AIM: To establish a fluorogenic quantitative polymerase chain reaction (FQ-PCR) method for the routine examination of c-erbB-2 gene expression in breast cancer. METHODS: The c-erbB-2 standard gene was obtained by in vitro amplification of cloned c-erbB-2 fragment in plasmid PGEM-T easy vector. FQ-PCR product was detected by using a 7700 ABI PRISM sequence detector system and c-erbB-2 standard curve was obtained to quantity c-erbB-2 in unknown samples. RESULTS: “S” kinetics curve of FQ-PCR amplification was generated by relating the fluorescence signal intensity (△Rn) to the cycle number. The standard curve of c-erbB-2 was constructed by the linear relationship between the cycle threshold (ct) and the log of starting copy number. The high correlation (0.999) revealed the reliability of FQ-PCR. CONCLUSION: The FQ-PCR is a rapid, sensitive, reliable method for quantity of c-erbB-2 gene expression.

Experiments were conducted to investigate the suitability of the multistage in-situ reaction analyzer based on a micro fluidized bed ( MFB-MIRA) for measuring the rapid change of the gas concentration during gas-solid reactions. The results showed that the control performance of capillary temperature had a great impact on the stability of on-line measurement. Based on the observed regular patterns, the capillary temperature control system was equipped with a precision temperature controller. The control precision of capillary temperature reached ± 0 . 2℃, which guaranteed the high stabilities of the sampling flow rate and the chamber vacuum. The measured results of the modified gas monitoring system showed the periodic fluctuations of the on-line measurement were eliminated. The stability of measurement was significantly improved. The fluctuating range and relative standard deviation of the measured response to O2 in air changed from 1. 9% and 0. 5% to 1. 4% and 0. 2%, respectively. A pressure regulating device was also developed to control the absolute pressure at the gas sampling point. The control precision reached ± 0. 02 kPa. The measured results showed that the response of the process mass spectrometer was positively correlated with the absolute pressure at the sampling point, indicating the necessity of the pressure regulating device. The accuracy and repeatability of process mass spectrometer were improved. This study has enhanced the suitability of MFB-MIRA for studying rapid gas-solid reactions and broadened the scope of reliable applications of MFB-MIRA and process mass spectrometer.

Objective To retrospectivel y analyze the abdominal CT findings and pathological results of the chronic schist osomiasis so as to improve the diagnostic accuracy of the disease. M ethods The plain abdominal CT scanning was performed in 103 cases an d enhanced CT scanning in 81 cases. The pathological specimen which was consist ent with the section of CT scan was obtained in each cases. Results On CT scanning, liver cirrhosis was seen in 84 cases, various calci fication in liver in 71 cases, liver cancer in 12 cases, enlargement of sple en in 78 cases, calcification in spleen in 13 cases, wall-thickening in colon i n 27 cases, calcification in colon in 31 cases, and colon cancer in 9 cases. Pa thological examination revealed various fibrosis and formation of pseudolobule. The eggs and calcification could be seen in pseudolobule and septa, colonic sub mucosa, and regional lymph nodes. Fibrous hyperplasia in colonic wall and hyper plasia in mucous membrane were obvious. Fibrous hyperplasia and calcification w ere seen in spleen, but the eggs were not found. Conclusion The liver and colon are the major organs affected by chronic schistosomias is in abdomen, and the CT findings are obvious too. The pathological features o f spleen are accompanied with liver cirrhosis. CT is the important imaging meth od in diagnosing chronic schistosomiasis and pathological changes.

OBJECTIVETo evaluate the effectiveness of locking plate external fixator in treating middle and distal tibial fractures.

METHODSFrom January 2010 to January 2013,18 patients suffered from middle and distal tibial fractures were treated by locking plate external fixator,including 11 males and 7 females, with an average age of 53.5 (ranged from 13 to 80) years old,the course of disease ranged from 2 h to 3 d. According to AO classification, 4 cases were type A,11 cases were type B and 3 were type C. Among them,6 patients were open fracture, including 2 cases with type I, 3 cases with type II and 1 case with type III, according to Gustilo classification), 12 patients were close fracture. Operation time, postoperative complications were observed, and Johner-Wruhs scoring were used to evaluate clinical outcomes.

RESULTSAll patients were followed up from 6 to 15 (meaned 11) months. Two cases occurred skin necrosis (1 case occurred bone exposure), 2 cases occurred delayed union (all were open fracture), and 1 case occurred nail infection. No screw loosening or broken occurred. According to Johner-Wruhs scoring, 10 cases obtained excellent result,6 cases good,and 2 cases fine.

CONCLUSIONLocking plate external fixator for the treatment of middle and distal tibial fractures, which has advantages of lessen damage, shorter operative time, less complications and rapid functional recovery, is one of good choice.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Plates ; utilization ; External Fixators ; utilization ; Female ; Fracture Fixation ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P＜0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P＞0.05),but these parameters significantly changed before and after HVHF (P＜0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P＜0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVETo construct a convenient, reliable and visual model of hair follicle development to test the hair-inductive potential of follicular cells and investigate the molecular mechanism regulating hair follicle morphogenesis and cycling.

METHODSAn open chamber was transplanted into the nude mice dorsal skin, dermal and epidermal cells isolated from newborn C57BL/6 mice skin were mixed at a specific ratio and then injected into the chamber together, 1 week after transplantation, the chamber was removed, and then, hair formation and regeneration after hair plucking was observed.

RESULTS1 week after cells implantation, the wound was moist without apparent contraction and among that pink and translucent tissue was formed. 2 weeks after implantation, the wound healed completely. 3 weeks after implantation, black hair grew from the skin was observed. 4 weeks after implantation, thick and black hair grew from the skin vertically. Completely developed structure of hair follicle was observed with paraffin section and HE staining. 1 week after plucking, new hair had regrown. The ratio of cell component was varied, whereas the other component was fixed at 1 x 10(7) cells. When the number of epidermal cells was reduced to 1 x 10(6) cells, the efficiency of hair follicle reconstitution was mostly unchanged. On the other hand, the density of newly formed hair was diminished considerably by reducing the number of dermal cells to 5 x 10(6) cells or lower. Neither epidermal cells nor dermal cells transplanted alone formed hair follicle.

CONCLUSIONSNewborn mice skin cells transplanted by chamber method can construct a complete model of hair follicle development, which can be used to test the hair-inductive potential of follicular cells and investigate the molecular mechanism regulating hair follicle morphogenesis and cycling.

Animals ; Cells, Cultured ; Hair ; physiology ; Hair Follicle ; physiology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Nude ; Regeneration ; Skin ; cytology

The growth inhibitory effects of D-glucosamine hydrochloride (GlcNH(2).HCl), D-glucosamine (GlcNH(2)) and N-acetyl glucosamine (NAG) on human hepatoma SMMC-7721 cells in vitro were investigated. The results showed that GlcNH(2).HCl and GlcNH(2) resulted in a concentration-dependent reduction in hepatoma cell growth as measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. This effect was accompanied by a marked increase in the proportion of S cells as analyzed by flow cytometry. In addition, human hepatoma SMMC-7721 cells treated with GlcNH(2).HCl resulted in the induction of apoptosis as assayed qualitatively by agarose gel electrophoresis. NAG could not inhibit the proliferation of SMMC-7721 cells. GlcNH(2).HCl exhibited antitumor activity against Sarcoma 180 in Kunming mice at dosage of 125-500 mg/kg, dose of 250 mg/kg being the best. GlcNH(2).HCl at dose of 250 mg/kg could enhance significantly the thymus index, and spleen index and could promote T lymphocyte proliferation induced by ConA. The antitumor effect of GlcNH(2).HCl is probably host-mediated and cytocidal.
Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA ; metabolism ; DNA Fragmentation ; drug effects ; Glucosamine ; analogs & derivatives ; therapeutic use ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Male ; Mice ; Sarcoma 180 ; drug therapy

OBJECTIVETo study the gene expression of transforming growth factor beta receptor II (TbetaR II) in pathological scar.

METHODSTwenty samples of pathological scar were collected from 20 burn or trauma patients hospitalized in the General Hospital of Ji'nan Military Command from 2007 to 2009. Twenty specimens of epidermal layer were obtained from the middle portion and the edge of pathological scars. Twenty normal skin specimens which were located more than 10 cm away from the lesion sites of 20 patients were collected as self-controls. Serum from 1-2 mL whole blood were obtained from each of the 20 patients for second self-control. Eight normal skin specimens from 8 patients without pathological scar, discarded from un-related operations, were also collected as negative-control. Positive expressions of TbetaR II in three different skin specimens were determined with biotin-streptavidin-peroxidase staining. Gene expressions of TbetaR II in all specimens were compared with PCR-single strand conformation polymorphism analysis and gene sequencing. Data were processed with Fisher's exact test.

RESULTSPositive expression of TbetaR II in pathological scar epidermis was lower than that in normal skin specimen of patients with pathological scar or normal skin specimen of patients without pathological scar, and TbetaR II was mainly located in the basal layer of epidermis. Positive expressions of TbetaR II were seldom found in acanthocytes, granular cells, and cuticle or even non-existing. No abnormality of TbetaR II was found in normal skin epidermis or serum samples of pathological scar patients or normal skin epidermis of patients without pathological scar. TbetaR II expressing in 8 specimens of epidermis of pathological scar showed abnormal electrophoresis pattern at poly A fragments hand and loss of one A base in DNA fragment (P = 0.044).

CONCLUSIONSThere may he abnormal gene expression of TbetaR II in pathological scar epidermis. Replantation of epidermis of scar may increase the risk of scar recurrence, while replantation of normal skin of patients with scar on wound may not increase the risk of scar recurrence.

Adolescent ; Adult ; Child ; Child, Preschool ; Cicatrix ; metabolism ; pathology ; Epidermis ; metabolism ; Female ; Gene Expression ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Receptors, Transforming Growth Factor beta ; genetics ; metabolism ; Young Adult

OBJECTIVETo analyze the surgical treatment result and clinical characteristics of hilar cholangiocarcinoma in order to improve the rate of early diagnosis and radical resection.

METHODSBetween 1986 and 2004,84 hilar cholangiocarcinoma patients underwent surgery, and their data were retrospectively reviewed.

RESULTSAccording to the Bismuth-Corlette staging system, 7 were type I, 18 type II, 22 type II a, 12 type IlI b, 20 type IV and 5 unclassified. 32 patients (38.1%) had had the history of operation for cholelithiasis before or were found to have cholelithiasis simultaneously at the time of diagnosis. The rate of making correct diagnosis by ultrasound, CT and MRCP was 71.4% , 84.0% and 91.4% , respectively. Of these 84 patients, 24 (28.6%) underwent radical resection, 14 (16.7%) palliative resection and 30 (35.7%) only internal or external drainage, while 16 patients was found to have contraindication for any further surgical intervention. The overall operation rate was 81.0% (68/84) with a radical resection rate of 35.3% (24/68). The 1-, 3- and 5-year survival rates was 70.8%, 50.0% and 20.8% in the radical resection group, and 50.0%, 21.4% and 0 in the palliative resection group, respectively. There was a statistically significant difference in the survival between two groups. Whereas in the internal or external drainage group, the 1-, 3- and 5-year survival rates was 20.0% ,10.0% and 0. All of the patients who did not undergo surgical intervention died within one year.

CONCLUSIONCholelithiasis may play an important role in the pathogenesis of hilar cholangiocarcinoma. Early diagnosis and radical resection are two important factors to improve the prognosis of hilar cholangiocarcinoma. Skeletonization of hepatoduodenal ligament with partial liver resection can improve the rate of radical resection for hilar cholangiocarcinoma.

Adenocarcinoma ; diagnosis ; surgery ; Adenocarcinoma, Mucinous ; diagnosis ; surgery ; Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; diagnosis ; surgery ; Bile Ducts, Intrahepatic ; Biliary Tract Surgical Procedures ; methods ; Cholangiocarcinoma ; diagnosis ; surgery ; Drainage ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Analysis