BACKGROUND:The successful establishment of human embryonic stem cel lines in vitro is of great significance to human embryonic development mechanism and developmental biology, cel and tissue transplantation in the treatment of certain diseases. OBJECTIVE:To summarize the progress of in vitro culture of embryos and establishment of embryonic stem cel lines, to explore the influential factors for in vitro culture of embryos, and the methods of culturing human discarded embryos, isolating inner cel mass and establishing embryonic stem cel lines, as wel as the establishing conditions for embryonic stem cel lines. METHODS:With the key words of“embryo, embryonic stem cel s, coculture, sequential culture”, the first author searched CNKI and SCI databases for literatures concerning in vitro culture and transplantation of embryos and establishment of embryonic stem cel lines published from 2000 to 2014. Systematic evaluation was conducted. Final y, 58 literatures were retained for result analysis. RESULTS AND CONCLUSION:The culturing condition for embryos in vitro is the key factor affecting embryo transfer outcomes, including culture medium component and culture system. In previous studies, the component and application of culture medium have changed greatly, and the culture system has altered from single culture to coculture and sequential culture. Ethical issues and embryonic origin restrictions restrict the establishment of human embryonic stem cel lines. Clinical y discarded low-quality embryos can be used as one of the material sources to establish human embryonic stem cel lines, which can effectively lessen the problem of embryo shortage during the establishment of human embryonic stem cel lines and reduce ethical disputes.

BACKGROUND: Early mammalian embryonic development and implantation is a very complex process, which is regulated by a variety of cytokines. Moreover, epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor and vascular endothelial growth factor play an important role in this regulation process.
OBJECTIVE: To introduce cytokines related to early embryonic development, including types, structural features and molecular mechanisms, and meanwhile to review the effects of epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor and vascular endothelial growth factor in the early embryonic development.
METHODS:A literature retrieval was performed to search relevant articles published from 1989 to 2015 using the keywords of “embryos development, epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor, vascular endothelial growth factor”. Active effects of these five kinds of cytokines in overcoming the mammalian embryonic development block were reviewed and summarized to provide theoretical and experimental basis for further optimization of the embryonic culture system in vitro.
RESULTS AND CONCLUSION: Cel growth factors involved in embryonic development is the focus of the study. Insulin-like growth factor is capable of promoting the embryonic growth; epidermal growth factor exhibits multiform functions in the embryonic development; leukemia inhibitory factor directly affects oocyte cel development, and progesterone increases the expression of leukemia inhibitory factor to provide a good cytokine microenvironment for embryonic development; vascular endothelial growth factor promotesin vitro oocyte maturation, and plays a very important role in various stages of embryonic development. To clarify these regulatory patterns of cytokines cannot only contribute toin vitro embryonic culture, but also reduce embryonic apoptosis, which is of great significance for promoting embryonic development.

objective: To observe the distribution of pegylated liposmal doxorubicin(PLD) in animal model of tongue cancer. Methods: Tongue cancer model was established in 40 golden hamster, PLD or free doxorubicin at the dose of 9 mg/kg was injected perifocally into each of 20 mice, the concentration of the drugs in lypmph node and blood was measured with HPLC. Results: After injection of free ADM peak concerntration in blood was acheived in 3 h and in lymph nodes in 1 h, while after injection of PLD, that was in 16 and 6 h respectively. The concertration of PLD was heigher than that of ADM in both blood and lymph node from 16 or 6 h till 216 h after injetion.Conclusion: PLD can be regarded as valuable drug delivery system in the treatment of oral cancer.

Objective To evaluate the diagnostic value of magnetic resonance imaging (MRI) on fetal ventriculomegaly identified through prenatal ultrasonography and the outcomes of these newborns were followed up. Methods From March 2006 to July 2008, MRI was performed on 135 pregnant women whose fetuses diagnosed as fetal ventriculomegaly at an average of 32 gestational weeks in Shengjing Hospital Affiliated to China Medical University. Mild ventriculomegaly was defined when the width of unilateral or bilaeral fetal cerebral ventricle triangle was 10-15 mm, moderate ventriculomegaly 16-20 mm and severe ventriculomegaly >20 mm. We introduced the Denver developmental screening test(DDST) to follow-up the mild ventriculomegaly and normal babies, confirmed by MRI, at 6-12 months after birth and a case-control study was conducted. The intelligence and growth of these infants were analyzed. Results (1) Diagnostic rate of fetal ventriculomegaly through MRI: Among the 135 gravidas, 60 (44.4%) showed isolated ventriculomegaly, 5 (3.7%) complicated with ventricular hemorrhage; 12 (8.9%) complicated with agenesis of corpus callosum (ACC) and 2 (1.5%) complicated with cerebellar hypoplasia, while 56 (41.5%) were normal. Seventy-nine cases had fetal ventriculomegaly on MRI and 15.2% (n=12) of them complicated with ACC. (2) Degree of fetal ventriculomegaly on MRI: Among the 60 isolated ventriculomegaly cases, 55 (91.7%) were mild and 5 (8.3%) moderate ones. Among the 5 cases complicated with ventricular hemorrhage, one was mild ventriculomegaly, and 4 moderate or severe cases. Among the 12 cases with ACC, 8(66.7%) were moderate ventriculomegaly and 4 (33.3%) severe cases. The 2 cases with cerebellar hypoplasia were both moderate ventriculomegaly fetuses. (3) Follow-up at 6-12 months after birth : thirty out (case group) of the 55 isolated ventriculomegaly cases, 38 out of the 56 normal babies and 42 babies with normal MRI results were followed up, and the later 80 cases were taken as control. Four infants (13.3%) in the case group and 10 (12.5%) in the control group showed abnormal or suspected results in DDST (P>0.05), the rest babies were all normal. (4) Clinical outcomes of the 79 ventriculomegaly fetuses diagnosed by MRI: thirty mild ventriculomegaly babies and 5 moderate ones were born at term and showed normal at follow ups. However, 7 gravidas were not compliant, 6 pregnancies were terminated, and 12 were last. Three of the 12 cases with ACC continued the pregnancy, and postnatal MRI of the babies showed the same with the prenatal MRI, 8 pregnancies were induced and one was lost. All of the 5 fetuses with ventricufar hemorrhage were induced and the prenatal diagnosis was confirmed by autopsy. One of the 2 fetuses with cerebellar hypoplasia was term delivered and diagnosed as cerebral palsy at the age of 6 months, and the other one was induced. Conclusions MRI is an indispensable complementary diagnostic method for fetal ventriculomegaly diagnosed through ultrasound. The development of intelligence and growth of babies born with mild isolated ventriculomegaly is the same as normal ones.

Objective To evaluate the diagnostic value of MRI in the cases suspected of ventriculomegaly by prenatal uhrasonography.Methods 104 patients of suspected fetal ventriculomegaly (VM) diagnosed by uhrasonography were included from the Shengjing Hospital,China Medical University from March 2006 to October 2007.All cases were divided into 4 groups based on the standard of Gaglioti:10-12 mm(66 cases),13-15 mm(22 cases),16-20 mm(14 cases),and 21-25 mm(2 cases);they included 75 eases of single intracerebroventricular expansion and 29 eases of double intracerebroventricular expansion.All of them were subjected to MRI scan within 48 h of uhrasonographic examination to determine the prenatal diagnosis by MRI pregnancy outcomes.Results Among the 26 072 cases who received prenatal uhrasonography,104 cases (0.39%) were VM. (1) MRI detected 3 cases (5%) in 10-12 mm group:one ease of cerebellar hypoplasia,vascular malformation,chest and abdominal anomalies each; 5 cases(23%)in 13 -15 mm group:one case of agenesis of corpus callosum (ACC),cerebral hemorrhage,cerebral hemorrhage with cerebral meningocele,cerebral meningocele,intracranial mass meningocele each; 6 cases(43% )in 16 -20 mm group:4 cases of ACC,one case of intraventricular hemorrhage and ACC combined with ventricular hemorrhage each; 2 cases in 21-25 mm group:one case of ACC and intraventricular hemorrhage each.(2) MRI detected 4 eases(5%) among 75 unilateral VM cases and 12 eases (41%) among 29 bilateral VM eases.The differences were significant (P＜0.01).MRI diagnosis rate was 15.38% (16 cases).Follow-up of the outcomes of the pregnancy showed induction of 0labor in 15 cases (14%) all of which were the same as MRI results on autopsy,full-term delivery of 88 cases,of which all the neonates were healthy.Conclusions When the expansion width is above 16 mm or bilateral VM is suspected by ultrasonography,we suggest MRI examination to determine fetal central nervous system disease.

Objective To investigate the multi-resistance to antibiotics of clinical isolated klebsiella pneumoniae.Methods The resistance to antibiotics of clinical isolated klebsiella pneumoniae were monitored.The discconfirmatory test was used to detect extended-spectrum β-lactamases(ESBLs) and cefoxitin three-dimension was used to detect AmpC β-lactamases.Results Among the isolates there were 53 strains of ESBLs-producing bacteria (49.5% ), 30 strains of AmpC-producing bacteria(28.0%), 24 strains of ESBLs + AmpC-producing bacteria (22.46%).They were high resistance to aminoglycosides,quinolones and cephalosporins.Conclusion The multi-resistance to antibiotics of clinical isolated klebsiella pneumoniae were widespread.It is important to control nosocomial infection to strengthen the detection of the epidemiology of ESBLs and AmpC β-lactamases in clinical isolates.

Objective To investigate effects of hypertensive agents on regional cerebral blood flow (rCBF)and prognosis in patients with hypertensive intracerebral hemorrhage (HICH). Method Thirty patients withFILCH were divided into group A,B and C according to mean arterial pressure(MAP) with various ranges of <10%, 10 ～ 20%, and > 20%, respectively. Patients of three groups were checked with single photon emissioncomputed tomography (SPECT),and computerized tomography of head successively, 24 hours, 3 ～ 5 days, and 12～ 15 days after admission for the assessment of therapeutic effects. The rtes of the decrease in rCBF compared be-twecn groups were analyzed with t test. Neurologic function defect scale, activities of living scale and number of re-bleeding compared between two groups were analyzed by X2 test, respectively. Results There were statisticallysignificant differences in the rate of the decrease in rCBF. The neurologic function defect scale and activities of liv-ing scale at 24 hours, 3 ～ 5 days, and 12 ～ 15 days after admission compared between group C and group A, andbetween group C and group B showed significant differences ( P < 0.05). There were statistically significant differ-enees in rebleeding during acute phase of HICH between group A and group B or group C (P < 0.05).Conclusions If the systolic blood pressure is above 185mmHg or diastolic blood pressure over 95mmHg,it is ra-tional and safe to lower the MAP by about 15% with hypotensive gents so as to minimize the impact on regionalcerebral blood flow.

Objective The aim of this study was to observe the growth difference and expression of cytochrome C of skin hair follicles in neonatal mice .Methods The morphology of different skin hair follicles of neonatal mice ( postnatal day 1-9)were observed by HE staining histology and cytochrome C was detected by immunohistochemistry .Results The skin hair follicles in different parts of neonatal mice showed differences not only in morphology but also in developmental pe -riods.Hair follicle growth in the back and tail skin had a nonlinear and growing period .After the nonlinear and growing pe-riod they began to grow rapidly .The tail development was slightly slower than that on the back .The hair follicles of vibris-sae were very special , and started to develop without a stable period .Conclusions The results of morphological observa-tion and cytochrome C immunohistochemistry demonstrate that differences exist in the hair follicle morphology and develop -mental times in the skin of different parts of the body in neonatal mice .

Objective To explore the significance of quality control after clinical laboratory analysis .Methods A total of 450 pieces of unqualified testing reports were collected from the Department of Clinical Laboratory from January 2012 to June 2014 and reasons causing unqualified testing reports were analyzed .Results In all 450 pieces of unqualified testing reports ,testing results of 169 pieces were inconsistent with results of clinical diagnosis ,accounted for 37 .6% ;149 pieces with missing or indirect inspection i‐tems ,accounted for 33 .1% ;62 pieces did not indicate staff or department sending specimens ,accounted for 13 .8% ;results of 36 pieces reached the critical value but without re‐inspection or did not indicate the re‐inspection ,accounted for 8 .0% ;18 pieces did not clarify specimens with lipid turbidity or jaundice and so on ,accounted for 4 .0% ;16 pieces marked with wrong sample types ,accoun‐ted for 3 .6% .Conclusion It is necessary to conduct quality control after clinical laboratory analysis before delivering report ,stand‐ardize operating procedures ,check every report seriously ,make clear responsibility and improve awareness of responsibility ,in order to provide a qualified testing report for clinical practice .

Objective:To observe and compare the therapeutic effect of metoprolol by routine increasing dose method and rapid titration method on acute myocardial infarction (AMI).Methods:A total of 60 inpatients,who were di-agnosed with AMI within 24h and without contraindications for metoprolol,were randomly divided into two groups:routine therapy group (received metoprolol using routine methods,the dose was added in seven days)and rapid ti-tration group (metoprolol was added in three days using titration).The dosage maintained with 190 mg/d after both groups reaching the target dose of 190mg/d;then therapeutic effects were observed in both groups.Results: ①There were no re-myocardial infarction,rehospitalization caused by heart failure and sudden death etc.in both groups;② Patients received echocardiography in outpatients after three months.Compared with routine increasing dose group,there was significant reduction in left ventricular end-diastolic diameter [LVEDd,(55.00±7.56)mm vs.(50.00± 5.81)mm]and significant rise in left ventricular ejection fraction [LVEF,(49.13 ± 10.18)% vs. (57.84±10.34)%]in rapid titration group,P <0.01 both.Conclusion:Rapid titration method could make the pa-tients rapidly reach the targeted dose of metoprolol and inhibit renin release earlier,block the renin-angiotensin sys-tem,and improve myocardial remodeling and cardiac function.