OBJECTIVETo evaluate the functional outcomes of modified limited "L" incision beside the Achilles tendon with distraction bone block arthrodesis in treatment of subtalar osteoarthritis.

METHODSFrom March 2009 to September 2012, a total of 22 cases of old calcaneus fractures with subtalar osteoarthritis were treated with modified limited "L" incision and distraction bone block arthrodesis including 13 males and 9 females with a mean age of 35.3 years old (ranged 22 to 49). The mean time from calcaneal fracture was 21 months (ranged 11 to 32). According to the Stephens-Sanders classification, 16 cases were type II and 6 were type III. The modified-AOFAS ankle-hindfoot score was used for functional outcomes evaluation.

RESULTSThere was one incision necrosis and no infection, implant failure, bone-graft absorbed or talus necrosis was note at the follow-up time. A total of 21 cases were followed up for a mean time of 29 months (ranged from 18 to 46 months). All of the cases reached a bony union within 4 months postoperation. The mean modified-AOFAS ankle-hindfoot score was 82.6 points (ranged from 66 to 92 points),reached a significantly improvement in comparing with the mean preoperative score (50.8 points,ranged from 32 to 65 points, P < 0.01).

CONCLUSIONThe modified limited"L" incision beside the Achilles tendon with distraction bone block arthrodesis is an acceptable and alternative treatment method for subtalar osteoarthritis. This method is easy to use and with less complication. It can correct the main pathological changes and reach good functional outcomes.

Adult ; Arthrodesis ; methods ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis ; surgery ; Subtalar Joint ; surgery

Objective To investigate the influence of citalopram on the fast response action potential,slow response action potential,in vitro electrocardiogram(ECG) and in vivo ECG of cardiac myocytes,and explore its mechanism of adverse cardiac effects. Methods Conventional microelectrode technique was employed to record the fast and slow response action potentials of the isolated papillary muscles of guinea pigs.In vivo and in vitro ECG were recorded from anesthetized animals and Langendorff-perfused hearts,respectively. Results Citalopram could prolong the RR interval and QRS duration of in vivo ECG.The premature ventricular contraction and atrial ventricular block were induced by 12.5?10-6 mol/L citalopram.The maximum ascending velocity of 0 phase(Vmax),action potential amplitude(APA) and action potential duration(APD50 and APD90) were dose-dependently decreased by citalopram in the fast and slow response action potentials of guinea pigs,respectively. Conclusion Citalopram can inhibit sodium and calcium channels effectively,which may be the ionic mechanism that citalopram induces arrhythmia in the clinical practice.

OBJECTIVETo study the physiologic indexes, yield and the contents of alkaloids of Fritillaria cirrhosa D. Don under different temperature.

METHODThe growth temperatures (15, 20, 25, 30 degrees C) of F. cirrhosa were controlled by using artificial climate, the growth was observed, the contents of chlorophyll a and b, soluble sugar, MAD, proline of the leaves of F. cirrhosa were tested, and the yield and the alkaloids content of the bulbs were analyzed.

RESULTThe growth period of F. cirrhosa under 15, 20 degrees C were appropriately extended. The difference of the content of leaves chlorophyll b under four temperatures and the contents of total chlorophyll and chlorophyll a under 15, 20, 30 degrees C were not significant. The contents of soluble sugar, MAD and proline of leaves and the growth ratio, dry weight and content of alkaloids of bulb increased with the temperature decrease.

CONCLUSIONHigher temperature is not suitable for the growth of F. cirrhosa. Under the relatively lower temperature, the growth period of F. cirrhosa extended, the bulb can grow properly, and the content of alkaloid increased. F. cirrhosa can improve its cold tolerance by increasing the content of proline and soluble sugar, and it also can maintain the normal content of chlorophyll under the lower temperature.

Alkaloids ; metabolism ; Fritillaria ; growth & development ; metabolism ; Gene Expression Regulation, Plant ; Temperature

This study was aimed to investigate the protective effect of Wit3a gene modification on mouse bone marrow mesenchymal stem cells against the injury induced by Ara-C. The gene-modified MSC steadily expressing Wnt3a were established by adenovirus system. The acute direct damage effects of different concentrations of Ara-C on the unmodified MSC and the gene-modified MSC were assessed by using an in vitro culture system, and the corresponding controls were set. The proliferation and apoptosis of MSC exposed to Ara-C were detected by cell count kit-8 (CCK-8) and flow cytometry. The expression of BCL-2 protein related with cell apoptosis was assayed by Western blot. The results indicated that as compared with unmodified MSC, Ara-C exhibited a less inhibitory effect on the proliferation of gene-modified MSC. There was obvious difference between unmodified MSC and gene-modified MSC (p < 0.05). The proliferation of gene-modified MSC began to recover at 72 hours after removal of Ara-C. However, unmodified MSC showed sustained suppression of proliferation after withdrawal of Ara-C. In apoptosis, the apoptosis rate of gene-modified MSC induced by Ara-C was significantly lower than those of unmodified MSC (p < 0.05). In addition, the expression levels of BCL-2 protein in gene-modified MSC were up-regulated compared with unmodified MSC (p < 0.05). It is concluded that Wnt3a gene modification can significantly mitigate the damage of mouse bone marrow MSC induced by Ara-C.
Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Cytarabine ; adverse effects ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Mice ; Organisms, Genetically Modified ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Wnt3A Protein ; genetics

OBJECTIVETo investigate the effect of diet-induced obesity on the apoptosis of testicular spermatogenic cells in pubertal male rats.

METHODSForty healthy male rats were equally and randomly divided into a control and a high-fat group, the former fed on normal diet, while the latter high-fat and high-calorie diet. The testes of the rats were harvested at the end of 10 weeks for detection of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) in the peripheral blood with the automatic biochemical analyzer. Pathological changes of the testis were observed under the light microscope, the apoptosis of the testicular cells detected by TUNEL, the expressions of Bcl-2 and Bax proteins determined by immunohistochemistry, and those of Bcl-2 mRNA and Bax mRNA measured by RT-PCR.

RESULTSThe levels of TC, TG, LDL-C and HDL-C were significantly higher in the high-fat group (5.17 +/- 0.17, 1.18 +/- 0.09, 1.76 +/- 0.11 and 5.08 +/- 0.18) than in the control (1.38 +/- 0.12, 0.39 +/- 0.05, 0.97 +/- 0.07 and 0.75 +/- 0.06) (P < 0.05), so was the apoptotic index of spermatogenic cells (37.17 +/- 2.74 versus 5.16 +/- 0.81, P < 0.01), and the apoptotic spermatogenic cells were mainly spermatogonia and spermatocytes. The expressions of Bax protein and Bax mRNA were markedly higher in the high-fat group (153.26 +/- 8.74 and 1.08 +/- 0.12) than in the control (101.81 +/- 6.14 and 0.37 +/- 0.04) (P < 0.01), while those of Bcl-2 protein and Bcl-2 mRNA remarkably lower in the former (139.26 +/- 7.21 and 0.46 +/- 0.05) than in the latter (159.37 +/- 8.96 and 1.05 +/- 0.11) (P < 0.01).

CONCLUSIONDiet-induced obesity can increase the apoptosis of spermatogenic cells in the rat testis, which may be associated with the reduced expression of Bcl-2 and elevated expression of Bax.

Animals ; Apoptosis ; Diet ; Lipids ; blood ; Male ; Obesity ; etiology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Testis ; cytology ; pathology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo observe the effect of Wnt3a-transduced mouse bone marrow mesenchymal stem cells (MSC) on the proliferation of T lymphocytes.

METHODSMSC were isolated from C57BL/6 mouse bone marrow and expanded in vitro, then identified by flow cytometry and their differentiation capacity into osteocytes and adipocytes were determined. Recombinant plasmids containing Wnt3a gene, were transfected with lipofectamine into HEK293 cells by the AdEasy system. Viral particles were collected to infect MSC and adenovirus vector expressing GFP (Ad-GFP) was used as control. The expression of GFP in MSC was observed using fluorescence microscopy and the protein levels of Wnt3a and β-catenin were determined by Western blot. Wnt3a-transduced and Ad-GFP transduced MSC were separately cocultured with spleen lymphocytes stimulated by ConA, at the ratio of 1:100, 1:50 or 1:10 respectively. The proliferation rate of T lymphocytes was estimated by Cell Cout Kit-8 (CCK-8) and the level of cytokine by ELISA.

RESULTSFCM analysis showed that the MSC were highly positive for CD90.2, CD44 and negative for CD34, CD45, they could differentiate into osteoblasts and adipocytes after induction; The titer of recombinant adenoviruses was up to 1 × 10(10) pfu/ml. After infected with the adenoviruses, MSC had the strongest GFP expression at 72 h and the efficiency of infection was 50%-60%. The expressions of Wnt3a and β-catenin protein in the Wnt3a-transduced MSC were significantly increased. MSC could suppress the proliferation of T lymphocytes in a dose-dependent manner. When MSC cocultured with spleen lymphocytes at 1:10 ratio, T lymphocyte proliferation rate and the level of IFN-γ were (55.41 ± 1.75)% and (326.70 ± 14.41) pg/ml respectively in Ad-GFP transduced MSC group, while in Wnt3a-transduced MSC group, they were (37.27 ± 2.66)% and (218.80 ± 12.93) pg/ml respectively. There was no effect on the production of IL-2.

CONCLUSIONCompared to Ad-GFP transduced MSC, Wnt3a-transduced MSC exhibit a more potent inhibitory effect on the proliferation of T lymphocytes.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Proliferation ; Female ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; cytology ; Transduction, Genetic ; methods ; Wnt3A Protein ; genetics ; metabolism

OBJECTIVETo explore the value of a novel prenatal diagnosis model using combined chromosomal karyotyping and BACs-on-Beads(BoBs), a newly-developed technique.

METHODS1048 single pregnancy pregnant women with various diagnostic indications were performed amniocentesis for prenatal diagnosis with karyotyping and BoBs simultaneously.

RESULTSAmong 1047 successfully cultured specimens, 50 chromosomal abnormalities were identified with BoBs, including 43 common chromosomal trisomies, 3 chimeric chromosomes and 4 structural abnormalities, of which 3 microdeletions/microduplications were not detected with karyotyping. Except for extra yield of 1 Robertsonian translocation, the other numerical chromosomal abnormalities were detected with both karyotyping and BoBs. Ten fetal chromosome abnormalities were confirmed with karyotyping, including 8 structural abnormalities and 2 chimeric chromosomes.

CONCLUSIONCombination of karyotyping and BoBs turned out to be a rapid, complementary and effective diagnostic model for fetal chromosomal abnormalities and microdeletion syndromes, which could yield a higher detection rate of fetal chromosomal abnormalities and chromosomal microdeletions/microduplications.

Adult ; Chromosome Aberrations ; Chromosome Disorders ; diagnosis ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Testing ; methods ; Humans ; Karyotyping ; instrumentation ; methods ; Male ; Pregnancy ; Prenatal Diagnosis ; instrumentation ; methods ; Young Adult

Objective To explore the value of BACs-on-BeadsTM(BoBs)technique in detection of chromosome microdeletions/microduplications.Methods Samples for detection included 1 486 amniotic fluid samples for prenatal diagnosis and 7 peripheral blood samples with suspected chromosome microdeletions/microduplications collected in Zibo Maternal and Child Health Hospital from March 2015 to January 2017.BoBs and karyotyping were performed for all samples and SNP arrays were carried out to confirm the positive results of BoBs.Results 1 case of DiGeorge syndrome, 3cases of 22q11 microduplication syndrome,and 3 cases of Williams-Beuren syndrome were detected by BoBs,among which 1 case of Williams-Beuren syndromewas detected from peripheral blood and the rest were detected from amniotic fluid samples of high-risk gravidas after serological screening.None of the above 7 cases with chromosome microdeletion/microduplication syndromes was detected by karyotyping,butall of them were identified by SNP arrays.Conclusion BoBstechnique is capable of detecting specific chromosome microdeletions/microduplications with high accuracy and efficiency, which couldbe an auxiliary diagnostic tool for fetal chromosome abnormalities in addition to karyotyping.

ObjectiveTo investigate the difference in protein expression between hepatocellular carcinoma (HCC) patients with recurrence and those with good prognosis, the differential expression and regulatory mechanism of miR-152-3p target proteins, and the role of miR-152-3p in the recurrence of HCC. MethodsTMT-labeled proteomic sequencing and RT-PCR were used to measure the expression of proteins and the expression of miR-152-3p in the HCC tissue of six patients with recurrence at 2 years after HCC resection and six patients with good prognosis at 5 years. Six databases were used to analyze the target genes of miR-152-3p, and Gene Ontology, DAVID, and REACTOME databases were used to perform target gene screening, enrichment annotation, and signal transduction pathway enrichment analysis. Gene mutation frequency and survival curve analysis were performed for the target genes of miR-152-3p to verify the role of miR-152-3p target genes in patients with HCC recurrence. The independent samples t-test was used for comparison of continuous data between two groups, and a Kaplan-Meier analysis was performed to investigate the survival rates of liver-related genes. ResultsCompared with the patients with HCC recurrence, the patients with good prognosis after HCC resection had a significantly higher transcriptional expression level of miR-152-3p in HCC tissue (P＜0.05). The results of protein sequencing showed that there were 365 differentially expressed proteins in HCC tissue between the patients with good prognosis and the patients with recurrence, and the analysis of HCC recurrence databases showed that 17 proteins were regulated by miR-152-3p. Further analysis of the signaling pathways showed that the function of the 17 target genes regulated by miR-152-3p was enriched in the translation and regulation of mitochondria and ribosome, and multiple enrichment revealed that six target genes were closely associated with mitochondrial respiratory chain complex, i.e., AKAP1, FOXRED1, MRPL28, MRPL50, SHC1, and STAU1. Gene mutation frequency and survival curve analysis showed that the loss or weakening of the function of mitochondrial respiratory chain-related target proteins seriously affected the prognosis and survival rate of patients. ConclusionThere is a significant difference in the expression of miR-152-3p in HCC tissue between patients with good prognosis and those with recurrence after HCC resection, and miR-152-3p may lead to the recurrence of HCC by regulating the target genes AKAP1, FOXRED1, MRPL28, MRPL50, SHC1, and STAU1, acting on the mitochondrial respiratory chain, and affecting the oxidative respiratory function of cells.

Objective To compare the efficacy and complications of the extend endoscopic endonasal approach (EEA) and open transcranial approach for resection of craniopharyngiomas. Methods The clinical data from 46 patients with craniopharyngiomas with extend EEA and 54 patients with transcranial route in our department was analyzed retrospectively. The gross total resection (GTR) rate,length of hospital stays and complications of the two groups were compared. Results The tumor diameters of were larger in the endoscopic group than in the transcranial group (3.5 ± 1.3cm vs. 3.0±0.8 cm, P<0.05). The endoscopic group had a greater GTR rate (67.4％vs. 46.3％, P<0.05)and improved visual outcome(84.2％ vs. 59.5％,P<0.05),but lower rate of hypopituitarism (56.5％ vs. 75.9％,P<0.05)and permanent diabetes insipidus (51.4％ vs.72.7％,P<0.05). On the contrast, the endoscopic group had a greater rate of cerebrospinal fluid leak(4.3％ vs. 0.0％,P>0.05)and longer hospital stays(17.0±3.6 d vs. 13.1±2.3 d,P<0.01). Hyposmia(34.8％)and hemorrhinia (2.2％)only happened in the endoscopic group. Conclusion Compared with transcranial route, the extend EEA for craniopharyngiomas is minimal invasion and effective, which can effectively improve the GTR rate and reduce the clinical symptoms.