Objective To detect the expression of miRNA -34a in normal cervical,low grade cervical intra-epithelial neoplasia and high grade cervical intraepithelial neoplasia,to explore its clinical significance.Methods The expression levels of miRNA -34a in 82 cases of low grade cervical intraepithelial neoplasia (CINⅠ,CIN L group),87 cases of high grade cervical intraepithelial neoplasia (CINⅡ -Ⅲ,CIN H group) and 60 cases of normal cervical tissues (control group) in tissue specimens and serum were examined by real -time PCR(RT -PCR). Results The relative expression levels of miRNA -34a in tissues and serum of CIN H group were (0.584 ± 0.288),(0.528 ±0.299),which of CIN L group were (0.903 ±0.252),(0.847 ±0.279),which of the control group were (0.960 ±0.212),(0.908 ±0.223).The miRNA -34a level of CIN H group in tissues and serum were lower than CIN L group and control group,there was significant difference( Ptissues =0.001,Pserum =0.003).The miRNA -34a level in tissue and serum had no significant difference between the CINL group and control group(P >0.05).The expression level of serum miRNA -34a was positively related with the level of the tissues(r =0.907,P <0.01).Conclusion The expression level of miRNA -34a in high grade CIN was significantly decreased,which indi-cated that miRNA -34a play an important role in the prognosis of CIN and the transformation of cervical cancer,and it has certain clinical value for the diagnosis and treatment of CIN.

Case discussion is an effective approach,which combines basic theory with clinical medicine.It can evoke students' interest and cultivate their creative thinking capacity.Moreover,it can improve teachers' general ability in teaching pathophysiolo- gy.In this article,we discuss the application of case discussion in pathophysiology teaching.

Objective To investigate the role of muhiplex ligation-dependent probe amplification(MLPA) in identifying fetal aneuploidy of chromosomes 13,18,21,X,and Y. Methods From June 2007 to December 2008,263 samples(prenatal diagnosis group),including amniotic or umbilical cord blood from pregnant women who required prenatal diagnosis,were processed in parallel by MLPA and conventional karyotype to detect fetal aneuploidy.Another 26 samples(fetal death group).ineluding retained abortion and fetal death,were also processed bv MLPA. Results Five cases of 21-trisomy,4 eases of 18-trisomy,1 case of 13-trisomy and 3 cases of 45,X were identified among the prenatal diagnosis group by MLPA,and the results were consistent with karyotype.Two cases of 45,X and 1 case of 18-trisomy were identified among the retained abortion and fetal death group. Conclusions MLPA is a rapid,efficient,simple,reliable and economical technique in detecting most common chromosomal aneuploidies and have important clinical value.

Studies have suggested that human cytomegalovirus-activated infection is closely associated with atherosclerosis. The levels of interleukin-8 increase significantly in human cyto-megalovirus infection-related atherosclerosis, inducing and aggravating inflammatory reaction through the chemokine receptors, and thus plays an important role in the process of atherosclerosis.

Objective To explore the relationship between Stathmin-1 and human papilloma viruse (HPV) persistent infection after conization of uterine cervix, and to show the clinical significance to recurrent of cervical intraepithelial neoplasia (CIN). Methods One hundred and six patients who were treated with conization of uterine cervix for CIN 2-3 grades were enrolled. Thirty-six recurrent patients were enrolled in recurrence group, and the others were enrolled in control group. The expression of Stathmin-1 in primary CIN tissues in two groups was detected by immunohistochemistry. The HPV infection was detected by HPV-DNA test. The relationship of HPV persistent infection and recurrence was analyzed. Results The positive expression rate of HPV persistent infection and HPV persistent infection rate in recurrence group were 88.89%(32/36), 83.33%(30/36), in control group were 34.29%(24/70) and 22.86%(16/70), and there were significant difference (P <0.01). Forty-two patients had HPV persistent infection in 56 patients with stathmin-1 positive expression, and 4 patients had HPV persistent infection in 46 patients Stathmin-1 negative expression. There was positive correlation (r=0.97, P<0.01). The type of HPV persistent infection in two groups was no significant difference (P >0.05). Conclusions Stathmin-1 positive expression is related to HPV persistent infection. The two factors can affect the prognosis of high-grade CIN, and can provide new cues and theory basis for the prevention of recurrence.

Objective:To prove whether La protein could effect the replications of the HBV in cultured cells.Methods:Three specific SiRNAs for human La protein was obtained by transcription in vitro. After transfection with the SiRNAs into HepG2.2.15 cell, the levels of La protein mRNA and HBV DNA were detected by real-time PCR.Results:The HBV DNA secreted by the cell transfected with the SiRNAs was reduced with reduction of the mRNA of the La protein.Conclusion:The La protein can protect the replications of the HBV in cultured cells.

Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
Animals ; Antibodies ; analysis ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Glucosides ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; chemistry ; immunology ; Serum Albumin, Bovine ; chemistry ; immunology

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

PURPOSE: Perinatal complications associated with maternal smoking and alcohol ingestion in pregnancy have been well documented. But until now there has been no clinical data on it collected in our country. In this paper, we tried to research the association between maternal smoking and alcohol ingestion in pregnancy and the physical indices of the neonate at delivery among unmaried mothers from one charitable institution. METHODS: We enlisted 125 unmarried with a history of smoking and alcohol consumption during pregnancy who delivered a baby in National Medical Center from March, 2001 to March, 2004 as a study group. As a control, 174 unmarried mothers without a history of smoking and alcohol were enlisted. Then, we compared the physical indices - birth weight, height, and the head circumference - of the neonates from both groups. Furthermore, we investigated the effects of the period and the quantity of maternal smoking and alcohol ingestion on the physical indices of the neonates from the both groups. RESULTS: Low physical indices of neonates were found in the study group(either smoking group, or drinking group) compared with the control group. The quantity of smoking or drinking and the period of smoking or drinking have no significant effect on physical indices. CONCLUSION: We found that maternal smoking and alcohol result in the low physical index of neonates, and educational initiatives must be directed at expectant mothers to emphasize the harmful effects of smoking and alcohol ingestion in pregnancy.
Alcohol Drinking ; Birth Weight ; Drinking ; Eating ; Head ; Humans ; Illegitimacy ; Infant, Newborn* ; Mothers ; Pregnancy* ; Single Person ; Smoke* ; Smoking*

BACKGROUNDOne of the reasons for poor neuroregeneration after central nervous system injury is the presence of inhibitory factors such as Nogo. Here, we tested the inhibition of Nogo by RNA interference both in vitro and in vivo, using recombinant adenovirus-mediated transfection of short hairpin RNAs, to explore a new method of treatment for spinal cord injury.

METHODSWe designed and cloned two Nogo-specific short hairpin RNAs and an unrelated short hairpin RNA, packaged the clones into adenovirus, and amplified the recombinant virus in 293 cells. We then tested the inhibition of Nogo expression both in vitro in adenovirus-transfected oligodendrocytes and in vivo in spinal cord tissue from adenovirus-transfected spinal cord injury model rats. We tested Nogo expression at the mRNA level by reverse-transcription PCR and at the protein level by Western blotting and immunohistochemistry.

RESULTSIn vitro, the two specific Nogo short hairpin RNAs decreased Nogo mRNA expression by 51% and 49%, respectively, compared with Nogo expression in cells transfected with the unrelated control small hairpin RNA (P < 0.005). Similarly, Nogo protein expression decreased by 50% and 48%, respectively (P < 0.005). In vivo, in spinal cord injury model rats, the two specific Nogo short hairpin RNAs decreased Nogo mRNA expression by 45% and 40%, respectively, compared with Nogo expression in spinal cord injury model rats transfected with the unrelated control short hairpin RNA (P < 0.005). The Nogo protein level was similarly decreased.

CONCLUSIONSWe were successful in specifically downregulating Nogo at the mRNA and protein levels by adenovirus-mediated delivery of short hairpin RNAs, both in vitro and in vivo. This confirms the effectiveness of RNA interference for the inhibition of Nogo gene expression and the efficiency of using adenovirus for delivery. Thus gene therapy may be an effective treatment for spinal cord injury.

Adenoviridae ; genetics ; Animals ; Blotting, Western ; Humans ; Immunohistochemistry ; Myelin Proteins ; genetics ; metabolism ; Nogo Proteins ; RNA Interference ; RNA, Small Interfering ; genetics ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; therapy