-X. Zou, M.-Y. Song, M.-L. Gong, C. Chen, published in Eur Rev Med Pharmacol Sci 2019; 23 (6): 2374-2379-DOI: 10.26355/eurrev_201903_17382-PMID: 30964162" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17382.

Association between C-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma.

OBJECTIVE: To explore the association between c-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma (NHL). PATIENTS AND METHODS: A total of 200 NHL patients in our hospital in the past 3 years were collected as disease group, while 200 healthy people were taken as control group. The genomic deoxyribonucleic acid (DNA) in the peripheral blood was extracted in both groups, amplified via Polymerase Chain Reaction (PCR) and sent to the company for the detection of c-myc and K-ras gene polymorphisms. The expressions of c-myc and K-ras were detected via Reverse Transcription-quantitative PCR (RT-qPCR), and the levels of clinical indexes hemoglobin (Hb), platelet (PLT) and lactate dehydrogenase (LDH) were determined in the Laboratory Department. RESULTS: The allele distribution at c-myc gene locus rs121918684 was different between control group and disease group (p=0.000), and the G allele frequency was 202 (0.505) in the control group and 263 (0.657) in the disease group. In the disease group, the GG genotype frequency at c-myc gene locus rs121918684 [97 (0.485)], the CC genotype frequency at rs775522201 [98 (0.490)], and the GA genotype frequency at K-ras gene locus rs1137188 [127 (0.635)] were all significantly higher than those in the control group (p=0.000, p=0.002, p=0.011). In the disease group, the frequency of recessive model GC+CC (p=0.003), heterozygous model GC (p=0.035), and homozygous model CC (p=0.037) at c-myc gene locus rs121918684 was significantly lower than that in the control group, and the frequency of recessive model CT+TT (p=0.046) at c-myc gene locus rs775522201 was also markedly lower than that in the control group. The haplotype frequency of c-myc CC (p=0.000), GC (p=0.000), and GT (p=0.018) in the disease group was different from that in the control group. Moreover, the CT genotype at c-myc gene locus rs775522201 was remarkably correlated with the c-myc gene expression, and the gene expression was markedly increased in the disease group. The TT genotype at K-ras gene locus rs12245 was correlated with the K-ras gene expression, and the gene expression was notably increased in the disease group. There was an association between GG genotype at c-myc gene locus rs121918684 and LDH level (p=0.000), between CT genotype at c-myc gene locus rs775522201 and PLT level (p=0.002), and between AA genotype at K-ras gene locus rs1137188 and Hb level (p=0.003). CONCLUSIONS: The c-myc and K-ras gene polymorphisms are associated with susceptibility to NHL, gene expression and levels of Hb, PLT, and LDH.

LncRNA AB073614 promotes the proliferation and inhibits apoptosis of cervical cancer cells by repressing RBM5.

OBJECTIVE: Recent studies have determined the crucial role of long noncoding RNAs (lncRNAs) in cancer development. Cervical cancer (CC) is a common type of fatal gynecological cancer worldwide. This study aims to identify the role of lncRNA AB073614 in the progression of CC. PATIENTS AND METHODS: Relative level of AB073614 in 3 CC cell lines and 48 paired CC samples was determined by the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The potential regulatory effects of AB073614 on the cellular behaviors of CC cells were explored through apoptosis assay, colony formation assay, and proliferation assay in vitro. The underlying mechanism of AB073614 in mediating the progression of CC was also conducted. RESULTS: The AB073614 expression was remarkably higher in CC tissues than that in adjacent tissues. The knockdown of AB073614 inhibited proliferation but induced apoptosis of CC cells. In addition, RBM5 (RNA binding motif 5) was upregulated in CC cells after knockdown of AB073614 in vitro. Furthermore, a negative correlation was seen between the expressions of RBM5 and AB073614 in CC tissues. CONCLUSIONS: AB073614 could promote proliferation and inhibit apoptosis of CC cells through downregulating RBM5, which may offer a new therapeutic intervention for CC patients.

[Advances in basic and clinical research on liver cirrhosis in 2016].

Esophageal and gastric varices are common complications of liver cirrhosis and are seen in 50% of patients with liver cirrhosis. The annual incidence rate of esophagogastric variceal bleeding is 5%-15%, and even if the recommended treatment is used, the 6-week mortality rate is still as high as 15%-20%. Spontaneous bacterial peritonitis (SBP) is a common complication of end-stage liver disease and has an incidence rate of 10%-30% in patients with severe liver damage. SBP refers to the bacterial infection of the peritoneum and/or ascites that occurs in the absence of any inflammation in adjacent tissues (e.g., intestinal perforation and intestinal abscess). Hepatic encephalopathy (HE) is the clinical syndrome manifesting as cognitive impairment in patients with chronic liver disease, and its pathogenesis has not yet been fully elucidated and may be associated with ammonia poisoning theory, γ-aminobutyric acid and endogenous benzodiazepine complex receptor theory, and inflammatory pathway theory. This article introduces the advances in the treatment of upper gastrointestinal bleeding in patients with liver cirrhosis, SBP, and HE in 2016.

[Relationship between gestational glucose, lipid metabolism parameters and fetal distress].

OBJECTIVE: To study the relationship between gestational glucose, lipid metabolism parameters and fetal distress. METHODS: Retrospectively, 82 cases of primipara with fetal distress and 246 cases of primipara without any obstetric complications were analyzed. The latter were treated as control group. All the patients were from the same hospital between January, 2014 and January, 2016. Factors as fasting plasma-glucose (FPG), triglyceride (TG), total cholesterol (TC),high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C) parameters during early pregnancy, blood glucose of fasting, 1 hour and 2 hours in 75 g oral glucose tolerance test (OGTT), at 24 to 28 weeks of gestation, hemoglobin and blood pressure parameters during ante partum were recorded for all the cases while parameters as glucose and lipid, hemoglobin, blood pressure and cesarean section rate were compared between the 2 groups. RESULTS: FPG during early gestation, blood glucose of fasting, 1 hour and 2 hours 75 g OGTT during mid gestation, systolic blood pressure and diastolic blood pressure during late gestation in the'fetal distress'group were significantly higher than that of the control group. Hemoglobin during late gestation in the fetal distress group was lower than that of the control group, with statistically significant difference (P<0.05). The rate of cesarean section in the fetal distress group was higher than that in the control group (χ(2)=4.489, P=0.034). CONCLUSIONS: High BMI at pre-pregnancy, high blood glucose during early and mid-gestation, high blood pressure and anemia during late gestation were related to the fetal distress group. Cesarean section rate was high for pregnancy women with fetal distress.

Effect of cocaine on ion channels and glutamatergic EPSCs in noradrenergic locus coeruleus neurons.

The locus coeruleus (LC) is an important brainstem area involved in cocaine addiction. However, evidence to elucidate how cocaine modulates the activity of LC neurons remains incomplete. Here, we performed whole recordings in brain slices to evaluate the effects of cocaine on the sodium (Na(+)), potassium (K(+)), calcium (Ca(2+)) channels, and glutamatergic synaptic transmission in the locus coeruleus neurons. Local application of cocaine significantly and reversibly reduced the spontaneous firing rate but did not affect action potential amplitude, rising time, decay time, or half width of noradrenergic locus coeruleus neurons. Moreover, cocaine attenuated the sodium current but did not affect potassium and calcium currents. The N-methyl-D-aspartate receptor mediated excitatory postsynaptic currents were reduced by neuropeptide galanin but not cocaine. All those data demonstrate that cocaine has inhibitory effect on the spontaneous activities and sodium current in locus coeruleus neurons. Therefore, neuromodulation of sodium channel in locus coeruleus neurons may play an important role in drug addiction.

Branched oligomerization of cell-permeable peptides markedly enhances the transduction efficiency of adenovirus into mesenchymal stem cells.

Cell-permeable peptides (CPPs) promote the transduction of nonpermissive cells by recombinant adenovirus (rAd) to improve the therapeutic efficacy of rAd. In this study, branched oligomerization of CPPs significantly enhanced the transduction of human mesenchymal stem cells (MSCs) by rAd in a CPP type-independent manner. In particular, tetrameric CPPs increased transduction efficiency at 3000-5000-fold lower concentrations than did monomeric CPPs. Although branched oligomerization of CPPs also increases cytotoxicity, optimal concentrations of tetrameric CPPs required for maximum transduction are at least 300-1000-fold lower than those causing 50% cytotoxicity. Furthermore, although only approximately 60% of MSCs were maximally transduced at 500 muM of monomeric CPPs, >95% of MSCs were transduced with 0.1 muM of tetrameric CPPs. Tetrameric CPPs also significantly increased the formation and net surface charge of CPP/rAd complexes, as well as the binding of rAd to cell membranes at a greater degree than did monomeric CPPs, followed by rapid internalization into MSCs. In a critical-size calvarial defect model, the inclusion of tetrameric CPPs in ex vivo transduction of rAd expressing bone morphogenetic protein 2 into MSCs promoted highly mineralized bone formation. In addition, MSCs that were transduced with rAd expressing brain-derived neurotrophic factor in the presence of tetrameric CPPs improved functional recovery in a spinal cord injury model. These results demonstrated the potential for tetrameric CPPs to provide an innovative tool for MSC-based gene therapy and for in vitro gene delivery to MSCs.