Carcinoma, Renal Cell ; immunology ; pathology ; surgery ; Humans ; Keratin-19 ; metabolism ; Kidney Neoplasms ; immunology ; pathology ; surgery ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Nephrectomy ; Vena Cava, Inferior ; pathology ; surgery

To investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 pathway and the growth of gastric cancer cell lines at different GHR expression status, the eukaryotic expression vector targeting human GHR (pGPU6/GFP/Neo-shGHR and pGPU6/GFP/Neo-scramble) was constructed and transfected into MGC803 cells by Lipofectamine 2000. Stable expressive cell lines were obtained by G418 screening. The expression of GHR was analyzed by Western blotting. After being stimulated with rhGH, cell growth was detected by MTT assay. Cell cycle and apoptosis were examined by flow cytometry. The components of JAK2/STAT3 signaling pathway were detected by Western blotting. There is no significant difference of GHR expression between MGC803 and pGPU6/GFP/Neo-scramble-transfected cells (named as MGC803-NC) (P > 0.05). Compared with MGC803, the GHR expression in pGPU6/GFP/Neo-shGHR-transfected cells (named as MGC803-shGHR) decreased significantly (protein decreased 50%). The cells were treated with rhGH at 0, 150 and 300 ng x mL(-1), the growth rate of MGC803 and MGC803-NC increased significantly, PI and the number of G2/M phase cells all increased significantly, and apoptosis decreased significantly. Western blotting revealed that the expression of pJAK2 and pSTAT3 was up-regulated after being treated with rhGH in MGC803 and MGC803-NC cells. In contrast, similar change was not observed in MGC803-shGHR cells. Knockdown of GHR gene may decrease the sensitivity of gastric cancer cells to rhGH, and down-regulating of components of the expression of JAK2/STAT3 signaling pathway may be the potential mechanisms.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Human Growth Hormone ; genetics ; pharmacology ; Humans ; Janus Kinase 2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Somatotropin ; genetics ; metabolism ; Recombinant Proteins ; genetics ; pharmacology ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury. This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups (1, 3, and 10 mg·kg^-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues. DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.

OBJECTIVETo investigate the diagnostic pathological features of endometrial carcinomas present in women under 40 years of age (by curettage samples).

METHODSA retrospective analysis was performed on 20 cases of endometrial carcinomas in women under 40 years of age.

RESULTSThe patients included 18 endometrioid adenocarcinoma cases, one adenosquamous carcinoma case, and one papillary serous carcinoma case. The morphological features of the endometrial adenocarcinoaare loss of polarity (orientation) of the endometrial glands. The tumor cells have large round vesicular nuclei, prominent nucleoli and coarse chromatin. In addition, endometrial stroma was instead of by the fibrous and/or granulation tissue type stroma, usually with inflammatory response. Majority cases of endometrioid adenocarcinomas had superficial invasion of myometrium and no lymph node metastasis. The adenosquamous case metastasized to the ovaries and the papillary serous carcinoma case metastasized to the pelvic lymph nodes.

CONCLUSIONSEndometrial carcinomas which are highly differentiated endometrioid adenocarcinomas occur frequently in women under 40 years of age. An important differential diagnosis of the endometrioid carcinoma in a curettage specimen is to differentiate it from atypical endometrial hyperplasia and atypical polypoid adenomyoma.

Adenomyoma ; pathology ; Adult ; Carcinoma, Adenosquamous ; secondary ; Carcinoma, Endometrioid ; pathology ; Carcinoma, Papillary ; pathology ; Diagnosis, Differential ; Dilatation and Curettage ; Endometrial Hyperplasia ; pathology ; Endometrial Neoplasms ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Myometrium ; pathology ; Ovarian Neoplasms ; secondary ; Pelvis ; Retrospective Studies

OBJECTIVETo establish a solvent desorption Gas chromatographic method for detecting the isoflurane in air of workplaces.

METHODSThis method is based on "Standardization of methods for determination of toxic substances in workplace air".

RESULTSThis method presents the linear relation with the minimum detectable limit 1.0 µg/ml and the minimum detectable concentration 0.07 mg/m(3). The precision (RSD) was 0.5% ∼ 5.0%, the mean dsorption efficiencies were 96.7% ∼ 98.9%, the absorption efficiencies were 92.1% ∼ 100%, the breakthrough volume was 3.7 mg isoflurane/100 mg active carbon. Other volatile organic solvents (Sevoflurane, Enflurane and Ethyl Alcohol) did not interfere the detection. The sample could be stored in the active carbon tube at least for 10 days.

CONCLUSIONThis method is meet the requirement of GBZ/T 210.4-2008 "Guide for establishing occupational health standards-Part4: Determination methods of air chemicals in workplace" and is feasible for determining the isoflurane in the air of workplaces.

Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Isoflurane ; analysis ; Workplace

OBJECTIVETo investigate the effect and safety of the implantation of a new type of testicular prosthesis in the treatment of testis loss.

METHODSWe recruited for this study 18 patients with testis loss treated by testicular prosthesis implantation, including 10 cases of prostate cancer, 3 cases of anorchia, 2 case of orchiatrophy, 2 cases of hermaphroditism and 1 case of cryptorchidism. The prosthesis was a hollow silicone elastomer YH-G1 made in China, selected according to the volume of the scrotum and the size of the contralateral testis.

RESULTSThirteen of the patients received testicular prosthesis implantation with orchiectomy, and the other 5 underwent the procedure 6 months later. The operation time of testicular prosthesis implantation was (22.6 +/- 4.6) min, ranging from 15 to 30 minutes. All the patients were discharged after 12 hours of postoperative observation, with a mean hospital stay of (1.3 +/- 0.4) days. A follow-up after 6 months revealed no complications in 17 cases. Rejection occurred in 1 case at 3 months after the implantation, ending in the removal of the prosthesis. Of the 17 successful cases, 15 were very satisfied with the size of the prosthesis, 14 with its weight, 12 with its comfortableness, and all with the appearance of the scrotum and the position of the prosthesis, while 5 found the implant too rigid.

CONCLUSIONThe implantation of the new home-made silicone elastomer testicular prosthesis YH-G1 was safe and effective for the treatment of testis loss, and could meet the esthetic and psychological requirements of the patient. But further observation is needed for its long-term complications and influence on the patient's quality of life.

Adolescent ; Adult ; Aged ; Gonadal Dysgenesis, 46,XY ; surgery ; Humans ; Male ; Middle Aged ; Orchiectomy ; Patient Satisfaction ; Prostheses and Implants ; Prosthesis Implantation ; adverse effects ; Silicone Elastomers ; Testis ; abnormalities ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of survivin antisense oligodeoxynucleotide (ASODN) on proliferation and apoptosis of human malignant melanoma cells.

METHODShMMC A375 colonies in log growth phase were collected and divided into control group (C, without transfection), sense chain group [SC, transfected with 600 nmol/L survivin sense oligodeoxynucleotide (ODN)], mismatch chain group (MC, transfected with 600 nmol/L survivin mismatch sense ODN), liposome group (L, treated with liposome), antisense chain group (AC, transfected with survivin ASODN, and subdivided into AC 200, 400, 600 nmol/L subgroups) according to the random number table. Transfection result was observed under inverted fluorescence microscope. Inhibition rate of cell proliferation was calculated after determination of cell viability with MTT method. Cell cycle and apoptosis rate were detected with bi-variable flow cytometry. Expression of survivin protein was determined with Western blot. Activity of caspase-3 was assessed with kinase method. Data were processed with analysis of variance.

RESULTS(1) Cell transfection rates in SC, MC, AC 600 nmol/L groups were all above 80%. (2) Compared with those in SC group [(5.23 +/- 0.25)%], MC group [(5.09 +/- 0.13)%] and L group [(4.70 +/- 0.45)%], inhibition rates of cell proliferation in AC 200, 400, 600 nmol/L groups 24 hours after transfection [(10.30 +/- 0.56)%, (16.69 +/- 0.58)%, (24.67 +/- 0.67)%] were significantly increased (F = 746.91, and P values all below 0.05). As time after transfection went on, proliferation inhibition rate was increased obviously. (3) Apoptosis rate in AC 200, 400, 600 nmol/L groups 24 hours after transfection was respectively (13.5 +/- 1.9)%, (20.1 +/- 1.5)%, (32.1 +/- 2.9)%, which were significantly higher than those in C, SC, MC, and L groups [(6.5 +/- 0.6)%, (5.6 +/- 0.7)%, (6.4 +/- 1.0)%, (6.5 +/- 1.3)%, F = 139.9, P values all below 0.05]. Cells in AC group were blocked in G2/M stage. (4) Compared with those in C group, expression amount of survivin protein decreased, and caspase-3 activity obviously increased (F = 63.1, P values all below 0.05) in AC group. No significant difference in caspase-3 activity between SC, MC, L groups and C group was observed (F = 0.512, P values all above 0.05).

CONCLUSIONSSurvivin ASODN can inhibit the proliferation of hMMC A375 in a concentration-time dependent manner, and it induces G2/M stage block and promotes its apoptosis.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Melanoma ; metabolism ; pathology ; Microtubule-Associated Proteins ; genetics ; pharmacology ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Transfection

BACKGROUNDChronic liver disease causes aberrant formation of fibrous tissue that impedes normal liver function, ultimately resulting in liver cirrhosis. Iron uptake can occur within the hepatic parenchyma or within the various nodules that form in a cirrhotic liver, termed siderotic nodules (SN). We aimed to investigate the diagnostic performance of susceptibility weighted imaging (SWI) for detection of SN in patients with liver cirrhosis, and to evaluate the potential of SN numbers for assessing the degree of hepatic iron deposition, liver function, and liver fibrosis stage.

METHODSNinety-one patients with chronic liver cirrhosis, who underwent megnetic resonance imagine (MRI) scanning in our department between November 2010 and April 2011, were included in the study. A 3.0T MRI scanner was used to acquire T1WI, T2WI, T2WI, and SWI images. The number of nodules, signal intensity ratio (SIR), and contrast noise ratio (CNR) were recorded and analyzed by chi-square and ANOVA statistical tests. Correlation analysis was performed to evaluate the correlations between the number of SN and Child-Pugh classification, ferritin and hyaluronic acid levels.

RESULTSThe sensitivity of SWI, T1WI, T2WI, and T2 WI for detecting SN was 62.5%, 12.1%, 24.2% and 41.8%, respectively. SWI detected significantly more nodules than routine T1WI, T2WI, and T2 WI procedures (P < 0.05). The SIR was the lowest in SWI (0.361 ± 0.209), as compared to T1WI (0.852 ± 0.163), T2WI (0.584 ± 0.172), and T2 WI (0.497 ± 0.196). The CNR was the highest in SWI (13.932 ± 5.637), as compared to T1WI (9.147 ± 5.785), T2WI (9.771 ± 5.490), and T2 WI (11.491 ± 4.573). The correlation coefficients of the number of SN with ferritin, Child-Pugh classification, and hyaluronic acid levels were 0.672, -0.055, and 0.163, respectively.

CONCLUSIONSThe sensitivity and contrast of SWI for detecting SN in patients with liver cirrhosis are higher than conventional MRI. The number of SN can help to assess the degree of iron deposition in patients with liver cirrhosis.

Adult ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Ferritins ; blood ; Humans ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis ; blood ; pathology ; Male ; Middle Aged ; Sensitivity and Specificity

Objective A simple,sensitive and rapid ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method was developed and validated for the determination of ertapenem in human plasma.Methods Using ertapenem-D4 as internal standard,the protein in plasma was precipitated with acetonitrile;chromatographic column:ACQUITY HSS T3(2.1 mm × 50.0 mm,1.8 μm);the mobile phase was 0.1％formic acid aqueous solution(containing 2 mmol·L-1 ammonium formate)-acetonitrile(0.1％formic acid),using a gradient elution;flow rate:0.4 mL·min-1,injection volume:1 μL,column temperature:45 ℃,the analysis time was 4.5 min,the scanning mode is positive ion selective reaction monitoring mode(SRM)with an electric spray ion source(ESI).The specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect,dilution effect and stability were investigated.Results Ertapenem had a good linearity within 0.5-80.0μg·mL-1,and the standard curve was y=4.25 × 10-1x-2.64× 10-2(r2=0.999 0),the lower limit of quantification was 0.5 μg·mL-1,the relative standard deviation within and between batches is 1.39％-4.15％.The extraction recovery rate was 58.36％-64.57％,and the relative standard deviation of dilution effect was 3.30％,and the matrix effect was 99.71％-103.23％.The relative standard deviation of room temperature,repeated freeze-thaw,4 ℃,and long-term stability are all less than 10％.Conclusion The method is sensitive,rapid and specific,which is suitable for clinical monitoring of Ertapenem.

Tooth development relies on sequential and reciprocal interactions between the epithelial and mesenchymal tissues, and it is continuously regulated by a variety of conserved and specific temporal-spatial signalling pathways. It is well known that suspensions of tooth germ cells can form tooth-like structures after losing the positional information provided by the epithelial and mesenchymal tissues. However, the particular stage in which the tooth germ cells start to form tooth-like structures after losing their positional information remains unclear. In this study, we investigated the reassociation of tooth germ cells suspension from different morphological stages during tooth development and the phosphorylation of Smad2/3 in this process. Four tooth morphological stages were designed in this study. The results showed that tooth germ cells formed odontogenic tissue at embryonic day (E) 14.5, which is referred to as the cap stage, and they formed tooth-like structures at E16.5, which is referred to as the early bell stage, and E18.5, which is referred to as the late bell stage. Moreover, the transforming growth factor-β signalling pathway might play a role in this process.