BACKGROUND:As the bone and marrow tissue have very special structure, it is difficult to simultaneously display the bone with tough hard tissue and bone marrow tissues containing various immature hematopoietic cel s in the conventional process of pathological section preparation. OBJECTIVE:To choose the best decalcifying solution that cannot only completely remove the calcium in the bone tissue but also protect the structure of bone marrow tissues and cel s from damage. METHODS:Bone marrow tissues from the long bone of dogs were randomly divided into four groups. Under the same conditions, the bone marrow tissues were decalcified with 14%formaldehyde saline solution of nitric acid (group A), 14%nitric acid solution (group B), 20%A saline solution of hydrochloric acid formaldehyde (group C) and 20%A hydrochloric acid aqueous solution (group D). Decalcified time was recorded, fol owed by routine dehydration, section, hematoxylin-eosin staining and microscopic observation. Pathological section quality and hematoxylin-eosin staining were compared among the four groups. RESULTS AND CONCLUSION:Group A had the best sections and hematoxylin-eosin staining, strongest decalcified ability, shortest decalcified time and minimum damage to the bone marrow. Group B had the worst results of section and hematoxylin-eosin staining, in which, the bone tissues were loose and became yel ow and the bone marrow tissue were damaged greatly, and the decalcified effect was worse. Group C was worse than group A in decalcified ability, damage degree, section quality and hematoxylin-eosin staining results. Group D also had a better result of section and hematoxylin-eosin staining as wel as exhibited uniform decalcification effect and less damage to the bone marrow, which was ranked between group B and group C. Al the four kinds of decalcifying solutions have a good decalcification ability, but the section quality and hematoxylin-eosin staining results rank as fol ows:Group A>Group C>Group D>Group B. Taken together, 14%formaldehyde saline solution of nitric acid is ideal for the clinical preparation of pathological sections.

Objective: Our purpose was to study the relationship between the Fas antigen expression and neuron apoptosis after cerebral ischemia. Methods: We detected the Fas antigen expression and neuron apoptosis dynamically in the animal models with cerebral ischemia by immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. Results: Fas antigen was positive after 30 minutes of ischemia and reached peak at 60 minutes. At the 24th hour, it began to decrease, and negative on the 3th day. While the positive cell for TUNEL method appeared after 60 minutes of ischemia, reached peak on 3 day, and decreased on 7 day. Conclusion: Neuron apoptosis after cerebral ischemia is closely related to the over-expression of Fas antigen.

The in-vitro fertilization and embryo transfer technique has been widely applied in human insemination. The rate of successful insemination is gradual y rising, and the in-vitro fertilization directly determine the insemination outcome.
OBJECTIVE:To evaluate the difference between the two common using insemination methods, microdrop and open, in in-vitro fertilization and embryo development.
METHODS:A randomized study was conducted to compare microdrop and open insemination methods among non-male factor patients undergoing in-vitro fertilization and embryo transfer. A total of 1 175 cases were enrol ed in the research. There were 573 cases in the microdrop group, and 602 cases in open insemination group. The fertilization rate and embryo development in the two groups were compared.
RESULTS AND CONCLUSION:The fertilization failure rate [total fertilization failure rate+low fertilization rate (<25%oocytes fertilized)] in the microdrop insemination group was higher than in the open insemination group (11.9%, 3.3%, P<0.001), while the good quality embryo rate and pregnancy rate did not differ significantly between the two groups (al P>0.05). The open insemination method is a simple insemination method with a lower fertilization failure rate. As the fertilization is a highly complicated process involving many extrinsic and intrinsic factors, further study is needed to confirm the effects of the two insemination methods on in-vitro fertilization outcome.

Objective To develop a HPLC method for determining the contents of lobetyolin and gallic acid in Eighteen Flavors Dangshen Pill(EFDSP) produced by different factories.Methods The HPLC analysis was performed on a VP-DOS C18 column (4.6 mm×150 mm,5 μm).The mobile phase was acetonitrile and 0.4% glacial acetic acid(21∶79) in the determination of lobetyolin content.The detection wavelength was 267 nm and the flow velocity was 1 mL/min.the column temperature was25 ℃ and the sample size was10 μL.The mobile phase was methanol and 0.4% glacial acetic acid(1∶99) in the determination of gallic acid content.The detection wavelength was 280 nm.The column temperature was 25 ℃ and the sample size was 10μL.Results The contents of lobetyolin and gallic acid in EFDSP were 1.0835mg·g-1and 15.334 0 mg/g for Qinghai Gela Dandong Tibetan Pharmaceutical Factory;0.628 9 mg/g and 15.159 5 mg/g for Changdu Tibet Medicine Factory;0.306 5 mg/g and 8.762 7 mg/g for Tibetan Hospital of Tibet.Conclusion This method has the advantages of good reproducibility,good accuracy,simple and fast operation.The contents of lobetyolin and gallic acid in EFDSP produced by different manufacturers are significantly different.The gallic acid content has greater difference.It provides the reference for quality control of EFDSP

Objective To investigate the role of C-Jun N-terminal kinase (JNK) in epithelial mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells(RPMCs). Methods RPMCs were harvested from the peritoneum of male Sprague-Dawley rats, then cultured in DMEM/F12 medium with 15% (V/V) FBS. After stimulation with TGF-β1, the expression of a-smooth muscle actin (α-SMA), E-cadherin and collagen I were detected in RPMCs. In some groups, the ceils were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGF-β1. The protein expression of phosphorylated JNK was detected by Western blotting. The mRNA and protein expression ofα-SMA, E-cadherin and collagen I were examined with RT-PCR and Western blotting, respectively.The intracellular distribution and expression of α-SMA was determined by indirect immunofluorescence. Results TGF-β1 could significantly increase the expression of α-SMA and collagen I, and decrease the expression of E-cadherin in RPMCs. TGF-α1 could stimulate the expression of phosphorylated JNK at 5 minutes with the peak at 10 minutes (P<0.01). The addition of SP600125 effectively inhibited TGF-β1-induced high expression of α-SMA and collagen I (P<0.05), and prevented TGF-β1-induced down-regulation of E-cadherin expression in RPMCs (P<0.05). The indirect immunofluorescence showed that the expression of intracellular α-SMA in RPMCs stimulated by TGF-β1 for 48 h increased significantly, which could be inhibited by SP600125. Conclusions JNK regulates epithelial mesenchymal transition induced by TGF-β1 in rat peritoneal mesothelial cells. JNK inhibitor may be used as a novel therapeutic agent for peritoneal fibrosis.

Objective To analyze the common causes leading to lung infection following renal transplantation and provide targeted preventive measures to reduce the incidence of lung infection.Methods The clinical data of 561 recipients who underwent renal transplantation from January 2006 to February 2011 were retrospectively analyzed.The recipients were divided into two groups:group Ⅰ,from January 2006 to December 2009 (n =416) ; group Ⅱ,from January 2010 to February 2011 (n =145).The causes possibly leading to lung infection which took place 3 days before the appearance of the clinical symptoms were offered by the patients who suffered lung infection of group Ⅰ.And then the causes were summarized and analyzed to formulate the specific and comprehensive measures to prevent the infection.Finally the measures were applied to recipients in group Ⅱ from January 2010.After applying the measures for 14 months,the incidence of lung infection in group Ⅱ was counted and compared with that in group Ⅰ to see the preventive effect.Results There were 58 cases of lung infection in group Ⅰ (58/416,13.9％) and 12 cases in group Ⅱ (12/145,8.3％). There was significant difference in the incidence of lung infection between two groups (x2 =4.0361,P＜0.05).All of the recipients with lung infection were hospitalized in six months after the transplantation.The causes leading to lung infection of 58 cases in group Ⅰ were as follows:6 cases due to being excessively tired,3 cases due to guest visiting,12 cases due to abrupt change of weather,9 cases due to exposure to public place,8 cases due to returning to hospital,6 cases due to close contact with children,5 cases due to close contact with animals,and the other 9 cases without specific causes found.Conclusion The incidence of lung infection following renal transplantation can be notably reduced by the application of targeted and concrete health propaganda education and preventive measures based on analysis on the specific causes of infection.

?AIM:To compare the coherence and difference on the fundus examination made with two kinds of optical coherence tomography ( OCT): Angio-OCT and fourier domain-optical coherence tomography ( FD-OCT) .?METHODS:Using Angio-OCT and FD-OCT to measure the retinal nerve fiber layer ( RNFL ) thickness, optic parameters, and ganglion cell complexes ( GCC ) thickness from 20 subjects respectively.The coherence was tested with Pearson's correlation coefficient, the difference was tested with paired Student t testing.?RESULTS:The total correlation of the RNFL thickness, optic parameters, GCC thickness made with two kinds of OCT was between 0.7-0.8;the RNFL thickness, optic disk area etc.made with the Angio-OCT were lower than those made with FD-OCT except for the GCC thickness.?CONCLUSION: The results made with two kinds of OCT from the same subject has certain coherence, but cannot be compared directly.

Objective To study the safety and efficacy of continuous intravenous infusion of fentanyl in laser photocoagulation of retinopathy of prematurity (ROP).Methods From March,2014 to January,2015,ROP infants hospitalized for laser photocoagulation in Guangdong Women and Children hospital were randomly (using envelope method)assigned into the fentanyl groupand the control group.In the fentanyl group,the patients were given fentanyl combined with topical anesthesia,while onlytropical anesthesiain the control group.Premature infant pain profile (PIPP)scores,heart rate,mean artery pressure and complications within 3 days after operation were recorded. The concentration of epinephrine, norepinephrine and cortisol in the blood were measured before and after the operation.Student′s t test,non-parameter rank and chi-square test were used to compare the differences between the two groups.Results A total of 82 infants were enrolled in the study,42 in the fentanyl group and 40 in the control group.In the fentanyl group,11 .9% infants had maximum PIPP score ≥6 and 70.0% in the control group,the difference was statistically significant (P <0.05).In the fentanyl group,the PIPP score at the beginning of operation, the maximum PIPP score during operation and the PIPP score at the end of operation were 2.0,3.0and 1 .5, respectively.In the control group,these scoreswere 8.0,8.0and 8.0 respectively.The differences were statistically significant (P <0.05 ).No significant differences existed between the concentration of epinephrine,norepinephrine and cortisol before and after operationin the fentanyl group.However,these concentrations were elevated after the operation than before the operation in the control group (P <0.05).The incidence of complications within three days after operation was 19.0% in the fentanyl group and 40.0% in the control group,and the difference was statistically significant (P <0.05 ).Conclusion Comparing with topical anesthesia,fentanyl combined with topical anesthesia has lower pain scores,less stress responses and fewer complications during ROP laser photocoagulation.Fentanyl combined with topical anesthesia is a safe and effective analgesic method during ROP laser photocoagulation.

Objective To preliminarily understand the genotype characteristics of Toxoplasma gondii in blood of HIV?posi?tive persons in Lincang City,Yunnan Province. Method Two segments of SAG2 gene of T. gondii from blood samples of HIV?positive persons in Lincang City were extracted and amplified by using the nested PCR method and the genotype was identified and compared with the standard strain(Type I)of Toxoplasma gondii. Results Thirty?five SAG2 genes(241 bp)and 35 SAG2 genes(221 bp)of T. gondii were amplified from 170 blood samples of the HIV?positive people,and 4 of each case were selected and digested with enzyme,then 2 aim gene fragments of each case were chosen and compared with the standard strain (Type I)of T. gondii. The digestion of SAG2 gene(241 bp)showed the genotype of the blood samples was Type I or Type II, and the digestion of SAG2 gene(221 bp)confirmed that the genotype was Type I. Conclusion It is preliminarily confirmed that the genotype of T. gondii in blood of HIV?positive persons in Lincang City,Yunnan Province is Type I.

ObjectivesTo investigate the clinical value of the melting curve analysis-based PCR assay for the clinical genetic diagnosis and prenatal diagnosis of β-thalassemia.Methods A total of 451 peripheral blood samples,including 372 cases with β-thalassemia phenotypes and 79 cases without β-thalassemia phenotypes,were collected by Liuzhou Municipal Maternity and Child Healthcare Hospital between January 2011 and August 2011.Moreover,another 84 cases,including 16 fetal villi samples (10 - 13 weeks),64 amniotic fluid samples (16 -24 weeks ) and 4 umbilical cord blood samples (above 17 weeks),whose parents were β-thalassemia carriers,were also collected for this assay between June 2011 and September 2011.A double-blind test was done to compare the detection reliability of the melting curve analysis-based assay (24 β-thalassemia mutations can be detected) with PCR-RDB probe assay (17 β-thalassemia mutations can be detected ) and DNA sequencing using these samples.The wildtype,mutant and total concordance rates of the genotyping results were calculated separately among the melting curve analysis based assay,PCR-RDB probe assay and DNA sequencing.Results Among the 451 peripheral blood samples,thirteen mutations and nineteen genotypes were obtained by using melting curve analysis-based assay.447 samples had the same detection results and 4 samples had different detection results by comparing melting curve analysis-based assay with PCR-RDB probe assay,thus,the concordance rate of the sample detection result was 99.1％ (447/451),and the concordance rate of the allele detection result was 99.6％ (898/902).DNA sequencing results of the 4 samples showed that 3 samples had the same genotyping result with melting curve analysis-based assay,and 1 sample had the same genotyping result with PCR-RDB probe assay.A rare β-globin mutation which was not included by melting curve analysis-based assay was not detected.Thus,the genotypes of 450 samples were detected accurately by melting curve analysis-based assay,and the concordance rate of the sample detection between the melting curve assay and DNA sequencing assay was 99.8％ (450/451).Among 84 fetal villi,amniotic fluid,and umbilical cord blood samples,8 mutation types and 18 genotypes were obtained by using melting curve analysis-based assay.All the samples have the same detection results by comparing melting curve analysis-based assay with PCR-RDB probe assay and DNA sequencing,so the concordance rate of the genotyping results was 100％ among the melting curve analysis-based assay,PCR-RDB probe assay and DNA sequencing.Conclusions The melting curve analysis-based PCR assay can detect multiple unknown samples simultaneously,and detect multiple mutations accurately.It is very useful for the genetic diagnosis and prenatal diagnosis of β-thalassemia.