Objective To investigate the expression and correlation of hypoxia-inducible factor-1?(HIF-1?),vascular endothelial growth factor(VEGF)and sFlt-1 in the preeclampsia placenta,and discuss their significance in the pathogenesis of preeclampsia.Methods Placentas were collected from 20 pregnant women with preeclampsia as study group and 15 normal pregnant women as control group.The expressions of HIF-1?,VEGF and sFlt-1 protein were semi-quantitatively analyzed with immunohistochemical assay and mRNA level was determined using reverse transcription polymerasc chain reaction(RT-PCR)technique. Results(1)the expression of HIF-1?and sFlt-1 protein in preeclampsia group obviously increased.Strong (+++)positive expression was observed in 9 and 11 cases respectively,significantly higher than in control group(2 and 3 cases)(P＜0.05),however,VEGF expression obviously reduced in preeclampsia group(P＜0.01).(2)the level of HIF-1?and sFlt-1 mRNA in preeclamptic placenta was 0.604?0.013, 0.898?0.041,significantly higher than 0.208?0.007 and 0.559?0.244 in normal placenta(P＜0.05). Although the level of VEGF mRNA increased in preeclampsia placenta,it was not significantly different from that in normal placenta(P＞0.05).The ratio of VEGF mRNA/sFlt-1 mRNA obviously reduced in preeclampsia group and was significantly lower than in control group(P＜0.05).(3)in preeclampsia group,HIF-1?mRNA expression was positively correlated with the expression of sFlt-1 mRNA(r=0.577, P＜0.05),and negatively correlated with the ratio of VEGF mRNA/sFlt-1 mRNA(r=-0.376,P＜0.05).Conclusion Abnormal high HIF-1?expression in preeclampsia placenta indicates that HIF-1?might play an important role in the pathogenesis of preeclampsia,possibly through affecting the cytotrophoblastic invasion and placental vascular reconstruction via the modulation of VEGF and sFlt-1 gene transcription.

Organic solvent tolerant microorganism(OSTM) is a novel extremophile and it hasn't been systematically studied until 1980s.Relying on certain mechanisms,the OSTM is able to effectively de-fend and decrease the toxicity from organic solvents,which enable the OSTM to be potentially applied in the industrial fields such as whole-cell catalysis and environmental treatment,etc.The comprehen-sively understanding of the mechanisms involved in organic solvent tolerance of OSTM could be com-bined with genetic engineering in order to modify and optimize the various specifications of OSTM,and further broaden its application in other industrial areas.Latest studies on the tolerant mechanisms of OSTM,in this paper,will be reviewed from four aspects such as vesicle exocytosis and changes of phos-pholipid composition in membrane,etc.Besides,the application of OSTM in whole-cell catalysis and other fields will be introduced.

To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.

OBJECTIVETo evaluate clinical effect and safety of floating needle therapy and duloxetine in treating patients with persistent somatoform pain disorder (PSPD).

METHODSTotally 108 PSPD patients were randomly assigned to the floating needle treatment group, the duloxetine treatment group, and the placebo treatment group, 36 in each group. Patients in the floating needle treatment group received floating needle therapy and placebo. Those in the duloxetine treatment group received duloxetine and simulated floating needle therapy. Those in the placebo treatment group received the placebo and simulated floating needle therapy. All treatment lasted for six weeks. Efficacy and adverse reactions were evaluated using Simple McGill pain scale (SF-MPQ) and Treatment Emergent Symptom Scale (TESS) before treatment and immediately after treatment, as well as at the end of 1st, 2nd, 4th, and 6th week of treatment, respectively. Hamilton Depression Scale (HAMD, 17 items), Hamilton Anxiety Scale (HAMA) were assessed before treatment and at the end of 1st, 2nd, 4th, and 6th week of treatment, respectively. Patients in the floating needle treatment group and the duloxetine treatment group with the total reducing score rate of SF-MPQ in Pain Rating index (PRI) ≥ 50% after 6 weeks' treatment were involved in the follow-up study.

RESULTS(1) Compared with the same group before treatment, SF-MPQ score, HAMD score and HAMA total scores all decreased in all the three groups at the end of 1st, 2nd, 4th, and 6th week of treatment (P < 0.05, P < 0.01). Besides , each item of SF-MPQ significantly decreased immediately after treatment in the floating needle treatment group (P < 0.01). Compared with the placebo treatment group, SF-MPQ, HAMD, and HAMA total score in the floating needle treatment group significantly decreased after 1, 2, 4, and 6 weeks of treatment (P < 0.05, P < 0.01). SF-MPQ score, HAMD score and HAMA total score in the duloxetine treatment group also significantly decreased after 2, 4, and 6 weeks of treatment (P < 0.05, P < 0.01). (2) There were 3 patients (8.3%) who had adverse reactions in the floating needle treatment group, 17 (50.0%) in the duloxetine treatment group, and 7 (21.2%) in the placebo treatment group. Compared with the placebo treatment group, the incidence of adverse reaction increased in the duloxetine treatment group (χ² = 6.04, P < 0.05). Besides, it was higher in the duloxetine treatment group than in the floating needle treatment group (χ² = 14.9, P < 0.05). (3) There were 19 patients in the floating needle treatment group and 17 patients in the duloxetine treatment group involved in the follow-up study. Compared with 6 weeks after treatment, no significant difference was observed at 3 and 6 months after treatment in the score of SF-MPQ, HAMD, and HAMA in the floating needle treatment group and the duloxetine treatment group. No significant difference was observed between the two groups (P > 0.05). There were 5 patients (29.4%) who had adverse reactions in the duloxetine treatment group, and no adverse reactions were observed in the floating needle treatment group. The adverse reaction rate was significantly different between the two groups (χ² = 4.26, P < 0.05).

CONCLUSIONSFloating needle therapy and duloxetine were effective in treatment of patients with PSPD. However, floating needle therapy could relieve pain more rapidly than duloxetine, with obviously less adverse reactions.

Acupuncture Therapy ; methods ; Analgesics ; therapeutic use ; Anxiety Disorders ; Duloxetine Hydrochloride ; therapeutic use ; Follow-Up Studies ; Humans ; Needles ; Pain ; Pain Management ; methods ; Pain Measurement ; Psychiatric Status Rating Scales ; Somatoform Disorders ; therapy ; Treatment Outcome

OBJECTIVETo study the antitumor effect of saponin extracted from Tupistra chinensis Baker (STCB) against mouse sarcoma S-180 cell proliferation in vitro and in vivo and explore the primary mechanism of this effect.

METHODSCytotoxic effect of STCB on S-180 cells in vitro was evaluated by MTT colorimetry, and its effect against in vitro tumor growth was tested in Kunmin mice bearing S-180 implanted tumor. The morphological and ultrastructural changes of S-180 cells after saponin treatment in vitro were examined with light and transmission electron microscope. Flow cytometry was performed to examine the cell cycle and apoptosis of S180 cells treated with different concentrations of STCB with propidium iodide staining.

RESULTSSTCB could markedly inhibit S-180 cell proliferation in vitro with 50% inhibitory concentration of 34.64 microg/ml. STCB given by intragastric administration also significantly inhibited the growth of S-180 solid tumor, and the inhibition rate exceeded 30% at the dose of 0.5 g/kg, reaching 54.86% at 2 g/kg. Electron microscopy and flow cytometry revealed increased S180 tumor cell apoptotic rate with the increment of saponin concentration, along with increased percentage of cells in S phase and decreased cells in G(2)/M phase in response to 10 or 30 microg/ml STCB treatment. At the concentration of 60 microg/ml, however, STCB resulted in an opposite effect on the cell cycles, presumably due to its interference with mitosis at high concentrations.

CONCLUSIONSSTCB inhibits the growth of S-180 cells both in vivo and in vitro possibly by inducing cell apoptosis and interfering with the cell cycle progression of the tumor cells.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Liliaceae ; chemistry ; Male ; Mice ; Phytotherapy ; Saponins ; pharmacology ; therapeutic use ; Sarcoma 180 ; drug therapy ; pathology

OBJECTIVETo determine if Ganoderma polysaccharides can antagonize prostaglandin E2 (PGE2)-induced suppression of murine splenocyte interferongamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) mRNA expression.

METHODSMixed lymphocyte culture reaction was used as the experimental model. The expressions levels of IFN-gamma and TNF-alpha mRNA were measured by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSAfter the cultures were treated with PGE2 for 4 h, IFN-gamma mRNA expression was reduced as compared with the control, which was especially obvious when PGE2 concentrations exceeded 10 micromol/L (P<0.01). Ganoderma polysaccharides above 100 mg/L showed partial antagonistic effect against the inhibition of IFN-gamma by PGE2 at the fixed concentration of 20 micromol/L. Further studies indicated that PGE2 (20 micromol/L) impaired the expression of TNF-alpha mRNA after an 8-hour incubation and Ganoderma polysaccharides above 100 mg/L could partially antagonize this effect.

CONCLUSIONGanoderma polysaccharides can partially antagonize PGE2-induced suppression of murine splenocyte IFN-gamma and TNF-alpha mRNA expression.

Animals ; Cells, Cultured ; Dinoprostone ; pharmacology ; Female ; Gene Expression ; drug effects ; Interferon-gamma ; genetics ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Polysaccharides ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Reishi ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; cytology ; Tumor Necrosis Factor-alpha ; genetics

Translational medicine is a systemic project because it is patient and clinical problems oriented, aiming at research results application, and involves multidisciplinary cooperation. Studies on molecular events in the precancerous stage, early stage and metastasis of colorectal cancer (CRC) are the CRC hot research topics currently. Investigations on the earliest molecular events can help to find out the markers which may improve the effect of CRC screening and predict CRC liver metastasis and prognosis. Based on the concept of micro environment, molecular targeted drugs to interfere with metastasis and invasion and new concepts of surgical resection margin and neoadjuvant therapy will gain recognition from clinicians.
Colorectal Neoplasms ; Humans ; Translational Medical Research

OBJECTIVETo study the clonality of gastric carcinoma and precancerous lesions and its relationship with Ki-67 protein expression.

METHODSFormalin-fixed paraffin embedded tissues were collected from 174 cases of gastric endoscopic biopsies and surgical removed specimens. The lesional tissues were isolated by Laser Capture Microdissection. Methylation sensitive restriction enzyme (HpaII) digestion and polymerase chain reaction (PCR) were used to detect the clonality at the polymorphic human androgen receptor gene locus on the X chromosome. PCR products were analyzed by capillary electrophoresis using applied Biosystems 3730 DNA Analyzer. In addition, a two-step immunohistochemical staining EnVision method was used to detect the expression of Ki-67 protein.

RESULTSThe frequency of detection of monoclonality and expression rate of Ki-67 were found increased in a stepwise fashion from gastrointestinal metaplasia, low grade intraepithelial neoplasia, high grade intraepithelial neoplasia to intestinal carcinoma (15.63%, 5/32; 22.22%, 10/45; 69.44%, 25/36 and 100.0%, 20/20; respectively). The presence of clonal proliferation was correlated with Ki-67 expression in low grade intraepithelial neoplasia (P < 0.01).

CONCLUSIONSThe presence of clonal proliferation and increased Ki-67 are increasingly detected in the lesions along the multi-step gastric carcinogenesis model. Clonal status is associated with the expression rate of Ki-67 to a certain extent, suggesting a combined application of both markers may be useful in assessing early stages of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Cell Line, Tumor ; Female ; Humans ; Ki-67 Antigen ; genetics ; metabolism ; Middle Aged ; Neoplasm Staging ; Precancerous Conditions ; genetics ; metabolism ; pathology ; Stomach Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo investigate the immunological mechanism of allergic dermatitis induced by trichloroethylene (TCE).

METHODSThe guinea pig model of TCE-induced allergic dermatitis was established by Guinea pig Maximization Test. The effects of TCE and its metabolites on splenic lymphocytes of TCE-sensitized and non-sensitized guinea pig were detected by MTT assay.

RESULTSFor TCE-sensitized guinea pig, the survival rate of lymphocytes cultured with TCE (+S9) was significantly higher than that cultured with TCE (-S9) (83.0% +/- 3.4% vs 75.9% +/- 7.9%, P < 0.01), while, for normal animals, the survival rate of lymphocytes cultured with TCE (+S9) was significantly lower than that cultured with TCE (-S9) (63.4% +/- 8.4% vs 77.0% +/- 7.2%, P < 0.01). The survival rate of lymphocytes cultured with TCE (+S9) in TCE-sensitized animals was higher than that in normal animals (83.0% +/- 3.4% vs 63.4% +/- 8.4%, P < 0.05), but no statistically significant difference was found for TCE (-S9) (75.9% +/- 7.9% vs 77.0% +/- 7.2%, P > 0.05).

CONCLUSIONCytotoxicity of TCE to normal lymphocytes and proliferation of sensitized lymphocytes were enhanced by metabolic activation. The metabolites of TCE may act as effective immune hapten to stimulate the proliferation of hapten-specific lymphocytes in TCE-sensitized animals.

Animals ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dermatitis, Allergic Contact ; etiology ; immunology ; Female ; Guinea Pigs ; Lymphocytes ; drug effects ; Male ; Passive Cutaneous Anaphylaxis ; drug effects ; Spleen ; drug effects ; immunology ; Trichloroethylene ; toxicity

OBJECTIVETo establish a simple, rapid and economical method in detecting mutations of oncogene K-ras and to investigate its mutations in colorectal cancer tissues and its relationship with clinicopathologic characteristics of colorectal carcinoma.

METHODSForty colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using polymerase chain reaction (PCR) followed by direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by sequence analysis. The other 113 colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using PCR-RFLP followed by sequence analysis only. The mutation results were analyzed with the corresponding clinical pathological data.

RESULTSAmong 40 colorectal cancer cases, none of K-ras mutations at codon 12 and codon 13 was detected by PCR followed by direct sequencing. However, K-ras mutations were found in 11 cases (11/40, 27.5%) by PCR-RFLP followed by sequence analysis, including 8 cases at codon 12 and 3 cases at codon 13 respectively. Among 153 colorectal cancer cases, point mutations were detected by PCR-RFLP followed by sequence analysis in 58 cases (37.9%). Point mutations at codon 12 were found in 46 cases and 12 cases at codon 13. Mutations with the highest frequency were G→A transitions (25/58, 43.1%) at codon 12. No significant correlation was observed between mutations of K-ras and gender, invasive depth, tumor differentiation, number of invaded lymph nodes, distant metastasis and clinical stage (P > 0.05). Mutation of oncogene K-ras at codon 12 and codon 13 was closely related with age and tumor location (P < 0.05). The incidence of K-ras mutation was significantly higher in younger patients and in patients with ascending colon cancer.

CONCLUSIONSPCR-RFLP followed by sequence analysis is a rapid, simple, sensitive and low-cost method. It is a suitable technology for detecting hot-spot mutations in the K-ras oncogene. Mutation of oncogene K-ras at codon 12 and codon 13 is a common molecular event in colorectal carcinogenesis, which might be related with age and tumor location.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Polymorphism, Restriction Fragment Length