Vitrification provides a rapid cooling to induce glass-like solidification inside cells and protect cell membrane and cytoskeleton system free of injury by ice crystals.The main factors that can influence the effect of vitrification are the size of ovarian tissue, the kind of cryoprotectants, the method of permeation, carrier system and so on. Ovarian tissue structure is very complex, and there is no uniform standardized protocol of vitrification yet.This paper presents also the current problems of ovarian tissue vitrification.

Objective To introduce the application and assess the outcome of ProPlan CMF software in virtual surgery of mandibular resection and reconstruction with vascularized fibular graft.Methods Nineteen patients were performed mandibular resection,and immediate reconstruction with free fibular graft by application of ProPlan CMF software between March 1,2013 and January 31,2014,including 7 cases of osteoradionecrosis,6 cases of adamantoblastoma,4 cases of carcinoma of gingiva,2 cases of intra-osseous carcinoma of mandible.There were 12 males and 7 females,with an age range of 21-73 years (median,54 years).In each case,three-dimensional spiral CT scan of mandible and fibula was obtained before operation.The CT data were imported into the ProPlan CMF software and the virtual surgical planning was performed.After that,the mandibular rapid prototyping and surgical guide plates were made according to customized design.The segmental resection of mandible,titanium plate bending,fibular molding,insetting and fixing were then carried out by using these surgical guide plates.Results Operations were undertaken successfully according to preoperative design in all nineteen patients and negative margin was detected by intraoperative frozen section.The fibula was cut,shaped and fixed accurately.The operations took 4.5-7.0 hours (mean,5.5 hours).Primary healing of incision was obtained without complications except fistula in 2 cases of osteoradionecrosis,which were resolved with local wound care after 4 weeks.19 patients were followed up between 3 months to 1 year and all patients were alive with disease free.All patients were satisfactory with their maxillofacial appearance,occlusion and function.The ranges of mouth opening was 2.0-4.0 cm.The postoperative panoramic radiographs or CT scan showed perfect situation and contour of fibular graft.Healing of the donor site occurred without significant complications.All patients started early postoperative ambulation,and walked normally after 2 or 3 months.Conclusion The application of ProPlan CMF software in virtual surgical design and guide plates creation can simplify the operation,and improve the accuracy of mandibular resection and reconstruction,especially the spatial relationship of the mandible and the fibula graft.It is worth promotion widely in clinical practice.

Objective:To analyze the inhibiting mechanism of microRNA-199a/b-3p(miR-199)on cell proliferation of triple negative breast cancer(TNBC)cells.Methods:Expression of miR199 in BT549 and MDA-MB-231 cells transfected with miR-199a/b-3p was detected by qRT-PCR.The proliferation of BT549 and MDA-MB-231 cells transfected with miR-199 were analysed by MTT as-say.Cell cycle of TNBC cells transfected with miR-199 was detected by Flow Cytometry assay.Results:MiR-199a/b-3p could suppress the proliferation of BT549 and MDA-MB-231 cells.Comparing with normal control,the proliferation rate were up to(41.02±2.34)%and(28.42 ±6.70 )%,and the cell cycle were arrest at G 1 phase.Conclusion: MiR-199a/b-3p could suppress the proliferation of TNBC,and may be a promising anti-cancer drug for TNBC.

ObjectiveTo investigate the early clinical risk factors of severe acute inhaled organic fluorine poisoning.Methods The clinical data of patients with acute poisoning of organic fluorine inhalation admitted since 2004 in Northern Jiangsu People's Hospital were retrospectively analyzed. According toDiagnostic Criteria of Occupational Acute Fluorohydrocarbon Poisoning(GBZ66-2002), all the patients were divided into three groups: mild, moderate and severe poisoning groups, the severe cases were included in the intensive group, and the others were grouped in the non-intensive group. The contents in the survey were as follows: gender, age, vital signs on admission (body temperature, pulse rate, respiratory rate, systolic blood pressure), arterial blood gas analysis record〔arterial oxygen saturation(SaO2), oxygenation index(PaO2/FiO2), lactic acid(Lac) and arterial partial pressure of carbon dioxide(PaCO2), pH value(pH)〕. Before treatment, the white blood cell(WBC) count, platelet(PLT) count, levels of alanine transaminase(ALT), creatinine(Cr), blood glucose, electrolytes(potassium, sodium, chloride, calcium), creatine kinase isoenzyme(CK-MB), etc. were examined and recorded. All the patients were immediately arranged for bedside chest X-ray examination, and the chest X-ray lung injury scores were recorded. By univariate and multivariate logistic regression analyses, the receiver operating characteristic curve(ROC curve) was drawn to evaluate the diagnostic value of the clinical risk factors.Results Sixty-two cases consisting with the standard criteria of enrollment were collected in the study, 36 cases being in intensive group and 26 cases in non-intensive group. The univariate analysis showed that the differences in pulse rate, respiratory rate, PaO2/FiO2, WBC, SaO2, Lac, pH, and lung injury score were statistically significant(P<0.05 orP<0.01). Logistic multiple regression analysis showed that PaO2/FiO2, WBC, Lac and chest X-ray lung injury score were the four indexes for predicting the independent risk factors of severe acute inhaled organic fluorine poisoning. The area under ROC curve(AUC) of PaO2/FiO2 was the highest(0.884), 95% confidence interval(95%CI) was 0.784 - 0.984, the critical value was 96.5 mmHg(1 mmHg=0.133 kPa), with the sensitivity of 75.6%, specificity of 95.2%, positive predictive value(PPV) of 92.3% and the negative predictive value(NPV) of 71.4%, in sequence, the rest were WBC(AUC 0.846, 95%CI 0.728 - 0.965, the criticalvalue 12.15×109/L), Lac(AUC 0.800, 95%CI 0.662 - 0.938, the critical value 4.2 mmol/L), chest X-ray lung injury score(AUC 0.795, 95%CI 0.652 - 0.938, the critical value 2.50), the sensitivity of the above three items was 90.2%, 83.6%, 88.5%, specificity was 90.2%, 83.6%, 88.5%, the PPV was 86.7%, 82.4%, 85.8% and NPV was 72.0%, 73.9%, 69.2% respectively.ConclusionThe blood WBC count, Lac, PaO2/FiO2 and chest X-ray lung injury score can be used as the early clinical risk factors of severe acute inhaled organic fluorine poisoning.

Objective To discuss the effect of in vitro fertilization ( IVF) and mouse sperm cryopreservation , to establish a simple and economic frozen system for the genetically engineering mice preservation .Methods Sperm from genetically engineering mice were cryopreserved , IVF was performed using post-thawed sperm, then embryo transfer, to compare the effects of cryopreservation medium、age of male mice and sperm preincubation medium .Results Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , the fertility rates were from 82.49%to 91.43%, when HTF was used thawed-sperm preincubation in IVF , the fertility rates were from14.46%to 27.38%, there was a signification difference between PM and HTF sperm preincubation medium;10 to 35 weeks male genetically engineering mice sperm were succeed cryopreservation , and positive mice were procreated after 2-cell embryos were transferred;R18S3、CPM and CPA was used to freeze sperm , the fertility rates were 75.85%、88.89%to 94.27%, positive mice were procreated after 2-cell embryos were transferred;2-cell embryos after IVF were freezed , then thawed and positive mice were procreated after 2-cell embryos were transferred .Conclusion Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , genetically engineering mice sperm were succeed cryopreservation .

Objecfive To investigate possibihty of monitoring myocardial isehemia by eontinuous EASI 12-lead ST-segment trend analysis during orthopaedic surgery and find its rules.Methods Three hundred and eleven consecutive patients who underwent orthopaedic surgery were monitored bv continuous EASI 12-lead ST-segment analysis during surgery.Results Among 311 patients 81 patients(26.0%) had myocardial ischemic events.The ischemie incidence of general anesthetics WaS signifieantly higher than local anesthetics(the spinal or epidural anesthetics and nerve-block anesthetics)(38.1%vs 23.0%.P＝ 0.01 5).During ischemie events the patients whose heart rate exceeded 90 beat per minute was 71.6% (58/81).Among the 177 isehemie events,the duration ofless than 5 minutes was 79.7%(141/177).ST-seg- ment deviation ofless than 0.15 mV was 71.2%(126/177).Conclusions During orthopaedic surgery EASt 12-lead electrocardiogram is the optimal approach for monitoring myocardial isehemia.The ischemic incidence of general anesthetics is significantly higher than local anesthetics.The myocardial ischemia events correlates with fast heart rate.The intraoperative myocardial ischemia events are more often transient.In most of these ischemic events ST-segment deviation is not very serious.

Objective Cell-free DNA (cfDNA) was shown to be a prognostic marker for diverse pathological states in the Intense Care Unit, but little is known of the role of cfDNA in HBV-related acute-on-chronic liver failure (ACLF). We hypothesize that cfDNA can also be a promising prognostic as well as a diagnostic marker in patients with HBV-related ACLF. Methods Thirty-eight patients with HBV-related ACLF admitted in the Intense Care Unit were enrolled in the study. The patients were divided, according to the improvement of liver function at discharge, into favorable prognosis group (group 1, n=17) and poor prognosis group (group 2, n=19). Plasma samples were collected from each patient at hospitalization and at discharge to measure cfDNA by real-time quantitative PCR. MELD score was calculated at the same time points. Results The average level of cfDNA of group 1 was lower than that of group 2 both at the time of hospitalization (P=0.044) and at discharge (P<0.001). There was no difference in MELD score between the two groups at hospitalization. Significant correlations were found of cfDNA levels with the MELD score, TBIL, CRE and INR both at hospitalization (γ=0.662, P<0.001;γ=0.356, P=0.033;γ=0.360, P=0.031;γ=0.570, P<0.001, respectively) and at discharge (γ=0.854, P<0.001;γ=0.821, P<0.001;γ=0.650, P<0.001;γ=0.638, P<0.001, respectively). The ROC curve showed that cfDNA level at discharge was optimal in diagnosing ACLF with an area under curve (AUC) value of 0.96, followed by?cfDNA (AUC value of 0.923) and cfDNA level at hospitalization (AUC value of 0.667). The MELD scores had an AUC value of only 0.545 at the time of hospitalization. Conclusion cfDNA may serve as a promising prognostic and diagnostic marker for predicting in-hospital prognosis of HBV-related ACLF within 2 to 8 weeks.

Objective Cell-free DNA (cfDNA) was shown to be a prognostic marker for diverse pathological states in the Intense Care Unit, but little is known of the role of cfDNA in HBV-related acute-on-chronic liver failure (ACLF). We hypothesize that cfDNA can also be a promising prognostic as well as a diagnostic marker in patients with HBV-related ACLF. Methods Thirty-eight patients with HBV-related ACLF admitted in the Intense Care Unit were enrolled in the study. The patients were divided, according to the improvement of liver function at discharge, into favorable prognosis group (group 1, n=17) and poor prognosis group (group 2, n=19). Plasma samples were collected from each patient at hospitalization and at discharge to measure cfDNA by real-time quantitative PCR. MELD score was calculated at the same time points. Results The average level of cfDNA of group 1 was lower than that of group 2 both at the time of hospitalization (P=0.044) and at discharge (P<0.001). There was no difference in MELD score between the two groups at hospitalization. Significant correlations were found of cfDNA levels with the MELD score, TBIL, CRE and INR both at hospitalization (γ=0.662, P<0.001;γ=0.356, P=0.033;γ=0.360, P=0.031;γ=0.570, P<0.001, respectively) and at discharge (γ=0.854, P<0.001;γ=0.821, P<0.001;γ=0.650, P<0.001;γ=0.638, P<0.001, respectively). The ROC curve showed that cfDNA level at discharge was optimal in diagnosing ACLF with an area under curve (AUC) value of 0.96, followed by?cfDNA (AUC value of 0.923) and cfDNA level at hospitalization (AUC value of 0.667). The MELD scores had an AUC value of only 0.545 at the time of hospitalization. Conclusion cfDNA may serve as a promising prognostic and diagnostic marker for predicting in-hospital prognosis of HBV-related ACLF within 2 to 8 weeks.

OBJECTIVETo investigate the effects of Bushen Huoxue Fang on the proliferation of rat cardiac fibroblasts and collagen production in the cells.

METHODSRat cardiac fibroblasts were isolated and cultured in DMEM containing 10% (group A) or 20% (group B) or no (group C) serum from rats treated with Bushen Huoxue Fang, with cells cultured in DMEM containing 10% FBS as the control (group D). After 72 h of cell culture, the proliferation of the fibroblasts was detected using CCK-8 kit, and collagen mRNA and protein expressions were examined using RT-PCR and Western blotting, respectively.

RESULTSCompared with that in groups C and D, the cell proliferation decreased significantly in groups A and B, and especially in the latter (P<0.05). RT-PCR demonstrated significant reductions of the mRNAs of type 1 and 3 collagens in groups A and B (P<0.05), and their protein levels were also significantly lowered (P<0.05).

CONCLUSIONBushen Huoxue Fang can effectively inhibit the proliferation of rat cardiac fibroblasts and reduced collagen type 1 and 3 productions in the cells in vitro.

Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Fibrosis ; prevention & control ; Myocardium ; cytology ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVECell-free DNA (cfDNA) was shown to be a prognostic marker for diverse pathological states in the Intense Care Unit, but little is known of the role of cfDNA in HBV-related acute-on-chronic liver failure (ACLF). We hypothesize that cfDNA can also be a promising prognostic as well as a diagnostic marker in patients with HBV-related ACLF.

METHODSThirty-eight patients with HBV-related ACLF admitted in the Intense Care Unit were enrolled in the study. The patients were divided, according to the improvement of liver function at discharge, into favorable prognosis group (group 1, n=17) and poor prognosis group (group 2, n=19). Plasma samples were collected from each patient at hospitalization and at discharge to measure cfDNA by real-time quantitative PCR. MELD score was calculated at the same time points.

RESULTSThe average level of cfDNA of group 1 was lower than that of group 2 both at the time of hospitalization (P=0.044) and at discharge (P<0.001). There was no difference in MELD score between the two groups at hospitalization. Significant correlations were found of cfDNA levels with the MELD score, TBIL, CRE and INR both at hospitalization (γ=0.662, P<0.001; γ=0.356, P=0.033; γ=0.360, P=0.031; γ=0.570, P<0.001, respectively) and at discharge (γ=0.854, P<0.001; γ=0.821, P<0.001; γ=0.650, P<0.001; γ=0.638, P<0.001, respectively). The ROC curve showed that cfDNA level at discharge was optimal in diagnosing ACLF with an area under curve (AUC) value of 0.96, followed by δcfDNA (AUC value of 0.923) and cfDNA level at hospitalization (AUC value of 0.667). The MELD scores had an AUC value of only 0.545 at the time of hospitalization.

CONCLUSIONcfDNA may serve as a promising prognostic and diagnostic marker for predicting in-hospital prognosis of HBV-related ACLF within 2 to 8 weeks.

Acute-On-Chronic Liver Failure ; diagnosis ; virology ; Adult ; DNA, Viral ; blood ; End Stage Liver Disease ; diagnosis ; Female ; Hepatitis B ; complications ; Hepatitis B virus ; genetics ; Humans ; Male ; Middle Aged ; Pilot Projects ; Plasma ; chemistry ; Prognosis ; Severity of Illness Index