Andrographolide and its derivatives are effective compounds for gastrointestinal protection: a review.

OBJECTIVE: Andrographolide and its derivatives have many functions, such as anti-infection, anti-tumor, neuroprotection, and immune regulation. However, the gastrointestinal protective effects, especially gastrointestinal tumors, and inflammation-related diseases of andrographolide and its derivatives have not been well summarized and discussed. In this review, we aimed to summarize and discuss the pharmacological effects and underlying mechanisms of andrographolide and its derivatives in gastrointestinal protection, with a view to revealing more possibilities of andrographolide and its derivatives in gastrointestinal diseases prevention therapy. MATERIALS AND METHODS: The data in this review are searched and selected from PubMed with the keywords: Andrographolide and Andrographolide derivatives, and relevant data with gastrointestinal protection are extracted and discussed. RESULTS: Andrographolide and its derivatives have prophylactic and therapeutic effects in gastrointestinal disorders such as GU, gastric cancer, colorectal cancer, and inflammatory bowel disease. CONCLUSIONS: Andrographolide and its derivatives are effective compounds for gastrointestinal protection.

Long non­coding RNA XIST promotes cell proliferation of pancreatic cancer through miR­137 and Notch1 pathway.

OBJECTIVE: Long non-coding ribonucleic acids X-inactive specific transcript (lncRNA XIST) is one lncRNAs which involved in multiple human cancers. However, the functions and potential molecular regulatory mechanisms of XIST/microRNA-137 (miR­137) in pancreatic cancer (PC) still need to explore. PATIENTS AND METHODS: PC tissues and cell lines were analyzed for XIST, miR-137 and Notch1 expressions through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Nude mouse xenograft tumor assay was used to detect XIST effects on pancreatic tumorigenesis in vivo. Cell Counting Kit (CCK-8) assay was performed to detect PC cell proliferation. Dual-Luciferase reporter assay, qRT-PCR, RNA immunoprecipitation (RIP) and Western blot assays were applied to validate the target relationship of XIST, miR­137 and Notch1. RESULTS: Results demonstrated that XIST expression was increased in PC tissues and cells. XIST knockdown inhibited PC cell proliferation in vitro and also repressed the tumor growth in vivo. XIST directly interacted with miR-137 and negatively regulated its expression. Notch1 was identified as a target gene of miR-137 and XIST acted as a competitive endogenous RNA (ceRNA) to positively regulate Notch1 expression by suppressing miR-137. In addition, we detected miR-137 was negatively correlated with XIST and Notch1 respectively, and a positive correlation between Notch1 and XIST expression in PC tissues. Furthermore, Notch1 overexpression could offset the suppressing effect of XIST knockdown or miR-137 overexpression on cell proliferation. Therefore, XIST may play an important role in promoting cell proliferation through miR­137 and Notch1 pathway in PC. CONCLUSIONS: To sum up, these results proposed that XIST functioned as an endogenous sponge in promoting PC cell proliferation through competing for miR-137 to regulate Notch1 expression, and may provide more therapeutic targets for the patients with PC.

MiR-16 inhibits pituitary adenoma cell proliferation via the suppression of ERK/MAPK signal pathway.

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, "MiR-16 inhibits pituitary adenoma cell proliferation via the suppression of ERK/MAPK signal pathway, by D.-W. Wang, Y.-Q. Wang, H.-S. Shu, published in Eur Rev Med Pharmacol Sci 2018; 22 (5): 1241-1248-DOI: 10.26355/eurrev_201803_14464-PMID: 29565480" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/14464.

[Single-center study of the abnormal concentration of bone imaging agent and prostate cancer bone metastasis prediction model].

Objective: To investigate the relationship between total prostate specific antigen (TPSA), free prostate specific antigen/total prostate specific antigen [RAT (F/T)], Gleason score, other factors and the whole-body bone plane imaging which was used to evaluate the bone metastasis of prostate cancer (PCa), and the diagnostic value of the abnormal concentration of bone imaging agent for single lesion. Methods: A retrospective analysis of (99)Tc(m)-methylene diphosphonate ((99)Tc(m)-MDP) whole-body bone imaging data of 93 patients with confirmed PCa in The First Hospital of Shanxi Medical University from Jan 2018 to Jan 2019 was conducted. The bone metastasis was diagnosed by whole-body bone imaging. The factors related to PCa bone metastasis, including age, TPSA, RAT (F/T), Gleason score were analyzed by Chi-square test and logistic two-class regression. The optimal cut-off point of TPSA was defined by receiver operating characteristic (ROC) curve. The region of interest (ROI) technique was used to repeatedly delineate the lesion (T) and the background area (NT) outside the bone and calculate the abnormal concentration value of bone imaging agent (T-NT)/NT, and the ROC curve was used to determine its diagnostic value. Results: The result of Chi-square analysis showed that Gleason score, TPSA and RAT (F/T) were associated with bone metastasis (P<0.05). Logistic regression analysis showed that TPSA and RAT (F/T) were associated with bone metastasis (P<0.01). TPSA >92.82 ng/ml was the best diagnosis for bone metastasis, and the sensitivity and specificity were 77.1% and 81.0%, respectively. There were 320 sites of high concentration of imaging agents in the whole-body bone imaging of PCa patients (194 in the metastatic group and 126 in the non-metastasis group). The (T-NT)/NT in the bone metastasis group was 7.11±0.29, the non-bone metastasis group was 2.69±0.20. (T-NT)/NT >3.52 was the best diagnosis for bone metastasis of single lesion, and the sensitivity and specificity were 86.1% and 80.2%, respectively. Conclusions: Gleason score, RAT (F/T) and TPSA are important risk factors of PCa bone metastasis. TPSA >92.82 ng/ml is the most supportive diagnosis for PCa bone metastasis. The abnormal concentration of bone imaging agent >3.52 owns the best diagnosis effect for the single lesion of PCa.

Knockdown of long non-coding RNA LINC00518 inhibits cervical cancer proliferation and metastasis by modulating JAK/STAT3 signaling.

OBJECTIVE: Long intergenic non-protein coding RNA 518 (LINC00518) was reported to be implicated and aberrantly expressed in multiple cancers. However, the pathogenic implications of LINC00518 in cervical cancer (CC) are still unclear. In this study, we focused on LINC00518 and investigated its expression pattern, clinical significance, and biological function in CC. PATIENTS AND METHODS: The expression levels of LINC00518 in CC tissues and cell lines were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), and its clinical significance was assessed by statistical analysis. Cell apoptosis was determined by flow cytometry. The cell proliferation was evaluated by MTT assay and colony forming assay, and the migration and invasion were evaluated by wound healing assays and transwell assay. Western blot was used to detect the expression of relative proteins, including EMT markers and the JAK/STAT3 signaling markers. RESULTS: We found that LINC00518 was upregulated in CC tissues and associated with International Federation of Gynaecology and Obstetrics (FIGO) stage, lymph node metastasis, depth of cervical invasion and poor survival of CC patients. Univariate and multivariate Cox regression analysis showed that LINC00518 played a significant role of independent prognostic markers in overall survival rates. Furthermore, knocking down LINC00518 expression significantly suppressed CC cell proliferation, migration and invasion, and induced apoptosis in vitro. Mechanistically, the downregulation of LINC00518 suppressed JAK/STAT3 activation and subsequently decreased N-Cadherin and Vimentin. CONCLUSIONS: The present work first suggests that LINC00518 acts as an oncogene in CC via regulation of the JAK/STAT3 signaling pathway. In the future, LINC00518 may serve as a predictive biomarker and potential therapeutic target for CC patients.

Correlation between serum P450arom and sex hormones in males with late-onset hypogonadism: a pilot study.

OBJECTIVE: The research aimed to investigate the correlation between serum P450arom and sex hormones in males with late-onset hypogonadism (LOH). PATIENTS AND METHODS: A total of 97 LOH patients and 301 matched healthy males of same age underwent androgen deficiency in the aging males (ADAM) and aging males' symptoms (AMS) scales as well as basic questionnaire survey. Serum P450arom, sex hormones, fasting blood glucose and lipid profiles were tested. General information, P450arom and sex hormone levels were compared between the LOH group and the control group. Pearson correlation analysis was used to analyze the correlation between serum P450arom concentration and AMS score, blood glucose, lipid profiles, body mass index (BMI) and sex hormones. RESULTS: Compared with the control group, the fasting blood glucose, body mass index (BMI), and Estrogen/Total Testosterone ratio (E2/TT) were significantly increased in LOH group (p<0.05), while TT, E2 and testosterone secreting index (TSI) were significantly decreased (p<0.05). No significant difference in P450arom concentration was observed between the two groups (p>0.05). The serum P450arom concentration was not related to TT, E2/TT, AMS score, and BMI. CONCLUSIONS: These findings suggested that the serum P450arom concentration is unrelated to LOH symptom score and sex hormone levels and could not be used as an observation index and diagnostic basis for LOH.

Sodium tanshinone IIA sulfate protects myocardium against paraquat-induced toxicity through activating the Nrf2 signaling pathway in rats.

OBJECTIVE:: To investigate the therapeutic effect and mechanism of sodium tanshinone IIA sulfate (STS) on paraquat (PQ)-induced myocardial injuries in a rat model. METHODS:: Healthy adult Sprague Dawley rats were randomly divided into normal control, PQ, and PQ + STS groups. PQ group was given a single intragastric administration of PQ (80 mg/kg). PQ + STS group was intraperitoneally injected with STS (1 ml/kg) at 30 min following PQ exposure. Rats in control and PQ groups were injected with equal amount of saline. After 12, 24, 48, and 72 h, rats were killed, and the apoptosis of myocardial cells was detected. Myocardial expression of Bax and Bcl-2 was measured. The activity of the nuclear erythroid 2-related factor 2 (Nrf2) pathway was assessed by Western blot. RESULTS:: The apoptotic cells in PQ group were significantly increased in a time-dependent manner compared with the control group ( p < 0.01). The rats in PQ group exhibited significantly lower Bcl-2 expression, but notably higher Bax expression at 12, 24, 48, and 72 h after PQ exposure ( p < 0.05 or 0.01). STS intervention markedly reduced the proportion of apoptotic myocardial cells, increased Bcl-2 expression, and decreased Bax expression at 24, 48, and 72 h after treatment ( p < 0.05 or 0.01). The expression of phosphorylated Nrf2 and heme oxygenase 1 in PQ + STS group was significantly increased compared with PQ and control groups ( p < 0.05 or 0.01). CONCLUSION:: STS effectively inhibits PQ-induced myocardial cell apoptosis in rats via modulating the Nrf2 pathway, suggesting its potential as a promising therapeutic agent for PQ-induced myocardium damage.

[Experience of wound treatment on extremely severe mass burn patients involved in August 2nd Kunshan factory aluminum dust explosion accident].

Objective: To explore experience of wound treatment of extremely severe mass burn patients involved in August 2nd Kunshan factory aluminum dust explosion accident. Methods: On August 2nd, 2014, 98 extremely severe burn mass patients involved in August 2nd Kunshan factory aluminum dust explosion accident were admitted to 20 hospitals in China. The patients with complete medical record were enrolled in the study and divided into microskin graft group with 56 patients and Meek skin graft group with 42 patients. Split-thickness skin in area of residual skin were resected to repair wounds of patients in microskin graft group and Meek skin graft group by microskin grafting and Meek miniature skin grafting, respectively. The residual wound size on 28 days post injury and wound infection after skin grafting of patients in the two groups, and position of donor site of all patients were retrospectively analyzed. Data were processed with t test and chi-square test. Results: The size of residual wound of patients in Meek skin graft group on 28 days post injury was (59±13)% total body surface area (TBSA), which was obviously smaller than that in microskin graft group [(70±14)%TBSA, t=4.379, P<0.05]. Twenty-nine patients in microskin graft group and 11 patients in Meek skin graft group suffered from obvious wound infection after skin grafting. Wounds of patients in two groups were repaired with residual skin around wound in head, trunk, groin, armpit, and uncommon donor sites of scrotum (4 patients), vola (10 patients), and toe or finger web (8 patients). Conclusions: Meek skin graft is the first choice for wound repair of extremely severe burn mass patients, with faster wound healing, less wound infection. Uncommon donor sites of scrotum, vola, and toe or finger web can also be used for wound repair in case of lack of skin.

MiR-16 inhibits pituitary adenoma cell proliferation via the suppression of ERK/MAPK signal pathway.

OBJECTIVE: Extracellular signal-regulated kinase (ERK)/mitogen activated protein kinase (MAPK) signal pathway participates in cell proliferation, cycle, and apoptosis. MiR-16 is down-regulated in the pituitary tumor. This study investigated the role and related mechanism of miR-16 on pituitary tumor proliferation, cycle, and apoptosis. PATIENTS AND METHODS: Dual-luciferase reporter assay was conducted to demonstrate the targeted regulation between miR-16 and MEK1. MiiR-16, MEK1, p-ERK1/2, Survivin and Cyclin D1 expression were compared between normal embryonic pituitary cells, HP75 tumor cells. Flow cytometry detection measured cell proliferation and cycle. Cultured HP75 cells were divided into four groups: miR-NC, miR-16 mimic, si-NC, and si-MEK1. Expressions of miR-16, MEK1, p-ERK1/2, Survivin, and Cyclin D1 were compared, and cell proliferation, cycle, and apoptosis were tested by flow cytometry. RESULTS: Bioinformatics analysis showed complementary binding sites between miR-16 and MEK1. Dual luciferase reporter assay validated the direct regulation between miR-16 and MEK1. Compared to that of normal pituitary tissues, significantly lower miR-16 expression, but higher MEK1 level were found in adenoma tissues. Compared to normal embryonic pituitary cells, the level of miR-16 was decreased, while the expressions of p-ERK1/2, Survivin, and Cyclin D1, along with cell proliferation or S or G2/M phase ratio were up-regulated in the group of HP75 cells. Transfection of miR-16 mimic or si-MEK1 remarkably suppressed the expressions of MEK1, p-ERK1/2, Survivin or Cyclin D1 in HP75 cells, inhibited cell proliferation and induced apoptosis and cycle arrest. CONCLUSIONS: MiR-16 inhibited ERK/MAPK pathway activity via the suppression of MEK1 expression, and further suppressed proliferation of pituitary tumor cells.

Flexibility vs Preorganization: Direct Comparison of Binding Kinetics for a Disordered Peptide and Its Exact Preorganized Analogues.

Many intrinsically disordered proteins, which are prevalent in nature, fold only upon binding their structured partner proteins. Such proteins have been hypothesized to have a kinetic advantage over their folded, preorganized analogues in binding their partner proteins. Here we determined the effects of ligand preorganization on the kon for a biomedically important system: an intrinsically disordered p53 peptide ligand and the MDM2 protein receptor. Based on direct simulations of binding pathways, computed kon values for fully disordered and preorganized p53 peptide analogues were within error of each other, indicating little if any kinetic advantage to being disordered or preorganized for binding the MDM2 protein. We also examined the effects of increasing the concentration of MDM2 on the extent to which its mechanism of binding to the p53 peptide is induced fit vs conformational selection. Results predict that the mechanism is solely induced fit if the unfolded state of the peptide is more stable than its folded state; otherwise, the mechanism shifts from being dominated by conformational selection at low MDM2 concentration to induced fit at high MDM2 concentration. Taken together, our results are relevant to any protein binding process that involves a disordered peptide of a similar length that forms a single α-helix upon binding a partner protein. Such disorder-to-helix transitions are common among protein interactions of disordered proteins and are therefore of fundamental biological interest.

Adeno-associated virus-mediated expression of myostatin propeptide improves the growth of skeletal muscle and attenuates hyperglycemia in db/db mice.

Inhibition of myostatin, a negative growth modulator for muscle, can functionally enhance muscle mass and improve glucose and fat metabolism in myostatin propeptide (MPRO) transgenic mice. This study was to investigate whether myostatin inhibition by adeno-associated virus (AAV)-mediated gene delivery of MPRO could improve muscle mass and achieve therapeutic effects on glucose regulation and lipid metabolism in the db/db mice and the mechanisms involved in that process. Eight-week-old male db/db mice were administered saline, AAV-GFP and AAV-MPRO/Fc vectors and monitored random blood glucose levels and body weight for 36 weeks. Body weight gain was not different during follow-up among the groups, but AAV-MPRO/Fc vectors resulted high level of MPRO in the blood companied by an increase in skeletal muscle mass and muscle hypertrophy. In addition, AAV-MPRO/Fc-treated db/db mice showed significantly lower blood glucose and insulin levels and significantly increased glucose tolerance and insulin sensitivity compared with the control groups (P<0.05). Moreover, these mice exhibited lower triglyceride (TG) and free fatty acid (FFA) content in the skeletal muscle, although no difference was observed in fat pad weights and serum TG and FFA levels. Finally, AAV-MPRO/Fc-treated mice had enhanced insulin signaling in the skeletal muscle. These data suggest that AAV-mediated MPRO therapy may provide an important clue for potential clinical applications to prevent type II diabetes, and these studies confirm that MPRO is a therapeutic target for type II diabetes.

Regulatory effect of estrogen receptor-α-mediated Wnt/ß-catenin signaling pathway on osteoblast proliferation.

This study was designed to investigate the regulatory effect of estrogen receptor-α (ERα)-mediated Wnt/ß-catenin signaling pathway on osteoblast proliferation. Mc3T3-El cells were infected by ERα and ERß small interfering ribose nucleic acid (siRNA) viruses and treated with estradiol 2 (E2) for 120 min after 24-h infection. Western blot was used to detect expressions of ß-catenin, Gsk 3ß, p-Gsk3ß (Ser9) and CyclinDl; and methyl thiazolyl tetrazolium (MTT) was applied to detect osteoblast proliferation after interference by different ERs. Western blot results indicated that the expressions of ß-catenin, p-Gsk3ß (Ser9) and CyclinDl decreased after ERß interference and ERα + ERß joint interference, and a more obvious decrease was found after the joint interference. After ERß interference, ß-catenin, p-Gsk3ß (Ser9) and CyclinDl were strongly expressed compared with expressions in the blank control group. MTT results demonstrated that the proliferation rate of osteoblast was lower after the joint interference than after ERß interference, while a slight increase was found in the proliferation rate after ERß interference in comparison with the blank control group. It can be concluded that estradiol is able to promote the proliferation of osteoblasts in mice by ERα-mediated Wnt/ß-catenin signaling pathway.

[Expression of PD-1/PD-L1 in lung adenocarcinoma and correlation with epidermal growth factor receptor mutation status].

Objective: To investigate the expression of programmed cell death-1(PD-1) and programmed cell death-ligand 1(PD-L1) in lung adenocarcinoma in correlation with clinical pathological parameters, especially with regard to different epidermal growth factor receptor (EGFR) mutation status. Methods: One hundred and nine cases of lung adenocarcinoma were collected during the period from Aug. 2010 to Jan. 2016, including 51 cases of EGFR wild type and 58 cases of EGFR mutations. Immunohistochemistry was used to detect PD-1/PD-L1 protein expression. Chi-square test was used to analyze the correlation between PD-1 and PD-L1 expression, and in correlation with clinicopathological parameters. All statistical analyses run by SAS 9.1 software. Results: The positive rates of PD-1 and PD-L1 expression were 68.8% (75/109) and 27.5% (30/109), respectively, with significant correlation between the two (P<0.05). PD-1 and PD-L1 expression rates were higher in 51 cases with EGFR wild type status (74.5% and 39.2%) than those in 58 EGFR mutation cases (63.8% and 17.2%); PD-1 expression was significantly associated with age (P<0.05); that of PD-L1 was closely correlated with histological type, tumor size, lymph node metastasis and EGFR status (P<0.05). Conclusions: PD-1 and PD-L1 expression profiles and their correlation with EGFR mutations are different from those with native EGFR. PD-L1 overexpression is closely correlated with larger tumor size and lymph node metastasis, suggesting it is a high-grade marker for lung adenocarcinoma.

Diabetes mellitus affects the biomechanical function of the callus and the expression of TGF-beta1 and BMP2 in an early stage of fracture healing

Transforming growth factor beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) are important regulators of bone repair and regeneration. In this study, we examined whether TGF-β1 and BMP-2 expressions were delayed during bone healing in type 1 diabetes mellitus. Tibial fractures were created in 95 diabetic and 95 control adult male Wistar rats of 10 weeks of age. At 1, 2, 3, 4, and 5 weeks after fracture induction, five rats were sacrificed from each group. The expressions of TGF-β1 and BMP2 in the fractured tibias were measured by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction, weekly for the first 5 weeks post-fracture. Mechanical parameters (bending rigidity, torsional rigidity, destruction torque) of the healing bones were also assessed at 3, 4, and 5 weeks post-fracture, after the rats were sacrificed. The bending rigidity, torsional rigidity and destruction torque of the two groups increased continuously during the healing process. The diabetes group had lower mean values for bending rigidity, torsional rigidity and destruction torque compared with the control group (P<0.05). TGF-β1 and BMP-2 expression were significantly lower (P<0.05) in the control group than in the diabetes group at postoperative weeks 1, 2, and 3. Peak levels of TGF-β1 and BMP-2 expression were delayed by 1 week in the diabetes group compared with the control group. Our results demonstrate that there was a delayed recovery in the biomechanical function of the fractured bones in diabetic rats. This delay may be associated with a delayed expression of the growth factors TGF-β1 and BMP-2.

Homology-based analysis of the GRAS gene family in tobacco.

Members of the GRAS gene family are important transcriptional regulators. In this study, 21 GRAS genes were identified from tobacco, and were classified into eight subgroups according to the classification of Arabidopsis thaliana. Here, we provide a preliminary overview of this gene family in tobacco, describing the gene structure, gene expression, protein motif organization, phylogenetic analysis, and comparative analysis in tobacco, Arabidopsis, and rice. Using the sequences of 21 GRAS genes in Arabidopsis to search against the American tobacco genome database, 21 homologous GRAS genes in tobacco were identified. Sequence analysis indicates that these GRAS proteins have five conserved domains, which is consistent with their counterparts in other plants. Phylogenetic analyses divided the GRAS gene family into eight subgroups, each of which has distinct conserved domains and biological functions. Furthermore, the expression pattern of these 21 GRAS genes reveals that most are expressed in all six tissues studied; however, some have tissue specificity. Taken together, this comprehensive analysis will provide a rich resource to assist in the study of GRAS protein functions in tobacco.

THE EFFECT OF DIHYDROARTEMISININ ON THE PROLIFERATION, METASTASIS AND APOPTOSIS OF HUMAN OSTEOSARCOMA CELLS AND ITS MECHANISM.

This study aims to research the effect of dihydroartemisinin on the proliferation, metastasis and apoptosis in human osteosarcoma cells 143B and the underlying mechanism. This study designed five groups for experiment and control, using dimethylsulfoxide (DMSO), and docosahexaenoic acid (DHA) at concentrations of 15, 25, 35 µmol.L-1 respectively. Experiments including methyl thiazolyl tetrazolium (MTT) assay, clone formation assay, Hoechst 33258 staining assay, luciferase reporter plasmid assay, Western blot and scratch test were carried out. In addition, SPSS 18.0 software from IBM was used for statistical analysis and all the data obtained from the experiments were expressed as mean ± SD, and variance was used to compare the difference between the groups. DHA is proved to be able to inhibit the proliferation and metastasis of osteosarcoma cells, as well as leaving a positive effect on apoptosis in the cytomorphosis. It achieves regulation over the human osteosarcoma cells by keeping the expression of related protein under control.

Novel homozygous deletion of segmental KAL1 and entire STS cause Kallmann syndrome and X-linked ichthyosis in a Chinese family.

Kallmann syndrome (KS) is a genetically heterogeneous disease characterised by hypogonadotrophic hypogonadism in association with anosmia or hyposmia. This condition affects 1 in 10 000 men and 1 in 50,000 women. Defects in seventeen genes including KAL1 gene contribute to the molecular basis of KS. We report the clinical characteristics, molecular causes and treatment outcome of two Chinese brothers with KS and X-linked ichthyosis. The phenotypes of the patients were characterised by bilateral cryptorchidism, unilateral renal agenesis in one patient but normal kidney development in another. The patients had low serum testosterone, follicle-stimulating hormone and luteinising hormone levels and a blunt response to the gonadotrophin-releasing hormone stimulation test. After human chorionic gonadotrophin treatment, the serum testosterone levels were normalized, and the pubic hair, penis length and testicular volumes were greatly improved in both of the patients. The two affected siblings had the same novel deletion at Xp22.3 including exons 9-14 of KAL1 gene and entire STS gene. Our study broadens the mutation spectrum in the KAL1 gene associated with KS and facilitates the genetic diagnosis and counselling for KS.

Transplantation of erythropoietin gene-transfected umbilical cord mesenchymal stem cells as a treatment for limb ischemia in rats.

The aim of this study was to investigate the mechanisms of erythropoietin (EPO)-transfected umbilical cord mesenchymal stem cells (UC-MSCs) in the treatment of rats with ischemic limb and provide a theoretical basis for optimization of UC-MSC transplantation. Sixty SD rats were randomly divided into four groups: ischemia control group, EPO treatment group, UC-MSCs treatment group, EPO gene transfected UC-MSC treatment groups (15 rats in each group). The left femoral hind artery and its branches were ligated to develop hind limb ischemia in male SD rats. Five points were injected in the adductor and gastrocnemius muscles with medium, cDNA3-EPO gene DNA-liposome complex solution or UC-MSCs in control groups and EPO-transfected-UC-MSCs in the experimental group. Western blot confirmed in vitro EPO expression in EPO gene-transfected human UC-MSCs. Arterial angiography at 4 weeks post-transplantation showed no development of blood vessels in the control group and moderate angiogenesis in the EPO- and UC-MSC-treated groups. However, a large number of freshmen angiogenesis and abundant collateral circulation was observed in the EPO-transfected-UC-MSC-treated experimental group. Rat capillary density measurement results confirmed the angiographs quantitatively and showed no statistically significant difference between EPO- and UC-MSC-treated groups (P > 0.05). CM-Dil-positive cell numbers were (0 ± 0.00), (0 ± 0.00), (32.46 ± 6.68), (59.64 ± 10.38)/HP (P < 0.05). RT-PCR detected that the in vivo mRNA expression of the EPO gene was relatively higher in the EPO-transfected-UC-MSC-treated group than the EPO-treated group (0.79 ± 0.06 vs 0.19 ± 0.04, P < 0.05). Thus, this study revealed that using UC-MSCs as vector in gene therapy for limb ischemia facilitates stable and effective expression of EPO compared to direct gene injection.