ABSTRACT
Objective To explore the effect of overexpression of retinoic acid receptor α(RARα)on epithelial-to-mesenchymal transition(EMT)induced by hypoxia in renal tubular epithelial cells(NRK-52E).Methods The RARα lentivirus vector and negative control lentivirus vector were synthetised.The NRK-52E cells were divided into 4 groups:the normal control group,the hypoxia model group,the transfection group and the negative control group.Puro-mycin(2 mg/L)was added in transfection group and negative control group for screening after gene interference for 72 h.Then the 2 groups were subjected to hypoxia/reoxygenation,but the normal control group had no treatment. The change of cellular morphology was observed by using light microscope;the mRNA and protein expressions of RARα, E-cadherin,α -smooth muscle actin(α-SMA)in NRK-52E cells were detected by adopting reverse transcription-polymerase chain reaction(RT-PCR)and Western blot after hypoxia for 48 h.Results (1)Light microscope re-vealed that cells in both hypoxia model group and negative control group cells became atrophic and elongated,which were consistent with the morphology of myofibroblasts.But cells in transfection group cells were cubic,forming an epi-thelial monolayer.(2)Compared with the normal control group,the mRNA and protein expressions of RARα and E-cadherin in hypoxia model group were dramatically reduced(mRNA:0.58 ± 0.12 vs.1.00 ± 0.00,0.11 ± 0.00 vs. 1.00 ± 0.00,t= -0.63,767.30,all P<0.05;protein:0.63 ± 0.12 vs.1.62 ± 0.16,0.44 ± 0.22 vs.1.27 ± 0.08,t=8.61,6.19,all P<0.05),but the mRNA and protein expressions of α-SMA were higher(3.47 ± 0.83 vs.1.00 ± 0.00,1.39 ± 0.16 vs.0.64 ± 0.10,t= -5.01,-6.91,all P<0.05).(3)The mRNA and protein expressions of RARα and E-cadherin in the transfection group were significantly increased,compared with hypoxia model group(mRNA:4.69 ± 1.34 vs.0.58 ± 0.12,0.23 ± 0.00 vs.0.11 ± 0.00,q=9.13,25.48,all P<0.05;protein:1.39 ± 0.19 vs. 0.63 ± 0.12,0.87 ± 0.09 vs.0.44 ± 0.22,q=7.92,4.30,all P<0.05)and negative control group(mRNA:4.69 ± 1.34 vs.0.55 ± 0.21,0.23 ± 0.00 vs.0.12 ± 0.01,q=9.20,23.35,all P<0.05;protein:1.39 ± 0.19 vs.0.65 ±0.18,0.87 ± 0.09 vs.0.39 ± 0.21,q=7.71,4.80,all P<0.05).Conversely,the mRNA and protein levels of α-SMA were obviously lower in transfection group(1.52 ± 0.34 vs.3.47 ± 0.83,4.05 ± 0.81,0.82 ± 0.13 vs.1.39 ± 0.10,1.17 ± 0.10,q=4.88,6.33,7.50,4.61,all P<0.05).The difference in mRNA and protein expressions of RARα,E-cadherin,α-SMA between the hypoxia group and the negative control group had no statistical significance (all P>0.05).Conclusion Overexpression of RARα could alleviate EMT of renal tubular epithelial cells induced by hypoxia.
ABSTRACT
Objective To explore the role of c-Jun N-terminal protein kinase(JNK)pathway in renal tubular epithelial cells(RTEC)transdifferentiation by detecting the expression of JNK pathway in the injury of hypoxic RTEC. Methods In vitro cultured rat RTEC were randomly divided into four groups of normal control, hypoxia,inhibitors,dimethyl sulfoxide(DMSO). The groups of hypoxia,inhibitor and DMSO were placed into a vacuum tank to establish hypoxia model. The cells were harvested 6,12 and 24 h after hypoxia started. RT-PCR and Western blot test were used to detect the mRNA expressions of alpha smooth muscle actin(α- SMA)and protein expressions of JNK,pJNK and alpha SMA,respectively,in all the groups. Results Hypoxia induced significantly increased expressions of α-SMA mRNA and proteins,and JNK and pJNK proteins as well in RTEC (all P<0.05).After addition of inhibitors,the expression of α-SMA mRNA and proteins,the protein expression of pJNK in RTEC were significantly decreased(all P < 0.05)and the expression of JNK protein was significantly increased(P < 0.05). There was no significant difference in the expressions of JNK,pJNK,α-SMA in RTECs between hypoxia group and DMSO group(all P>0.05).There was a significant positive correlation between pJNK protein and α-SMA protein expression(P < 0.01). Conclusion In hypoxic RETC injury in rats,JNK pathway may be involved in the RTEC phenotype transdifferentiation.
ABSTRACT
<p><b>OBJECTIVE</b>To detect disease-causing mutations in a patient with hereditary elliptocytosis.</p><p><b>METHODS</b>Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) was used to identify the type of erythrocyte membrane protein defect. Potential mutations of the exons and adjacent introns of relevant genes were analyzed by Sanger sequencing.</p><p><b>RESULTS</b>SDS-PAGE has failed to detect any difference between the patient and healthy controls. However, Sanger sequencing has detected three mutations in the SPTA1 gene in the patient, which included c.5077A>C (p.Lys1693Gln) missense mutation in exon 36, c.5572C>G (p.Leu1858Val) missense mutation in exon 40, and a IVS45nt-12C>T in intron 45. The father and grandmother of the patient were both heterozygous for c.5077A>C mutation, while her mother was heterozygous for c.5572C>G and IVS45nt-12C>T mutations.</p><p><b>CONCLUSION</b>The hereditary elliptocytosis in the patient may be attributed to the synergistic action of c.5077A>C, c.5572C>G and IVS45nt-12C>T mutations of the SPTA1 gene.</p>
ABSTRACT
BACKGROUND:Shape-memory polymer materials can be used as intrauterine device materials, which have certain shape changes under the influence of external temperature, and have good biodegradation, and are harmless to human body. OBJECTIVE:To prepare a poly(D,L-lactide)-based shape-memory intrauterine device and to analyze its application features. METHODS:Poly(D,L-lactide)-based shape-memory intrauterine device was prepared in this experiment. Sixty female New Zealand rabbits were randomly divided into two groups, with 30 rabbits in each group: experimental group were bilateraly implanted poly(D,L-lactide)-based shape-memory intrauterine device, and control group had no intrauterine device. Gross observation, blood test and histological examination were conducted at different time after implantation. RESULTS AND CONCLUSION:After 4 weeks of implantation, the intrauterine device was smooth and thickened and shortened compared with before implantation; after 6 weeks of implantation, the device became thinned and further shortened and even disappeared partialy. No significant difference was found in the leukocyte, erythrocyte, hemoglobin, platelet, creatinine and urea levels between the two groups at 6 weeks after implantation. In the experimental group, obvious edema and hyperemia in the tubal mucosa were visible at 4 weeks after implantation, and a very smal amount of infiltrated lymphocytes were present at 6 weeks after implantation, and edema and hyperemia were improved significantly. These findings indicate that after implantation, the poly(D,L-lactide)-based shape-memory intrauterine device can deform and degrade, and result in a certain pressure on the oviduct wal, but has no cel toxicity and possesses good application security.
ABSTRACT
A total of 130 anvolatory infertile outpatients were divided randomly into 4 groups of LE1 (letrozole 2.5 mg,n =45),LE2 (letrozole 5 mg,n =30),CC1 (clomiphene citrate 50 mg,n =35) and CC2 (clomiphene citrate 100 mg,n =20).The treatments started at day 5 of menstruation and lasted for 5 days.The size and number of ovarian follicles and thickness of endometrium of all 4 groups were measured starting from day 10 of menstruation.8 000-10 000 units of human chorionic gonadotropin (hCG) was injected when ovary follicles were mature.And blood estradiol (E2) was measured at the same day.Ovulation,pregnancy and adverse reaction were also monitored.The pregnancy rate was 33％ (n =15) in group LE1 and 14％ (n =5) in group CC1 with significant statistical difference (x2 =4.00,P ＜ 0.05).And there were 43 successful ovulation periods in 55 periods with an ovulation rate of 78％ in group LE1;28 successful ovulation periods in 52 periods with an ovulation rate of 54％ in group CC with significant statistical difference (X2 =4.60,P ＜0.05).E2 level (pmol/L) in the day of hCG injection in group LE1 was (774.2 ± 126.0) pmol/L and in group CC1 (963.7 ± 108.3) pmol/L,t =3.98 with significant difference (P ＜ 0.01).The number of follicle in group LE1 was 1.20 ± 0.38;CC1 2.28 ± 0.88,t =4.76 with significant difference (P ＜0.01).The thickness of endometrium in the day of hCG in LE1 was (9.6 ± 1.3) mm;CC1 (5.9 ± 1.1) mm,t =6.20 with significant difference (P ＜ 0.01).No significant statistical differences existed between LE1 and LE2,CC1 and CC2 in pregnancy rate,successful ovulation,number of mature follicles,E2 level and thickness of endometrium at the day of hCG injection.The outcomes of 36 pregnancies were 32 full-term deliveries with neonatal health,2 early abortions and 2 prematures.Mild ovarian hyperstimulation syndrome (OHSS) had a low occurrence rate.
ABSTRACT
A total of 104 patients with cervical squamous cell cancer (Figo Ⅰ b2-Ⅱ b) were recruited during March 2006 and October 2011.One group (A,n =50) received preoperative intravenous neoadjuvant chemotherapy while another group (B,n =54) had interventional arterial chemotherapy plus bilateral uterine artery embolization.Surgery was performed for those with a change of cancer size either as complete or partial remission at Week 2 post-chemotherapy according to the WHO evaluation criteria.Otherwise radiotherapy was implemented.The ratio of surgery after chemotherapy was 70％ (n =35) in Group A versus 87％ (n =47) in Group B (P ＜ 0.05).For Group A,there were blood vessel invasion (n =6,17％),lymph node metastasis (n =8,23％) and positive margin (n =1,3％).For Group B,(n =7,15％),(n =9,21％) and (n =1,2％).There was no significant inter-group statistic difference (P ＞ 0.05).Preoperative chemotherapy is effective for cervical squamous cancer (tumor diameter ＞ 4 cm)with fewer side effects.