[mRNA expression of MDR3 gene in the blood of preterm infants with parenteral nutrition-associated cholestasis].

OBJECTIVE: To study the association between the expression of the MDR3 gene and the pathogenesis of parenteral nutrition-associated cholestasis (PNAC) in preterm infants. METHODS: Among the preterm infants who were admitted to the hospital from June 2011 to November 2017 and received parenteral nutrition for more than 14 days, 80 who did not develop PNAC were enrolled as non-PNAC group, and 76 who developed PNAC were enrolled as PNAC group. On days 1, 14, 30, 60 and 90 after birth, serum hepatobiliary biochemical parameters [alanine aminotransferase (ALT), total bilirubin (TBil), direct bilirubin (DBil), total bile acid (TBA) and gamma-glutamyl transpeptidase (γ-GT)], fibrosis indices [hyaluronic acid, laminin, procollagen III N-terminal peptide and type IV collagen] and clinical manifestations were observed. Real-time quantitative PCR was used to measure the mRNA expression of MDR3 in both groups, and the correlation between the mRNA expression of MDR3 and serum hepatobiliary biochemical parameters was analyzed. RESULTS: In the PNAC group, serum levels of hepatobiliary biochemical parameters and fibrosis indices increased on day 14 after birth and reached the peak on day 30 after birth, followed by a reduction on day 60 after birth. On days 14, 30, 60 and 90 after birth, the PNAC group had significantly higher serum levels of hepatobiliary biochemical parameters and fibrosis indices than the non-PNAC group (P<0.05). The PNAC group had higher relative mRNA expression of MDR3 in peripheral blood cells than the non-PNAC group (P<0.05). In the PNAC group, the relative mRNA expression of MDR3 in peripheral blood cells was negatively correlated with serum levels of hepatobiliary biochemical parameters (ALT, TBil, DBil, TBA and γ-GT) (P<0.001). CONCLUSIONS: High mRNA expression of MDR3 in preterm infants may be associated with the development of PNAC, and further studies are needed to identify the mechanism.

Effects of Prenatal PM10 Exposure on Fetal Cardiovascular Malformations in Fuzhou, China: A Retrospective Case-Control Study.

BACKGROUND: Maternal exposure to ambient air pollution has been associated with an increased risk of congenital heart defects in offspring; however, the results are inconsistent. OBJECTIVES: We investigated whether there is an association between prenatal exposure to particulate matter with diameter ≤10µm (PM10) during early pregnancy and fetal cardiovascular malformations. METHODS: The gravidae from a hospital-based case­control study in Fuzhou, China, during 2007­2013 were assigned 10-d or 1-mo averages of daily PM10 using an air monitor­based inverse distance weighting method during early pregnancy. A total of 662 live-birth or selectively terminated cases and 3,972 live-birth controls were enrolled. The exposure was considered as a categorical variable. A multivariable logistic regression model was constructed to quantify the adjusted odds ratios (aORs) of the exposure to PM10 and the risks of fetal cardiovascular malformations. RESULTS: PM10 levels were positively associated with the risks of atrial septal defect (aORs ranging from 1.29 to 2.17), patent ductus arteriosus [aORs = 1.54, 1.63; 95% confidence intervals (CIs): 1.17, 2.23; 1.06, 3.24], overall fetal cardiovascular malformations (aOR = 1.28; 95% CI: 1.03, 1.61), ventricular septal defect (aOR = 1.19; 95% CI: 1.00, 1.43), and tetralogy of Fallot (aOR = 1.44; 95% CI: 1.01, 2.19) in the various observed periods scaled by 10 d or 1 mo in the first and second gestation months. The strongest associations were observed for exposure to PM10 in the second quartile, whereas the associations were attenuated when higher concentrations of PM10 in the third and fourth quartiles of the exposure were evaluated. No correlations of PM10 levels with these cardiovascular malformations in the other time periods of gestation were observed. CONCLUSIONS: Our findings suggest some positive associations between maternal exposure to ambient PM10 during the first two months of pregnancy and fetal cardiovascular malformations. https://doi.org/10.1289/EHP289.

Relationship between resistin and IL-23 levels in follicular fluid in infertile patients with endometriosis undergoing IVF-ET.

BACKGROUND: Endometriosis (EM) interferes with the reproductive process and affects the success rate of in vitro fertilization (IVF). Inflammatory cytokines are suggested to play a role in infertility in patients with EM. OBJECTIVES: In this study, we aimed to investigate the relationship between resistin and interleukin 23 (IL-23) levels in follicular fluid (FF) and serum together with the severity of endometriosis and in vitro fertilization/ embryo transfer (IVF-ET) outcome. MATERIAL AND METHODS: Samples from 116 infertile women were studied using enzyme-linked immunosorbent assay (ELISA). The study group consisted of 76 infertile patients diagnosed with EM (40 with stages I-II and 36 with stages III-IV) undergoing IVF-ET. The control group included 40 women with tubal factor infertility. FF and serum samples were collected on the day of follicle aspiration and hCG administration, respectively. RESULTS: The serum and FF resistin levels were significantly higher in the EM group than in the control group (p-value <0.05). The FF resistin and IL-23 levels were significantly higher in EM stages III-IV than in stages I-II (p-value <0.05), and the serum resistin and IL-23 levels were also significantly (p-value <0.01) higher in stages III-IV than in stages I-II. The E2 level on the day of hCG administration and the implantation rate were both significantly lower in the EM group than in the control group. However, there were no differences in the Gn duration and dose, and the cleavage, implantation and clinical pregnancy rates between the 2 groups. CONCLUSIONS: Our results suggest that patients with EM exhibit increased resistin level in FF and serum. Advanced EM may contribute to infertility via decreased embryo implantation rates because of inflammation and immune rejection. No influence was observed on pregnancy outcomes after IVF-ET.

Griffipavixanthone from Garcinia oblongifolia champ induces cell apoptosis in human non-small-cell lung cancer H520 cells in vitro.

Griffipavixanthone (GPX) is a dimeric xanthone which was isolated in a systematic investigation of Garcinia oblongifolia Champ. In this study, we investigate the effect of GPX on cell proliferation and apoptosis on human Non-small-cell lung cancer (NSCLC) cells in vitro and determine the mechanisms of its action. GPX inhibited the growth of H520 cells in dose- and time-dependent manners, with IC50 values of 3.03 ± 0.21 µM at 48 h. The morphologic characteristics of apoptosis and apoptotic bodies were observed by fluorescence microscope and transmission electron microscope. In addition, Annexin V/PI double staining assay revealed that cells in early stage of apoptosis were significantly increased upon GPX treatment dose-dependently. Rh123 staining assay indicated that GPX reduced the mitochondrial membrane potential. DCFH-DA staining revealed that intracellular ROS increased with GPX treatment. Moreover, GPX cleaved and activated caspase-3. In summary, this study showed that GPX inhibited H520 cell proliferation in dose- and time-dependent manner. Further mechanistic study indicated that GPX induced cell apoptosis through mitochondrial apoptotic pathway accompanying with ROS production. Our results demonstrate the potential application of GPX as an anti-non-small cell lung cancer agent.

MicroRNA-199a-3p regulates endometrial cancer cell proliferation by targeting mammalian target of rapamycin (mTOR).

BACKGROUND: Abnormal expression of miR-199a-3p, which has similar effects to oncogenes or tumor suppressor genes, can occur in various malignant tumors and is closely linked with tumor cell proliferation, invasion, and metastasis. However, its expression and effects in endometrial endometrioid adenocarcinoma (EEC) are still unclear. This study was designed to identify the impact of miR-199a-3p on the proliferation of EEC cells and its role in the carcinogenesis of EEC. METHODS: The expression levels of miR-199a-3p in EEC and paired adjacent nontumor tissues were analyzed by real-time polymerase chain reaction. The effects of miR-199a-3p on proliferation, cell cycle and apoptosis in EEC cells were analyzed in Ishikawa cells transfected with miR-199a-3p mimics and inhibitors. The target genes of miR-199a-3p were predicted using bioinformatics methods. The extent of regulation of the predicted target genes by miR-199a-3p was determined using luciferase reporter assays, Western blotting, and quantitative polymerase chain reaction. The EEC cells were pretreated with target gene-specific inhibitors to further identify the relationship between the effects of miR-199a-3p and the predicted target genes. RESULTS: Compared with the adjacent tissues and normal endometrium, reduced expression of miR-199a-3p was found in human EEC specimens. Compared with the control group transfected with control microRNA mimics, the proliferative capacity of EEC cells transfected with miR-199a-3p mimics was inhibited, whereas cells transfected with miR-199a-3p inhibitors showed increased proliferation. The inhibitory effect was associated with increased cell populations at the G1-phase, and decreased cell populations at the S-phase. The results demonstrated that miR-199a-3p could inhibit the protein expression of mammalian target of rapamycin (mTOR) by targeted binding to the mTOR-3' untranslated region. Inhibition of EEC cell proliferation by miR-199a-3p was mediated by its targeted regulation of mTOR. CONCLUSIONS: MiR-199a-3p inhibits tumor cell proliferation through negative regulation of mTOR expression. Restoration of intracellular miR-199a-3p levels may serve as a potential option for EEC treatment.

[Study on modified Prolift for pelvic floor reconstruction in the prevention of stress urinary incontinence].

OBJECTIVE: To evaluate the safety and efficacy of modified Prolift pelvic floor reconstruction with improving the placement of Prolift-A in treatment of severe pelvic floor dysfunction and stress urinary incontinence (SUI). METHODS: From July 2008 to September 2010, 170 cases with severe pelvic organ prolapse (POP) treated by modified Prolift pelvic floor reconstruction surgery in Fuzhou General Hospital were enrolled in this study. The Prolift-A was laid tension-free under the mid-urethra with the position of Prolift-A displaced from the neck of bladder to the mid-urethra. No concomitant tension-free urethra suspender via vagina was performed. Primary outcomes were assessed with POP quantitation (POP-Q) system to evaluate the postoperative anatomical replacement stage. Secondary outcome measure were: urogenital distress inventory 6 (UDI-6), the incontinence impact questionnaire 7 (IIQ-7) and the pelvic floor incontinence questionnaire 7 (PFIQ-7) to evaluate the impact on life quality at the follow-up of 1, 6, 12 months. RESULTS: At 6 and 12 months after surgery, 168 cases and 163 cases were followed up. The anatomical cure rates were 98.8% (166/168) at 6 months and 97.5% (159/163) at 12 months, respectively. One case with bladder injury and 1 case with rectum injury were observed. Five cases with recurrence were observed, including 2 cases with anterior vagina prolapse, 2 cases with uterine prolapse and 1 case with posterior vagina prolapse. Meanwhile, 3 cases with hematoma and 7 cases with mesh erosion were observed. Quality of life of all patients were improved significantly by UDI-6, IIQ-7 and PFIQ-7 scoring system evaluation. Among 79 POP patients with SUI, the cure rate of SUI was 93.7% (74/79). Of 5 cases with symptomatic SUI, 2 cases were needed surgical intervention. Twenty-three cases were found with minimal SUI symptoms and subjective satisfaction without objective influence on quality of life. Seven patients presented dysuria after surgery, 5 cases recovered urination with 10 days, 1 case recovered with 1 months, and 1 case with 6 months by bladder drainage. Eleven cases with discomfort urination and 3 cases with slow urination were found. CONCLUSIONS: The modified Prolift pelvic reconstructive surgery was safe and efficacy intervention in treatment of POP and prevention of SUI.

Two new lignans from the stems of Schisandra bicolor.

Two new lignans, named zuihonins E (1) and F (2), were isolated from the stems of Schisandra bicolor Cheng var. tuberculata Law. The structures of the new lignans were elucidated on the basis of extensive spectroscopic analysis, including 1D, 2D NMR, and MS experiments, and their absolute stereochemistry was determined by circular dichroism spectrum. Compounds 1 and 2 did not inhibit the growth of hepatoma carcinoma cell (HepG2), lung carcinoma cell (A549), and human breast carcinoma (MCF-7) cell lines.

Occurrence of nonylphenol and nonylphenol monoethoxylate in soil and vegetables from vegetable farms in the Pearl River Delta, South China.

Low molecular-mass nonylphenol ethoxylates (NPEOs) and 4-nonylphenol (NP) are biodegradation products of higher molecular mass NPEOs used as surface active agents, and they are endocrine-disrupting contaminants. In this study, surface soil (0-20 cm) samples and different vegetable samples were collected from 27 representative vegetable farms located in Shenzhen, Dongguan, and Huizhou within the Pearl River Delta region, South China, and NP and nonylphenol monoethoxylate (NP(1)EO) were analyzed using high-performance liquid chromatography with ultraviolet detection. The results show that NP and NP(1)EO were detected in soil and vegetable samples. The concentrations of NP and NP(1)EO in soil samples ranged from nondetectable (ND) to 7.22 µg kg(-1) dry weight (dw) and from ND to 8.24 µg kg(-1) dw, respectively. The average concentrations of both NP and NP(1)EO in soil samples decreased in the following order: Dongguan > Huizhou > Shenzhen. The levels of NP and NP(1)EO in vegetable samples varied from 1.11 to 4.73 µg kg(-1) dw and from 1.32 to 5.33 µg kg(-1) dw, respectively. The greatest levels of both NP and NP(1)EO were observed in water spinach, and the lowest levels of NP and NP(1)EO were recorded in cowpea. The bioconcentration factors (the ratio of contaminant concentration in plant tissue to soil concentration) of NP and NP(1)EO were <1.0 (mean 0.535 and 0.550, respectively). The occurrences of NP and NP(1)EO in this study are compared with other studies, and their potential sources are discussed.

[Prenatal molecular diagnosis of four fetuses at high risk for X-linked adrenoleukodystrophy].

OBJECTIVE: To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy (X-ALD). METHODS: The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells. Maternal contamination was evaluated by paternity test. PCR-RFLP, sequencing and denaturing high performance liquid chromatography (DHPLC) were used to detect the ABCD1 gene of fetal genome. RESULTS: In the pedigree 1, the PCR product (799 bp) of the fetus 1 and her father (normal control) could be digested with BcnI. No P560L mutation, which was present in the index patient, was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing. In the pedigree 2, the PCR product (232 bp) of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation, which was present in the propositus, was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing. In the pedigree 3, the PCR product (271 bp) was digested with AciI, the pattern of the fetus 3 and the propositus being the same, and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing. In the pedigree 4, the PCR product (269 bp) was analyzed with the DHPLC, and the pattern of elution peaks of the fetus 4 and her father was similar, but different from that of the propositus. No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing. Judging from the sex of the fetuses, fetuses 1 and 2 were normal homozygotes, fetus 3 was an ALD hemizygote, and fetus 4 was a normal hemizygote. CONCLUSION: A new protocol for X-ALD prenatal molecular diagnosis is proposed, which would ensure the accuracy of prenatal diagnosis.

[Prenatal diagnosis of 5 fetuses with high risk of developing spinal muscular atrophy].

OBJECTIVE: To perform prenatal diagnosis for 5 pregnant women who had given birth to children with spinal muscular atrophy (SMA). METHODS: Thirty to forty mililiters of amniotic fluid was obtained by amniocentesis under ultrasonic monitoring. DNA was extracted directly from sediment of amniotic fluid. Short tandem repeat (STR) profiling was carried out to evaluate the contamination of amniotic DNA by maternal genomic DNA. Two methods, PCR-restriction fragment length polymorphism (PCR-RFLP) and allele-specific PCR, were used to analyze exon 7 of SMN gene from amniotic DNA. RESULTS: Comparing the 16 STR sites of each fetus with those of his/her parents, there was no or little contamination of amniotic DNA by maternal genomic DNA. In conventional PCR-RFLP, part of the PCR product (189 bp) from amniotic DNA of fetus A, C, or D remained intact after digestion with Dra I, while the PCR product from amniotic DNA of fetus B or E was completely digested by Dra I. In allele-specific PCR, exon 7 of both SMN1 and SMN2 gene could be seen when amniotic DNA of fetuses A, C, or D was analyzed, while only exon 7 of SMN2 could be seen when amniotic DNA of fetuses B or E was analyzed. CONCLUSION: Homozygous deletion of SMN1 is not detected in fetuses A, C, and D, predicting that the risk of developing SMA after birth would be extremely low. Homozygous deletion of SMN1 was present in fetuses B and E suggesting high risk of developing SMA after birth.

Prenatal molecular diagnosis of adrenoleukodystrophy.

OBJECTIVE: To carry out prenatal diagnosis on two fetuses of different pedigrees with X-linked adrenoleukodystrophy (ALD). METHODS: The amniotic fluid was obtained with the help of a clinical doctor and the genomic DNA was isolated from it. Maternal DNA contamination was excluded by fluorescent STR profiling, The R617G mutation found in the first pedigree was searched in genomic DNA of amniotic fluid cells (AFC) from fetus 1 by amplification refractory mutation system (ARMS) and dot DNA hybridization while the P534R mutation found in pedigree 2 was analyzed in the AFC genomic DNA of fetus 2 by restrictive digestion with Hae II and DNA direct sequencing. RESULTS: A specific band (185 bp) was detected from the genomic DNA of the first fetus and his mother by using mutation primer in ARMS but not from that of the first fetus's father and unrelated controls. DNA dots were visualized only in the fetus 1 and carrier when using the mutation probe in DNA hybridization. In the other ALD family, the PCR product (506 bp) of the second fetus which spanned the site of P534R mutation could not be digested with Hae II and no mutation was detected in the ABCD1 gene from the genomic DNA of the fetus 2 by using DNA direct sequencing. CONCLUSION: Fetus 1 had R617G mutation on his ABCD1 gene and he was an adrenoleukodystrophy hemizygote. Fetus 2 had no P534R mutation on his ABCD1 gene and he was a normal hemizygote.

[Clinical analysis of different surgeries for women with stress urinary incontinence].

OBJECTIVE: To compare the efficacy of four kinds of suspender surgery: tension-free vaginal tape (TVT), trans-obturator tape (TOT), Goebell-Stoeckel fascia lata sling (Lata) and Burch urethropexy (Burch) in the treatment of stress urinary incontinence. METHOD: In this controlled, retrospective study, 103 patients were enrolled, including 53 patients who were operated on by TVT, 16 patients by TOT, 19 patients by Lata and 15 patients by Burch. RESULTS: The cure rate was as follows: TVT 94% (50/53), TOT 94% (15/16), Lata 95% (18/19), and Burch 87% (13/15) three months after the operation. The operative time for TVT, TOT, Lata and Burch was (28 +/- 7), (16 +/- 5), (125 +/- 13), and (43 +/- 6) min, whereas the duration of postoperative catheterization was (26 +/- 3), (3 +/- 1), (120 +/- 6), and (72 +/- 5) h, respectively. Two cases in TVT group had bladder perforation, one case in TVT group and two cases in Lata group had urinary retention postoperatively. CONCLUSIONS: The efficacy of the four kinds of suspender surgery is obvious. TVT and TOT have the advantage of minimal invasion, quick recovery, and short duration of hospitalization and catheterization, with the possibility of accompanying operations of uterine and vaginal prolapse at the same time. Burch can also be operated on concomitantly with other gynecological operations.

[Mutational analysis of three Chinese pedigrees with adrenoleukodystrophy].

OBJECTIVE: To identify the mutational genotype of three Chinese families with X-linked adrenoleukodystrophy (X-ALD: MIM#300100). METHODS: Total RNA was extracted from the peripheral blood leukocytes of patients 1, 2 and the mother of patient 3, using RNA blood Mini kit (QIAGEN). After reverse transcription, cDNA was amplified in four overlapping segments. The PCR products were purified and directly sequenced. To confirm the mutations, the genomic DNA was isolated from the patients and their family members using DNA blood isolation kit (MO-BIO) and analyzed by PCR-restrictive digestion or amplification refractory mutation system. RESULTS: Three distinct mutations were detected in the ABCD1 gene of the three pedigrees. A mutation of CCC-->CGC was detected at codon 534 of the ABCD1 gene from patient 1, resulting in the arginine for proline substitution. A change of GGG-->AGG was found at codon 266 of the second patient's gene, accompanied with the replacement of glycine by arginine. A mutation of CGC-->GGC was found at codon 617 in one ABCD1 allele of the third patient's mother, leading to the glycine for arginine substitution. The three mutations were confirmed through restriction analysis or amplification refractory mutation system. CONCLUSION: Three ABCD1 gene missense mutations were detected in three unrelated Chinese families with X-linked adrenoleukodystrophy, one of which, the mutation (P534R), is novel in Chinese with ALD, and the other two G266R and R617G mutations, have been reported outside China.

[Comparative study of tension-free vaginal tape and fascia lata for stress urinary incontinence].

OBJECTIVE: To compare the efficacy of surgically managing stress urinary incontinence (SUI) with tension-free vaginal tape (TVT) and autologous fascia lata pubovaginal sling (Lata). METHODS: Sixty-seven patients suffering from SUI entered this prospective study. The patients were divided into two groups randomly, 48 in TVT group and 19 in Lata group. The average follow-up time post-operation was 20 months in TVT group and 37 months in Lata group. RESULTS: The operating time was (27 +/- 5) min in TVT and (125 +/- 13) min in Lata. The volume of blood loss during surgery was: (27 +/- 6) ml in TVT and (67 +/- 11) ml in Lata. The post-operative recovery period in the hospital was: TVT, 1.8 days and Lata, 7.2 days. The differences between the two groups for these three parameters were statistically significant (P < 0.01). The 3-month cure rate post-operations was 94% (45/48) for TVT and 95% (18/19) for Lata. The surgical complications included 1 case of vesical perforation and 2 cases of overactive bladder in TVT group. CONCLUSIONS: TVT and Lata are both effective and safe surgical treatments for stress urinary incontinence. TVT has the benefits of being minimally invasive with a shorter operating time and a quicker recovery for the patients. A disadvantage is that the tape used in TVT is expensive, making the operation rather costly. Lata on the other hand is cheaper and may result in fewer complications, but the operation is not as easy for the patient.

[Treatment of the Goebell-Stoeckel fascia lata sling operation for female stress urinary incontinence].

OBJECTIVE: To observe the efficacy and safety of the goebell-stoeckel fascia lata sling operation for female stress urinary incontinence. METHODS: Thirteen patients suffering from stress urinary incontinence (SUI) with prolapsed vagina were enrolled into this study. The average follow-up time was thirty months. The outcomes and complications after operation were analysed. RESULTS: All patients had complete cure. There were no vaginal bleeding, infection, rejection for surgery and no recurrent cases. Five patients had mild temporary dysuria, which disappeared within one month. CONCLUSION: The goebell-stoeckel fascia lata sling operation is an effective, safe, well-tolerated procedure for treatment of SUI.