[A meta-analysis of risk factors for multidrug-resistant tuberculosis in China].

Objective: To explore the main risk factors of multidrug-resistant tuberculosis (MDR-TB) in China and to provide evidence-based evidence for MDR-TB preventon and control. Methods: All relevant literatures were searched in thedatabases, such as Pubmed, Web of Science and CNKI, Wanfang, VIP and SinoMed from 2000 to 2021. Quality evaluation and data extraction were carried out, and then a meta-analysis was performed using Stata 16.0 software. Results: A total of 59 literatures (36 cross-sectional and 23 case-control) including 75 793 participants were included in this study, and meta-analysis results showed age (OR=1.27, 95%CI: 1.05-1.54), education level (OR=1.29, 95%CI: 1.02-1.65), positive sputum smear (OR=2.56, 95%CI: 1.09-6.04), pulmonary cavity (OR=1.99, 95%CI: 1.57-2.52), course of disease (OR=4.25, 95%CI: 1.95-9.30), history of tuberculosis treatment (OR=6.42,95%CI:5.40-7.63), treatment interruption (OR=2.81, 95%CI: 1.50-5.29), irregular medication (OR=5.02, 95%CI: 2.95-8.54), adverse drug reactions (OR=4.27, 95%CI: 2.22-8.19), combined chronic obstructive pulmonary disease (COPD) (OR=2.21, 95%CI: 1.45-3.37), tuberculosis exposure history (OR=1.99, 95%CI: 1.36-2.91), smoking history (OR=1.35, 95%CI: 1.09-1.66) and floating population (OR=1.60, 95%CI: 1.04-2.44) were associated with the occurrence of MDR-TB. Conclusions: The high risk groups were farmer, low education level, pulmonary cavity, long course of disease, history of tuberculosis treatment, treatment interruption, irregular medication, adverse drug reaction, co-COPD, contact history of tuberculosis, smoking history, rural residence, and floating population. We should pay attention to high-risk groups, strengthen management and take effective measures such as early screening, knowledge education on tuberculosis, standardized and personalized treatment and whole-course supervision.

Sildenafil might impair maternal-fetal immunotolerance by suppressing myeloid-derived suppressor cells in mice.

Myeloid-derived suppressor cells (MDSCs) as an important population of immune cells were found to restrain T cell function, polarize T-helper cells (Th) 1/Th2 toward Th2 response and induce regulatory T cells (Tregs), therefore enhancing the immunotolerance during pregnancy. Sildenafil has been applied for poor endometrial quality in implantation failure patients. Nevertheless, investigations have shown that sildenafil could reduce MDSCs-dependent immunosuppression. Whether sildenafil affects embryo implantation by suppressing MDSCs? To address this question, using the mice model, we investigated the amounts of immune cells in peripheral blood and endometrial cells from control group (CG), sildenafil low-dose group (LDG) and high-dose group (HDG). We found that both treatment groups displayed a marked deficiency in polymorphonuclear (PMN)-MDSCs and Th2 from mice blood and endometrium as compared to these from CG. The frequency of Tregs in endometrium from HDG was lower than those from CG. Th1/Th2 ratio in both periphery and uterus from study groups showed a significant increase as compared to those from CG. By relevance analysis, we found that the level of Tregs positively correlated with the level of PMN-MDSCs, whereas the Th1/Th2 ratio negatively correlated with the frequency of PMN-MDSCs in uterus. Moreover, there was a positive relationship between the amount of blood PMN-MDSCs and endometrial PMN-MDSCs. These results suggest that we should carefully weigh the pros and cons of using sildenafil when applied to patients with poor endometrial receptivity.

Long non-coding RNA LINP1 functions as an oncogene in endometrial cancer progression by regulating the PI3K/AKT signaling pathway.

OBJECTIVE: Endometrial cancer (EC) accounts for about 6% of new cancer cases in female and about 3% of cancer-related deaths were caused by EC. The poor prognosis is mainly due to the distant spread and poor differentiation. In the current study, we want to figure out the role of long non-coding RNA (LncRNA) LINP1 in EC progression. PATIENTS AND METHODS: The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay was involved to access the expression level of LINP1 in EC cell lines and tissues. The Cell Counting Kit-8 (CCK-8) assay, colony formation assay, transwell and Matrigel assay were recruited to figure out the ability of LINP1 in cell proliferation and metastasis in EC. Subsequently, Western blotting was used to detect the expression level of PI3K/AKT in EC. Besides, we used the tumor formation assay in vivo to examine the ability of LINP1 in tumor formation in vivo. RESULTS: LINP1 was proved to be up-regulated in EC cell lines and tissues by qRT-PCR assay. CCK-8 assay and colony formation assay were conducted and the results indicated that LINP1 over-expression can promote cell proliferation in EC in vitro. The data of transwell and Matrigel assays indicated that up-regulated LINP1 can facilitate cell migration and invasion. The results of Western blotting validated that LINP1 can activate PI3K/AKT signaling. Besides, the tumor formation assay verified that LINP1 can promote tumor formation in vivo. CONCLUSIONS: Our research validated that LINP1 served as an oncogenic role in EC progression. The PI3K/AKT signaling pathway might be the underlying mechanism of EC progression. We hope our study can provide novel treatment targets and biomarkers in EC development and progression.

[Incipient observation of cornea astigmatism after combined cataract phacoemulsification and Callisto Eye-assisted arcuate keratotomy].

Objective: To research the change of cornea astigmatism after combined cataract phacoemulsification and Callisto Eye-assisted arcuate keratotomy and to assess the effectiveness and safety of this surgery for improving patients' uncorrected visual acuity. Methods: Prospective interventional non-randomized controlled study. Consecutive cataract patients with cornea astigmatism greater than 0.75 D were recruited at the southern part of Beijing Tongren Hospital, Capital Medical University and Beijing Tongren Eye Center, between October 2017 and March 2018, and were divided into the experimental group and the controlled group. In the experimental group the patients are further divided into three subgroups of A(0.75-1.24 D), B(1.25-1.74 D) and C(≥1.75 D) according to the degrees of their cornea astigmatism before surgery. The combined cataract phacoemulsification and Callisto Eye-assisted arcuate keratotomy were performed for the experimental group with the depth and length of the corneal arcuate incision differing in subgroups, while only cataract phacoemulsification was performed for the control group. The complications on and after the surgery were observed and the change of cornea astigmatism and uncorrected visual acuity in three months after surgery were recorded. Independent sample t test was used for statistical analysis. Results: A total of 108 patients (108 eyes) were enrolled, including 46 males and 62 females, the age was (68±11) years old.The experimental group included 76 eyes of 76 patients (subgroup A 38 eyes, subgroup B 18 eyes, subgroup C 20 eyes). The controlled group included 32 eyes of 32 patients. The preoperative cornea astigmatism and postoperative cornea astigmatism of the experimental group are (1.54±0.90) D and 0.62 (0.36-0.95) D. The change of cornea astigmatism of the experimental group is (1.15±0.58) D. The index of correction and flattening are 0.97±0.20 and 0.61±0.31 respectively, the error of axial deviation is 9.55°±5.22°. The index of success is 0.51±0.36. In the experimental group, subgroup C has the highest correction efficiency of corneal astigmatism (axial deviation error 8.07°±3.13°, flattening index 0.72±0.31) which is followed by subgroup B, and subgroup A ranked the last one (axial deviation error 10.27°±6.47°, flattening index 0.42±0.30).The uncorrected distance visual acuity (logarithm of the minimum angle of resolution) of the experimental group and the control group are significantly different (0.15±0.12 vs. 0.24±0.17, t=-4.129, P<0.01). None of the patients recruited have complications including poor wound healing and cornea penetration and cornea edema and cystoid macular edema on or after the surgery. Conclusions: Combined cataract phacoemulsification and Callisto Eye-assisted arcuate keratotomy are of favorable effect on improving the uncorrected visual acuity and have relatively good accuracy and predictability on the correction of cornea astigmatism. It is a safe surgery with considerable efficency. (Chin J Ophthalmol, 2019, 55: 502-508).

Clinicopathological and prognostic significance of miR-4317 expression in gastric cancer patients.

OBJECTIVE: Previous studies have revealed that miR-4317 was abnormally expressed and functioned as a tumor suppressor in several tumors, including gastric cancer (GC). However, the clinical significance of miR-4317 in GC remains largely unclear. Our present study aimed at investigating the possibility of miR-4317 as a novel prognostic biomarker for GC patients. PATIENTS AND METHODS: RT-PCR was performed to determine the levels of miR-4317 in GC tissues and matched normal gastric tissues. Association between miR-4317 levels and clinicopathological factors was also analyzed using Chi-square test. Kaplan-Meier survival analysis and univariate and multivariate assays were further conducted to investigate the correlation between miR-4317 expression and GC patients prognosis. RESULTS: We showed that miR-4317 expression levels were significantly lower in GC cell lines compared to matched normal tissues (p< 0.01). Down-regulation of miR-4317 was observed to be associated with lymph node metastasis (p = 0.019), distant metastasis (p = 0.011) and TNM stage (p = 0.007). In addition, Kaplan-Meier analysis showed that patients with low miR-4317 expression had a shorter overall survival compared with the high miR-4317 expression group (p = 0.0009). Furthermore, multivariate analysis revealed that miR-4317 expression was an independent prognostic marker of overall survival of GC patients. CONCLUSIONS: miR-4317 expression may be a novel and valuable prognostic factor in GC.

Clinical and genetic characteristics of familial hypercholesterolemia patients with type 2 diabetes.

PURPOSE: Though type 2 diabetes mellitus (T2DM) is an important and independent risk factor for coronary artery disease (CAD) in the general population, the impact of T2DM on CAD in patients with familial hypercholesterolemia (FH) is less understood. Thus, the current study aimed to examine the features of FH patients with T2DM and explore the effects of T2DM on CAD in FH. METHODS: A total of 289 clinical heterozygous FH (HeFH) patients diagnosed with Dutch Lipid Clinic Criteria were consecutively recruited and divided into a T2DM group (n = 58) and non-T2DM group (n = 231). Clinical characteristics and laboratory findings were compared between the two groups. Target exome sequencing was used for gene mutation analysis. RESULTS: HeFH patients with T2DM had significantly higher levels of triglycerides, body mass index and free fatty acids than did non-T2DM patients; moreover, patients with T2DM more frequently exhibited hypertension. However, the spectrum of FH-causing mutations was not significantly different (p = 0.061). Notably, patients with T2DM had higher prevalence of CAD (p = 0.012) and higher Gensini Score (p = 0.002). The regression analysis confirmed that HbA1c was an independent risk factor for both the presence and severity of CAD [OR 2.321 (1.098-4.904), p = 0.027; OR 1.349 (1.032-1.762), p = 0.028, respectively] in patients with HeFH. CONCLUSIONS: Although there were not many differences in the clinical, lipid and genetic aspects of HeFH patients with and without T2DM, T2DM and HbA1c were associated with worse coronary lesions, suggesting that diabetes and the degree of blood glucose control are also important determinants of cardiovascular disease in these patients.

[Value the correction of corneal astigmatism in cataract surgery].

The aim of modern micro-incision phacoemulsification combined with foldable intraocular lens implantation and femtosecond laser-assisted cataract surgery is evolving from a simple pursuit of recuperation to a refractive procedure, which involves the correction of ametropia according to preoperative and postoperative refractive conditions, especially corneal astigmatism, in order to achieve the goal of optimized postoperative uncorrected full range of vision. Nowadays, due attention to the effect of preoperative corneal astigmatism, surgery-induced astigmatism and residual astigmatism after operation is lacked, which affect postoperative visual acuity significantly. There are many effective ways to reduce corneal astigmatism after cataract surgery including selecting appropriate size and location of clear corneal incision, employing astigmatism keratotomy and the implantation of Toric intraocular lenses, which need to be appropriately applied and popularized. At the same time, surgical indications, predictability and safety should also be taken into account.(Chin J Ophthalmol, 2018, 54: 321-323).

miR-424 protects PC-12 cells from OGD-induced injury by negatively regulating MKP-1.

OBJECTIVE: It's of great significance to investigate the novel targets of drugs for the treatment of stroke. In this study, we explored the neuroprotective role of miR-424 in oxygen glucose deprivation (OGD)-induced injuries in PC-12 cells. MATERIALS AND METHODS: PC-12 cells were subjected to OGD stimulation to mimic ischemic injury. The expressions of miR-424 and mitogen-activated protein kinase phosphatase-1 (MKP-1) were altered by transient transfection with miR-424 mimic, miR-424 inhibitor, pEX-MKP-1, or sh-MKP-1. Cell counting kit-8 (CCK-8) assay, flow cytometry, and quantitative reverse transcription polymerase chain reaction (qRT-PCR), were conducted to respectively detect cell viability, apoptotic cells, and the expression of miR-424 and MKP-1. The protein expressions of several factors were determined by Western blot. Meanwhile, relative luciferase activity assay was done to verify the predicted targets association. RESULTS: OGD induced injury in PC-12 cells by suppressing cell viability and inducing apoptosis. OGD also induced the expression of miR-424 in PC-12 cells. Overexpression of miR-424 protected PC-12 cells from OGD-induced injury by increasing cell viability and decreasing apoptosis. MKP-1 was a direct target of miR-424, and its expression was negatively regulated by miR-424. Up-regulation of expression of MKP-1 aggravated OGD-induced cell injury by inhibiting the expression of hypoxia-inducible factor 1α (HIF-1α), thus inhibiting the PI3K/AKT/mTOR pathways. CONCLUSIONS: miR-424 protected PC-12 cells from OGD-induced injury through direct suppression of MKP-1 expression, as MKP-1 promoted OGD-induced cell injury by inhibiting the expression of HIF-1α and PI3K/AKT/mTOR pathways.

Abdominal wall endometriosis occurring after cesarean section: an underestimated complication.

The aim of the study was to review patients characteristics, describe the exact anatomic locations and size of the endometriosis in the abdominal wall, and discuss the factors that may contribute to mesh use during abdominal wall endometriosis (AWE) resection. MATERIALS AND METHODS: Patients diagnosed with AWE in their surgical scars from January 2008 to December 2014 were documented. Descriptive data was collected and analyzed. RESULTS: A total of 95 patients with an age ranging from 26 to 48 years, with a mean age of 33.5 ?5.0 years at the time of excision were analyzed. The mean diameter of the mass was 3.25 cm in the present series with an average of 4.97 cm in the mesh group by ultrasound. A total of 18 patients had mesh therapy for fascia defect compared with 77 non-mesh therapy patients. The size of the lesions, the mean duration of symptoms for painful mass, and level of the serum CA125 were statistically different between mesh group and non-mesh group (p < 0.05). Cases of endometriosis lesions limited to the adipose layer had significant lower chance of using mesh (p < 0.05). However, adipose layer endometriosis lesions that had penetrated through the fascia layer and invaded into rectus abdominis muscle layer with/without peritoneum layer had significant higher chance of using mesh (p < 0.05). CONCLUSIONS: The more common position for scar endometriosis may be in the adipose layer at the corner of the surgical scar. Mesh therapy should be considered before surgery when the diameter of the abdominal wall mass detected by ultrasound is more than five em and/or when the lesions invade into rectus abdominis muscle with/without peritoneum tissues from adipose and fascia layers.

Downregulation of miR-320a/383-sponge-like long non-coding RNA NLC1-C (narcolepsy candidate-region 1 genes) is associated with male infertility and promotes testicular embryonal carcinoma cell proliferation.

Long non-coding RNAs (lncRNAs), which are extensively transcribed from the genome, have been proposed to be key regulators of diverse biological processes. However, little is known about the role of lncRNAs in regulating spermatogenesis in human males. Here, using microarray technology, we show altered expression of lncRNAs in the testes of infertile men with maturation arrest (MA) or hypospermatogenesis (Hypo), with 757 and 2370 differentially down-regulated and 475 and 163 up-regulated lncRNAs in MA and Hypo, respectively. These findings were confirmed by quantitative real-time PCR (qRT-PCR) assays on select lncRNAs, including HOTTIP, imsrna320, imsrna292 and NLC1-C (narcolepsy candidate-region 1 genes). Interestingly, NLC1-C, also known as long intergenic non-protein-coding RNA162 (LINC00162), was down-regulated in the cytoplasm and accumulated in the nucleus of spermatogonia and primary spermatocytes in the testes of infertile men with mixed patterns of MA compared with normal control. The accumulation of NLC1-C in the nucleus repressed miR-320a and miR-383 transcript and promoted testicular embryonal carcinoma cell proliferation by binding to Nucleolin. Here, we define a novel mechanism by which lncRNAs modulate miRNA expression at the transcriptional level by binding to RNA-binding proteins to regulate human spermatogenesis.

Hydrogen sulphide induces vasoconstriction of rat coronary artery via activation of Ca(2+) influx.

AIM: Hydrogen sulphide (H2S) exhibits a dual modulation of isolated artery tension. This study investigated the vasoconstrictive effect of sulphur sodium hydride (NaHS), a donor of gaseous H2S, on rat coronary artery. METHODS: The contractile response of isolated arteries was recorded using a wire myograph. Fluo-3/AM was used to load vascular smooth muscle, and intracellular calcium was determined using confocal laser microscopy. The protein expression of Rho kinase was examined using Western blot. RESULTS: NaHS induced concentration-dependent contractions of rat coronary artery, and the contraction reached approx. 65% of 60 mm KCl-induced contraction. The NaHS-induced contraction was elevated following the removal of endothelium or the use of the nitric oxide synthase inhibitor L-NAME. The cyclooxygenase inhibitor indomethacin reduced NaHS-induced contraction. The Rho kinase inhibitor Y-27632 significantly attenuated NaHS-induced vasoconstriction. Furthermore, NaHS elevated the protein expression of Rho kinase. NaHS-induced contraction was completely abolished in a Ca(2+)-free solution and suppressed by the Ca(2+) influx blocker nifedipine (100 nm). NaHS also significantly increased the change rate of Ca(2+) fluorescence intensity. However, treatment with a Cl(-)/HCO(3-) exchanger blocker, K(+) channel blockers, the mitogen-activated protein kinase inhibitor U-0126 or cyclic adenosine monophosphate did not affect contraction. Species-dependent differences in NaHS-induced vasoconstriction were observed because these effects were only modest in dog coronary artery and absent in rabbit coronary artery. CONCLUSIONS: NaHS induces the contraction of rat coronary artery, which is dependent on the activation of Ca(2+) influx. Rho kinase likely participates in the vasoconstriction.

The (TTTA)n polymorphism in intron 4 of CYP19 and the polycystic ovary syndrome risk in a Chinese population.

Polycystic ovary syndrome (PCOS) is a heterogeneous disease with a strong genetic origin, but the specific determinants are still unknown. The aim of this study was to investigate the association between the (TTTA) n polymorphism in intron 4 of CYP19 and the PCOS risk in a Chinese population. We performed a case-control study which involved 222 PCOS patients and 281 controls. The fluorescent-labeled target DNA fragments containing the (TTTA)n short tandem repeats were obtained by PCR, thereafter genotyped via capillary electrophoresis. Representative alleles were directly sequenced to confirm their repeat numbers. Genotype analysis revealed seven different alleles including 7-3(∆)-, 7-, 8-, 10-, 11-, 12- and 13-TTTA-repeats. The most common allele in a Chinese population is (TTTA) 11 in our study (0.354 for PCOS and 0.390 for controls). PCOS patients showed a higher frequency of short alleles compared with controls (0.47 vs. 0.41, OR=1.245, 95% CI 0.97-1.60). The overall allelic distributions of this polymorphism did not show any significant differences between PCOS patients and the control group. No statistical differences were found in the clinical parameters or serum steroid hormone levels among PCOS patients with different genotypes. In conclusion, PCOS patients had a higher frequency of short alleles, albeit this might not strongly affect the risk of PCOS.

The targeting and functions of miRNA-383 are mediated by FMRP during spermatogenesis.

Our previous studies have shown that microRNA-383 (miR-383) expression is downregulated in the testes of infertile men with maturation arrest (MA). Abnormal testicular miR-383 expression may potentiate the connections between male infertility and testicular germ cell tumors. However, the mechanisms underlying the targeting and functions of miR-383 during spermatogenesis remain unknown. In this study, we found that fragile X mental retardation protein (FMRP) was associated with 88 miRNAs in mouse testis including miR-383. Knockdown of FMRP in NTERA-2 (NT2) (testicular embryonal carcinoma) cells enhanced miR-383-induced suppression of cell proliferation by decreasing the interaction between FMRP and miR-383, and then affecting miR-383 binding to the 3'-untranslated region of its target genes, including interferon regulatory factor-1 (IRF1) and Cyclin D1 both in vivo and in vitro. On the other hand, FMRP levels were also downregulated by overexpression of miR-383 in NT2 cells and GC1 (spermatogonia germ cell line). miR-383 targeted to Cyclin D1 directly, and then inhibited its downstream effectors, including phosphorylated pRb and E2F1, which ultimately resulted in decreased FMRP expression. Reduced miR-383 expression, dysregulated cyclin-dependent kinase 4 expression (one of the downstream genes of miR-383) and increased DNA damage were also observed in the testes of Fmr1 knockout mice and of MA patients with a downregulation of FMRP. A potential feedback loop between FMRP and miR-383 during spermatogenesis is proposed, and FMRP acts as a negative regulator of miR-383 functions. Our data also indicate that dysregulation of the FMRP-miR-383 pathway may partially contribute to human spermatogenic failure with MA.

Secondhand cigarette smoke exposure causes upregulation of cerebrovascular 5-HT(1) (B) receptors via the Raf/ERK/MAPK pathway in rats.

AIM: Cigarette smoke exposure increases the risk of stroke. Upregulation of 5-hydroxytryptamine 1B (5-HT(1) (B) ) receptors is associated with the pathogenesis of cerebral ischaemia. This study examined the hypothesis that the expression of 5-HT(1) (B) receptors is altered in brain vessels after secondhand smoke (SHS) exposure. METHODS: Rats were exposed to SHS in vivo for 200 min daily for 8 weeks. The contractile responses of isolated cerebral arteries were studies by a sensitive myograph. The mRNA and protein expression for 5-HT(1) (B) receptors were examined by real-time PCR, Western blot and immunofluorescence respectively. In addition, the phosphorylation of Raf/extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinases (MAPK) pathway was evaluated. RESULTS: The results showed that SHS exposure shifted the 5-HT(1) (B) receptor-mediated concentration-contraction curve towards the left with a markedly increased maximal contraction. Furthermore, there were significant elevations in mRNA level and protein expression of 5-HT(1) (B) receptors in SHS-exposed rats. Immunostaining revealed that the 5-HT(1) (B) receptors were localized to the smooth muscle cells of cerebral arteries. SHS was also found to induce the phosphorylation of Raf-1 and ERK1/2 proteins. The administration of a Raf-1 inhibitor GW5074 attenuated the 5-HT(1) (B) receptor upregulation. CONCLUSION: Secondhand smoke exposure upregulates cerebrovascular 5-HT(1) (B) receptors in rats. The receptor upregulation is associated with Raf/ERK/MAPK activation.

Improvement of heart allograft acceptability associated with recruitment of CD4+CD25+ T cells in peripheral blood by recipient treatment with granulocyte colony-stimulating factor.

BACKGROUND: Granulocyte colony-stimulating factor (G-CSF), an important hematopoietic growth factor of the myeloid lineage, exerts profound immunoregulatory effects in T-cell tolerance. The study objective was to investigate the potential mechanism of G-CSF's antirejection effects in a fully mismatched rat cardiac allograft model. METHODS: The allograft recipients were treated with subcutaneous injection of recombinant human G-CSF (rh-G-CSF) at a dose of 250 microg/kg/d for 6 days starting from the day of cardiac transplantation. The alloreactive T-cell response and rejection level of G-CSF-treated rats were compared with those of control rats using mixed lymphocyte reactions (MLR) and histological examinations. Cytokine and cellular profiles were determined using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The presence and suppressive functions of regulatory T cells were determined by adoptive cell transfer experiments. RESULTS: Posttransplantation treatment of recipients with rh-G-CSF alone prolonged allograft survival, improved allograft biopsy grading scores, and induced alloreactive T-cell hyporesponsiveness accompanied by high levels of interleukin-10 (IL-10) and transforming growth factor-beta1 (TGF-beta1) production in MLR. It also enhanced CD4+CD25+ T cells in peripheral blood. The splenocytes from rh-G-CSF-treated recipients transferred antirejection effects to secondary recipients. CONCLUSIONS: Posttransplantation treatment of cardiac allograft recipients with rh-G-CSF leads to alloreactive T-cell hyporesponsiveness in vivo and in vitro associated with recruitment of CD4+CD25+ T cells in the peripheral blood. This study may provide insight into the application of G-CSF to control acute rejection of solid organ transplantations.

Expression of TRAIL, DR4, and DR5 in kidney and serum from patients receiving renal transplantation.

Renal transplantation is the best treatment of some end-stage renal diseases. Unfortunately, not every transplant is successful due to the rejection or dysfunction of the transplanted kidney. Many cytokines participate in rejection by inducing inflammation or apoptosis. In this study, the expressions of TRAIL, DR4, and DR5 in rejected renal tissue and of serum soluble TRAIL (sTRAIL) in patients with kidney rejection were investigated by immunohistochemical staining and sandwich enzyme-linked immunosorbent assay, respectively. The results showed that the expression of TRAIL, DR4 and DR5, and serum sTRAIL levels were markedly upregulated among renal transplant patients. Since both membrane and soluble forms of TRAIL can induce apoptosis of DR4/DR5-expressing cells via recruiting FADD and caspase 8, elevated TRAIL and its receptors may participate in renal graft rejection.

Cytosine methylation of an Sp1 site contributes to organ-specific and cell-specific regulation of expression of the lung epithelial gene t1alpha.

Several recent observations have suggested that cytosine methylation has a role in the in vivo transcriptional regulation of cell-specific genes in normal cells. We hypothesized that methylation regulates T1alpha, a gene expressed primarily in lung in adult rodents. In fetuses T1alpha is expressed in several organs, including the entire nervous system, but during development its expression is progressively restricted to lung alveolar type I epithelial cells, some osteoblasts and choroid plexus. Here we report that T1alpha is methylated at a key Sp1 site in the proximal promoter in cells and organs, including brain, where no gene expression is detectable. Conversely, in T1alpha-expressing cells, these sites are not methylated. In embryonic brain T1alpha is unmethylated and expressed; in adult brain the gene is methylated and not expressed. In lung epithelial cell lines, methylation of the T1alpha promoter in vitro decreases expression by approx. 50% (the maximum suppression being 100%). Analysis of mutated promoter constructs indicates that a single Sp1 site in the proximal promoter provides all or most of the methylation-sensitive gene silencing. We conclude that, in addition to regulation by transcription factors, cytosine methylation has a role in the complex expression patterns of this gene in intact animals and primary cells.

1.3 kilobases of the lung type I cell T1alpha gene promoter mimics endogenous gene expression patterns during development but lacks sequences to enhance expression in perinatal and adult lung.

The T1alpha gene is one of few markers for the type I cell phenotype in the adult mammalian lung. Type I cells form a large, thin epithelial layer that facilitates gas exchange and transport of fluids between the air spaces and capillaries. The T1alpha gene has a complex pattern of developmental expression in lung and brain; in vitro studies indicate that expression is regulated in part by thyroid transcription factor 1, forkhead proteins, and Sp1/Sp3 proteins. To explore the mechanisms that confine T1alpha expression in intact adult animals to alveolar type I and choroid plexus epithelial cells, we generated mice bearing a 1.3-kb T1alpha promoter-chloramphenicol acetyltransferase (CAT) gene. In situ hybridization and RNase protection assays show that the 1.3-kb promoter confers a pattern of CAT expression that largely matches the endogenous T1alpha in embryos and mid-term fetuses in lung and central nervous system. However, the 1.3-kb promoter lacks elements important for perinatal up-regulation of T1alpha in the lung and maintenance of that expression in the adult lung and brain. The final adult pattern of T1alpha expression may be directed by elements outside the 1.3-kb fragment, perhaps those 5' to the 1.3-kb fragment as we show herein, or in 3' and intronic regions. Dev Dyn 1999;215:319-331.