Objective:To observe clinical and pathological features of pancreatic solid-pseudopapillary tumor ( SPPT) , and to find some useful immunohistochemical methods for its differential diagnosis.Methods: The clinical features of 14 SPPT patients were obtained. Each case underwent microscopic observation and immunohistochemical staining. The primary antibodies were CgA, Syn, E-cadherin, β-catenin and Cyclin Dl. These results were compared with 5 pancreatic well-differentiated tumors and well-differentiated carcinomas ( WET/WEC). Results: SPPT mainly involved young women, and the head of pancreas was the commonest location. Tumors were always in solid and cystic gross appearance.Although the tumor' s borderlines seemed clear, focal infiltrations could often be identified. The histological features of SPPT were similar in some aspects to those of WET/WEC, especially the solid pattern of WET/WEC. Both of them could express CgA and Syn. But all SPPTs lost E-cadherin membranous signals, and even had some nuclear signals (5/14) , while all WET/WECs remained the same staining pattern with normal pancreas cells, β-catenin positive signals in SPPTs were located both in nuclei and plas mas. WET/WECs' positive signals were all in membranes and plasmas, but negative ones in nuclei. Perinuclear dot-like signals could also be seen in the majority cells, which were similar to normal islet cells' staining pattern. SPPTs' nuclear positive rates of Cyclin Dl were usually more than 70% (12/14). WET/WECs' rates were all lower than 30%. Conclusion: Comprehensive analysis of patients'clinical, pathological features and immunohistoehemistry results, including E-cadherin, β-catenin and Cyclin Dl, was helpful to the diagnosis of SPPT and its differential diagnosis of WET/WEC.

Objective To study the EEG discharge index, intelligence test and event-related potential P300 in BECT, and to analyze the change of EEG discharge index and cognitive function before and after the treatment.Methods Sixty patients with BECT were enrolled in this study, they were treated with EEG, intelligence tests and P300 before and after the treatment.Results (1) The EEG discharge index were reduced remarkly after treatment in BECT with levetiracetam and lamotrigine, the difference was statistically significant (P ＜ 0.05).(2) Comparing before and after 3 or 6 months treatment, the latency of P300 had reduced with significant difference (P ＜ 0.05).(3) After 3 months treatment, VIQ and FIQ has no obvious improvement, but PIQ has improved.After 6 months treatment, VIQ、 PIQ and FIQ were improved.The difference was statistically significant (P ＜ 0.05).(4) There was a negative correlation of EEG P300 latency (r =0.175), as well as there was a negative correlation between EEG discharge index and intelligence test (r =0.044).Conclusion There is impaired cognitive function in BECT, especially the more frequently the EEG discharge, the more extended of P300 latency, as well as the more serious damage of intelligence and cognition after treatment.The intelligence were improved after treatment with Levetiracetam and lamotrigine, the longer the treatment time, the more obvious of intelligence levels improve.

OBJECTIVETo investigate the mechanism that the formulas for activating blood and resolving stasis can regulate hemopoietic stem cell to produce new blood.

METHODRats were established animal model of acute cerebral infarction by referencing Olivette' method. They were randomly divided into model group, the group of the high, middle, low dose of the formulas for activating blood and resolving stasis. Each group and then wasrandomly divided into subgroups by 1, 3, 7, 14, 28 d. Xuesaitong capsule was formulated into 20, 40, 60 g x L(-1) with normal saline. The rats were given gavage drugs once a day until the experient ended, and the model group was administrated by intragastrical perfusion of normal saline. ELISA was used to detect the expression of SCF in peripheral blood and bone marrow among different groups at different time points. Flow cytometry was used to observe the changes of CD117 in blood and bone marrow.

RESULTThe CD117+ HSC and SCF concentration in peripheral blood and bone marrow of model group were increasing during 1-14 d,there was a peak on the 14th day, then the expression was reducing. CD117+ HSC and SCF concentration rising trend in the group of the high, middle dose of the formulas for activating blood and resolving stasis was preceded model group (P < 0.05).

CONCLUSIONActivating blood and resolving stasis can regulate hemopoietic stem cell to produce new blood, and it is through the regulation of CD117+ HSC number to achieve the purpose.

Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Capsules ; Cerebral Infarction ; blood ; drug therapy ; genetics ; metabolism ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; Hematopoietic Stem Cells ; drug effects ; metabolism ; Humans ; Male ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cell Factor ; genetics ; metabolism

OBJECTIVETo compare therapeutic effects between unilateral decompression technique only and open decompression technique with fusion and internal fixation for the treatment of lumbar spinal stenosis.

METHODSFrom March 2008 to February 2011, 82 patients with lumbar spinal stenosis were treated with operations, and divided into two groups. There were 13 males and 19 females in group A, with a mean age of (56.31±4.31) years old. The patients in group A were treated with unilateral decompression via fenestration technique only, including 23 patients obtaining single level decompression and 9 patients obtaining two levels decompression. In group B, there were 18 males and 32 females, with a mean age of (57.53±4.28) years old. The patients in group B were treated with open decompressive technique with fusion and internal fixation, including 38 patients obtaining single level decompression and 12 patients obtaining two levels decompression. The VAS of back pain and leg pain, ODI were recorded before and after surgery to evaluate low back pain,leg pain and walking tolerance.

RESULTSAll the patients were followed up, and the duration ranged from 10.9 to 43.4 months,with a mean of 32.8 months. There were no differences in age, stenosis level, VAS of back and leg pain and ODI before surgery between two groups. Compared with the corresponding ones in group B, the operation time, blood loss, hospitalization time,recovery time of routine daily life and finacial expenditure of patients were all shorter or less in group A. There was no statistically difference in complications between two groups.

CONCLUSION"Unilateral decompression via fenestration technique" is a less invasive and more effective decompressive technique for degenerative spinal stenosis without posterior elements damage. It has advantages in operation time, blood loss, hospitalization time, recovery to daily life and financial expenditure. When controlling the operative indications strictly, the technique could be an important procedure for surgical treatment of degenerative spinal stenosis, especially in the elderly population.

Adult ; Aged ; Case-Control Studies ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery

AC.

AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.

Objective To investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1α level of HDPC supernatants stimulated by lipopolysaccharide(LPS)and tumor necrosis factor-α(TNF-α),and to explore the role of SDF-1αon the proliferation and the migration of HDPC.Methods The expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique.The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-α of difierent concentrations for 48h and then the SDF-1α level was assayed by quantitative sandwich ELISA.Meanwhile,the effects of recombinant human SDF-1α(rhSDF-1α)on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.Results CXCR4 was expressed in cytomembrane of HDPC and SDF-1α was secreted into their normal cell supematants with a concentration of(4513.55±962.92)ng/L. The secretion of SDF-1α was both significantly decreased by stimulation with LPS and TNF-α(P＜0.05).In addition,rhSDF-1α stimulated the HDPC proliferation at the concentrations of 50,100,200μg/L(P＜0.01)and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50,100μg/L(P＜0.05).Conclusions SDF-1α accelerated the proliferation and the migration of HDPC which expressed CXCR4.SDF-1-CXCR4 axis may play a role in repair of pulp injury.

Objective:By observing the changes of hyperthermia on serum carbohydrate antigen 125(CA125),human epididymal protein 4(HE4),chitinase protein(YKL) and immune function index effects,to investigate its effect on cancerous ascites and its antitumor effect of cytoreductive surgery combined with intraperitoneal hyperthermic perfusion chemotherapy and in elderly patients with advanced ovarian cancer.Methods: A total of 72 elderly patients with advanced ovarian cancer were enrolled in the study.They were randomly divided into contrast treatment group and combined treatment group,36 cases in each group.The patients in contrast treatment group were treated with cytoreductive surgery combined with intraperitoneal hyperthermic perfusion chemotherapy.The combined treatment group was treated with partial radiofrequency hyperthermia on the basis of the treatment of the control group.The efficacy was evaluated after 2 courses.The changes of CA125,HE4 and YKL contents and T cell subsets were compared between the two groups before and after treatment,and the safety was evaluated.Results: There was no difference in the operation between the two groups.The total effective rate was 80.0% in combined treatment group,which was significantly better than 55.6% in contrast treatment group(χ2=5.175,P<0.01).The effective rate of ascites control was 80.6% in combined treatment group,which was significantly higher than that in contrast treatment group 52.4%(χ2=3.962,P<0.05).After treatment,the levels of CA125,HE4 and YKL in the two groups were significantly lower than those before treatment,and the decrease of the combined treatment group was more significant than contrast treatment group(P<0.05).After treatment,the levels of CD3+,CD4+,CD4+/CD8+in both groups were lower than those before treatment,CD8+was lower than that before treatment,and the improvement of the indexes in combined treatment group was more significant than contrast treatment group(P<0.05).There was no significant difference between the two groups in the adverse effects of bone marrow suppression,gastrointestinal reaction and liver and kidney dysfunction(P> 0.05).Conclusion: CRS combined with IPHC and hyperthermia can significantly improve the immunosuppression of ovarian cancer,reduce the level of CA125,HE4,YKL, improve clinical efficacy of elderly patients with advanced ovarian cancer.

BACKGROUNDMutations in fumarylacetoacetate hydrolase (FAH) gene can lead to tyrosinemia type 1 (HT1), a relatively rare autosomal recessive disorder. To date, no molecular genetic defects of HT1 in China have been described. We investigated a Chinese family with a HT1 child to identify mutations in FAH.

METHODSDNA sequencing was used for mutations screening in FAH gene. Real-time polymerase chain reaction (PCR) was performed to determine the FAH gene expression level. To confirm the presence of degradation by the nonsense-mediated mRNA decay pathway (NMD), the fragments containing R237X mutations were analyzed by primer introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and cDNA sequencing. Finally, the effects of the mutations reported in this study were predicted by online softwares.

RESULTSA boy aged 3 years and 8 months was diagnosed clinically with HT1 based on his manifestations and biochemical abnormalities. Screening of FAH gene revealed two heterozygous mutations R237X and L375P transmitted from his mother and father respectively. In this pedigree, the amount of FAH mRNA relative to a healthy control was 0.44 for the patient, 0.77 for his mother and 1.07 for his father. Moreover, both PIRA-PCR and cDNA sequencing showed significant reduction of the FAH mRNA with R237X nonsense mutation. The missense mutation of L375P was not reported previously and prediction software showed that this mutation decreased the stability of protein structure and affected protein function.

CONCLUSIONSThis is the first case of HT1 analyzed by molecular genetics in China. The R237X mutation in FAH down- regulates the FAH gene expression, and the L375P mutation perhaps interrupts the secondary structure of FAH protein.

Child, Preschool ; China ; Humans ; Hydrolases ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; genetics ; Nonsense Mediated mRNA Decay ; genetics ; Real-Time Polymerase Chain Reaction ; Tyrosinemias ; genetics

AIMTo detect the expression of HSP25 in rat dental follicles both in vivo and vitro, and explore the underlying mechanism of HSP25 on the proliferation and differentiation of rat dental follicle cells (DFCs).

METHODOLOGYImmunohistochemistry was performed to detect the expression of HSP25 in mandibles of postnatal rats on days 1, 3, 5, 7, 9 and 11 in vivo. In vitro, the expression of HSP25 in DFCs was detected by an indirect immunofluorescence assay. Thiazolyl blue tetrazolium bromide (MTT) assay, flow cytometry and alkaline phosphatase (ALP) assay were used to identify the time-course effect mediated by different concentrations of recombinant murine HSP25 of 0, 1, 10, 50 and 100 ng/mL on rat DFCs.

RESULTSExpression of HSP25 was not detected in dental follicles of the rats until day 5 after birth, but became up-regulated in a time-dependent manner till day 11. HSP25 was detected in the cytoplasm of cultured rat DFCs. No significant difference could be observed in the proliferation of DFCs after stimulation with different concentrations of HSP25 on days 1, 2 and 3 (P > 0.05). HSP25 at concentrations of 50 ng/mL and 100 ng/mL up-regulated the ALP activity of DFCs on day 9 (P < 0.05).

CONCLUSIONHSP25-immunoreactivity increased chronologically during the development of dental follicles. The protein had no significant effect on cell proliferation but may play a role in cementoblast/osteoblast differentiation of DFCs.

Alkaline Phosphatase ; analysis ; Ameloblasts ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Coloring Agents ; Cytoplasm ; ultrastructure ; Dental Sac ; cytology ; growth & development ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; HSP27 Heat-Shock Proteins ; analysis ; physiology ; Odontoblasts ; cytology ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; Thiazoles ; Tooth Germ ; cytology ; Up-Regulation ; physiology