Assessment of retrospective dose estimation, with fluorescence in situ hybridization (FISH), of six victims previously exposed to accidental ionizing radiation.

The present study aims to evaluate the use of the fluorescence in situ hybridization (FISH) translocation assay for retrospective dose estimation of acute accidental exposure to radiation in the past. Reciprocal translocation analysis by FISH with three whole-chromosome probes was performed on normal peripheral blood samples. Samples were irradiated with 0-5Gy (60)Co γ-rays in vitro, and dose-effect curves were established. FISH-based translocation analyses for six accident victims were then performed, and biological doses were estimated retrospectively by comparison with the dose-effect curves. Reconstructed doses by FISH were compared with estimated doses obtained by analysis of di-centrics performed soon after exposure, or with dose estimates from tooth-enamel electron paramagnetic resonance (EPR) data obtained at the same time as the FISH analysis. Follow-up FISH analyses for an adolescent victim were performed. Results showed that dose-effect curves established in the present study follow a linear-quadratic model, regardless of the background translocation frequency. Estimated doses according to two dose-effect curves for all six victims were similar. FISH dose estimations of three adult victims exposed to accidental radiation less than a decade prior to analysis (3, 6, or 7 years ago) were consistent with those estimated with tooth-enamel EPR measurements or analyses of di-centrics. Estimated doses of two other adult victims exposed to radiation over a decade prior to analysis (16 or 33 years ago) were underestimated and two to three times lower than the values obtained from analysis of di-centrics or tooth-enamel EPR. Follow-up analyses of the adolescent victim showed that doses estimated by FISH analysis decrease rapidly over time. Therefore, the accuracy of dose estimates by FISH is acceptable only when analysis is performed less than 7 years after exposure. Measurements carried out more than a decade after exposure through FISH analysis resulted in underestimation of the biological doses compared with values obtained through analysis of di-centrics and tooth-enamel EPR.

Inhibition potential of UDP-glucuronosyltransferases (Ugts) 1A isoforms by the analogue of resveratrol, bakuchiol.

Bakuchiol is a promising anti-tumor candidate with resveratrol-like structure. The present study aims to evaluate the inhibition potential of bakuchiol towards UDP-glucuronosyltransferases (UGT) 1A isoforms. An in vitro incubation system using 4-methylumbelliferone (4-MU) glucuronidation was used to evaluate the inhibition capability of bakuchiol towards UGT1A1, 1A3, 1A6, 1A7, 1A8, 1A9 and 1A10. The glucuronidation of trifluoperazine (TFP) was employed as the probe reaction to determine bakuchiol's inhibition towards UGT1A4. At 1 microM and 10 microM of bakuchiol, no or weak inhibition was observed for all the tested UGT1A isoforms. At 100 microM of bakuchiol, the activity of UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9 and 1A10 was inhibited by -46.2%, 74.7%, 17.8%, 98.7%, 70.4%, 99.2%, 75.8%, and 93.3%, respectively. Further inhibition kinetic behaviour was determined for UGT1A6, 1A8, and 1A10. Both Dixon plot and Lineweaver-Burk plot showed the noncompetitive inhibition of bakuchiol towards all these three UGT isoforms. The inhibition kinetic parameters (Ki) were calculated to be 5.3, 1.8, and 92.6 microM for UGT1A6, 1A8, and 1A10, respectively. In combination with the in vivo exposure of bakuchiol, the high possibility of in vivo inhibition of UGT1A6 and 1A8 was predicted. However, relatively low possibility of in vivo inhibition towards UGT1A10 was predicted due to lower in vivo concentration of bakuchiol than its inhibition parameter (Ki). All these information will be helpful for the R&D of bakuchiol as a promising anti-tumor drug.

Cytogenetic analysis in 16-year follow-up study of a mother and fetus exposed in a radiation accident in Xinzhou, China.

In November 1992, a radiation accident occurred in Xinzhou, due to the collection by a farmer of an unused (60)Co source; 37 individuals were exposed to ionizing radiation. Three individuals died and the farmer's 19-weeks-pregnant wife suffered acute radiation symptoms. Conventional chromosome analysis, cytokinesis-block micronuclei (CBMN) assay and fluorescence in situ hybridization (FISH) painting with three pairs of whole chromosome probes were used to analyze chromosomal aberrations for the pregnant female and her baby during the 16 years following the accident. The yields of dicentrics and rings (dic+r) continually declined between 41 days and 16 years after the accident. The frequency of binucleated MN also decreased over time for both mother and daughter. Sixteen years after exposure, the yields of dic+r and binucleated MN decreased to normal levels, but the reciprocal translocation frequencies remained elevated, for both mother and daughter. FISH results showed a decreasing yield of translocations with time. Based on the changes in maternal translocation frequency, the daughter's dose at the time of exposure was estimated as 1.82 (1.35-2.54)Gy. This was consistent with the clinical manifestations of severe mental retardation and low IQ score. FISH-based translocation analysis can be used for follow-up studies on accidental exposure and, after correction, for retrospective dose estimation for individuals prenatally exposed to radiation.

[Correlation study of HPV-16 existential status with Th17/Treg cytokines].

OBJECTIVE: To explore the relationship between the presence of HPV-16 DNA and the expression Treg surface marker Foxp3(+), peripheral blood levels of Th17/Treg cell-associated cytokines and explore their roles and significance in cervical cancer progression. METHODS: Between January 2012 and October 2012 at Shengjing Hospital of China Medical University, a total of 142 HPV16 positive patients were divided into cervical cancer (CC, n = 60), cervical intraepithelial neoplasia (CIN, n = 65) and control group (n = 17). Cervical liquid-based cytological (LBC) samples were collected to detect E2 and E6 genes of HPV type 16 using multiple real-time polymerase chain reaction (PCR). E2/E6 ratio was used to evaluate the physical status of HPV-16 DNA in host cell genome. The SP immunohistochemical method was used to detect the expressions of FOXP3 in cervical lesions. The concentrations of Th17/Treg cell-associated cytokines were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Under the same status of HPV16 DNA in vivo, the levels of Foxp3(+), transforming growth factor-ß (TGF-ß) and interleukin-10 (IL-10) were significantly higher than those of the control group (P < 0.01) while the levels of interleukin-17 (IL-17) and interleukin-21 (IL-21)were significantly lower than those of the control group (P < 0.05) . In the same disease, HPV16 DNA integration rate grew with the increases of Foxp3(+), TGF-ß and IL-10 while IL-17 and IL-21 were opposite. In the different status of HPV16 type DNA, the expression of Foxp3(+) was closely correlated with Federation International of Gynecology and Obstetrics (FIGO) stage, histological grade and lymphnode metastasis (P < 0.05) except for age (P > 0.05). CONCLUSION: Treg cytokines, HPV16 integration rate and severity of cervical lesions are positively correlated while Th17 cytokines show opposite effects. Th17/Treg cell-associated cytokines may play an important role in the occurrence and development of cervical cancer.

[Different methods for the diagnosis of endometrial histological comparative study].

OBJECTIVE: To evaluate the feasibility of endometrial sampler Pipelle for endometrial histologic diagnosis. METHODS: Using prospective and self-control methods, 200 patients with endometrial biopsy firstly used Pipelle endometrial sampler for endometrial tissue, then followed by diagnostic curettage, the same pathologist evaluated the specimen quality and made the histologic diagnosis. RESULTS: Totally 200 patients completed the observation, the specimen satisfaction of Pipelle was 93.0% (in this 200 cases, 186 cases were satisfactory), its pathological accuracy was 85.0% (in this 200 cases, 170 cases' pathological results are highly consistent with diagnostic curettage). There was no significant difference between two kinds of endometrial sampling (P > 0.05). There was no pain for patients during the Pipelle using process. CONCLUSION: Pipelle could obtain satisfactory samples used for histological diagnosis in normal endometrium, simple hyperplasia, complex hyperplasia, atypical hyperplasia and endometrial cancer disease, because its pathological accuracy is so close to the diagnostic curettage, which may be used as a routine screening tool of endometrial diseases.

[Multicenter randomized controlled clinical study for the operative treatment of malignant ovarian germ cell tumors].

OBJECTIVE: To investigate the operative treatment for first-treated patients with malignant ovarian germ cell tumors who need preservation of fertility. METHODS: The clinical data of 105 patients who were treated with fertility-sparing surgery in 11 hospitals from 1992 to 2010 were collected to evaluate the outcomes of different primary surgical operative procedures. All 105 cases were performed the surgeries that preserved fertility and divided into three groups according to the surgical approaches, comprehensive staging surgery group: 47 cases (44.8%) received comprehensive staging surgeries that including the ipsilateral oophorectomy + omentectomy + retropertoneal lymph node dissection ± appendectomy + multiple biopsies;oophorectomy group:45 cases (42.9%)received ipsilateral oophorectomy ± biopsy of contralateral ovary ± omentectomy;tumor resection group:13 cases (12.4%) received enucleation of the mass with preservation of the ovary. Differences were compared among the three groups of patients in the surgery-related indicators, complications, fertility and prognosis. RESULTS: (1) Surgery-related indicators:the average blood loss of the comprehensive staging surgery group, the oophorectomy group and the tumor resection group were 496, 104 and 253 ml, the mean operation time were 176, 114 and 122 minutes, respectively, and there were significant differences among three groups (P = 0.011, P = 0.000). (2) Complication:the surgical complication rates of the three groups were 17% (8/47), 0 and 1/13, with significant differences (P = 0.015). (3) Reproductive function status: the pregnancy rate and birth rate of the three groups were no significant differences (9/19 vs. 7/19 vs. 2/3, P = 0.515; 8/19 vs. 5/19 vs. 2/3, P = 0.636). (4) PROGNOSIS: the recurrence rate of the three groups were significant differences [13% (6/47) vs. 0 vs. 2/13, P = 0.013], but the death rate with no significant differences [6% (3/47) vs. 0 vs. 0, P = 0.129]; The five-year survival rate of three different groups were 89%, 100% and 100% (P > 0.05), while disease free survival rate were 85%, 100% and 83% (P < 0.05), respectively. CONCLUSIONS: Compared with comprehensive staging surgery, oophorectomy group have higher surgical security and satisfactory prognosis, considerable pregnancy rates and birth rate. The tumor resection security may be reliable, but the prognosis is poor.

Clinical observation of peripheral nerve injury in 2 patients with cancer after radiotherapy.

AIM OF THE STUDY: This study aims to analyze the clinical manifestations and sequelae of peripheral nerve radiation damage of two cases of cancer patients after radiotherapy at the corresponding sites in clinical practice and to summarize experiences and lesions in order to provide a reference for future tumor radiotherapy. MATERIAL AND METHODS: Some data of two cases of patients, such as doses of radiotherapy, clinical manifestations and damage occurrence time, were collected and examinations were conducted to define diagnosis. Afterwards, therapies and follow-up were conducted. RESULTS: Case 1 (rectal cancer) was diagnosed as mild left lower extremity nerve damage. After the symptomatic treatment, the disease condition was improved, and there was no tumor recurrence sign. Case 2 (breast cancer) was diagnosed as left brachial plexus damage, and left upper extremity movement function was lost completely. While the analgesic treatment was conducted, anti-tumor relevant treatments were being carried out. CONCLUSIONS: Radiotherapy can cause different extents of radioactive nerve damage. In practice, it is necessary to constantly improve the radiotherapy technology level and actively prevent the occurrence of complications. Once symptoms appear, the diagnosis and treatment should be conducted as early as possible in order to avoid aggravating damage to cause dysfunction and cause lifetime pain to patients.

[Clinical significance of serum squamous cell carcinoma antigen in the diagnosis, treatment and prognosis of cervical squamous cell carcinoma].

OBJECTIVE: To explore the clinical significance of squamous cell carcinoma antigen (SCC-Ag) in the diagnosis, treatment and prognosis of cervical squamous cell carcinoma. METHODS: The serum SCC-Ag concentrations were measured for 1195 patients with cervical squamous cell carcinoma, 54 patients with non-squamous cell type cervical cancer, 325 patients with cervical intraepithelial neoplasm and 69 healthy women treated at Gynecology Ward of Shengjing Hospital, China Medical University from August 2008 to October 2010. And the correlations with their clinical pathological features of squamous cell carcinoma were analyzed and the changes in the diagnosis, treatment and prognosis monitored. RESULTS: Serum SCC-Ag in patients with cervical squamous cell carcinoma showed a sensitivity of 62.32% and a specificity of 90.10% with the optimal cutoff point of diagnosis at 1.45 µg/L. The differences of pretreatment serum SCC-Ag levels were statistically significant in clinical stage, histological differentiation, depth of invasion and lymph node metastasis (P < 0.01). The posttreatment serum SCC-Ag levels of 106 patients undergoing radical surgery and 264 patients on chemotherapy significantly decreased and were significant different with their pretreatment levels (P < 0.05). There was no relationship of serum SCC-Ag levels and human papillomavirus (HPV) opportunistic infection (all P > 0.05). The best threshold values of pretreatment serum SCC-Ag concentration for predicting early postoperative cervical lymph node metastasis and prognosis of cervical cancer were 2.15 and 12.1 µg/L respectively. CONCLUSION: As a relatively specific tumor maker for cervical squamous cell carcinoma, squamous cell carcinoma antigen is correlated with clinicopathological features of cervical squamous cell carcinoma. And it has important clinical reference values in the diagnosis, prognosis, follow-up evaluation and treatment monitoring of cervical squamous cell carcinoma.

[Association of interferon-γ gene polymorphism and risk of cervical HPV infection].

OBJECTIVE: To investigate the association of interferon (IFN) γ gene polymorphisms and risk and prognosis of HPV cervical infection. METHODS: PCR-ASP was used for detecting IFN-γ rs2430561 polymorphism in 179 HPV positive patients and 328 HPV negative normal controls. RESULTS: The frequency of A allele of 63.7% (228/358) was significantly higher than the frequency of T allele of 36.3% (130/358) in HPV positive group (P = 0.045). The frequencies were 41.3% (74/179) in AA genotype and 14.0% (25/179) in TT genotype, women carrying AA genotype increased the risk of HPV infection compare with those with TT genotype (OR = 1.784, 95% CI: 1.031 - 3.088, P = 0.039). During follow-up, the rate of HPV positive again in AA genotype was 83.8% (62/74), while TT genotype was 20.0% (5/25). In the analysis of Kaplan-Meier, the cumulative HPV negative rates of AA, TA and TT genotype exhibited significantly different (P = 0.008). The cumulative HPV negative rate of AA genotype was the lowest (1.1% - 5.9%). CONCLUSIONS: IFN-γ rs2430561 polymorphisms confer the susceptibility to HPV infection. Women with AA genotype exhibited higher risk of infection and inclined to be continuous status and recurrence after HPV infection.

Blockade of T-type Ca(2+) channels inhibits human ovarian cancer cell proliferation.

Regulation of Ca(2+) channels has been implicated in the progression of tumor cells. We report here that T-type Ca(2+) channel expression in human ovarian cancer tissues is greatly increased compared to normal ovarian tissues. Blockade of T-type Ca(2+) channel with NNC 55-0396, mibefradil, or by specifically knocking down the expression of these proteins with siRNA-Ca(v)3.1/3.2 suppressed the proliferation of two ovarian cancer cell lines and increased G0/G1 phase distribution in the cell cycle. Furthermore, NNC 55-0396 slowed ovarian cancer formation in nude mice. Therefore the function of T-type Ca(2+) channels is important for the proliferation of human ovarian cancer cells.

Follow-up study of a pregnant woman 16 years after exposure in the Xinzhou radiation accident.

To investigate the late effects of radiation on child-bearing women, a follow-up study was performed on a 39-year-old survivor 16 years after a (60)Co radiation accident. The woman, Fang, was 19 weeks pregnant at the time of exposure. Physical examinations, a full range of clinical laboratory and imaging tests, as well as cytogenetic analyses were conducted to evaluate Fang's current health conditions. Fang shows the appearance of premature ageing and has a decreased menstrual period. Laboratory studies and imaging tests suggest nodular goitre disease and osteoporosis. Otherwise, no apparent abnormalities were found in the major organs. No malignant tumours were detected by either tumour markers or imaging tests. However, the existence of chromosome aberrations warrants long-term follow-up for tumour incidence in the future. Fang became pregnant 8 years after the accident, but suffered a miscarriage due to the death of the foetus at 6 months into the pregnancy. In conclusion, our findings suggest that the intrauterine death of the foetus might be associated with the previous exposure. There is no evidence of malignant tumours as of the date of the follow-up study. Non-cancerous diseases, i.e. thyroid disease and osteoporosis, which may be related to radiation exposure, are the major manifestations of the long-term effects of the accident.

Higher levels of CCL20 expression on peripheral blood mononuclear cells of chinese patients with inflammatory bowel disease.

This study aimed at characterizing the levels of CCL20 mRNA transcripts in peripheral mononuclear blood cells (PMBC) of 56 Chinese patients with inflammatory bowel disease (IBD), 30 other intestinal diseases and 30 healthy controls by quantitative real time polymerase chain reaction. The levels of CCL20 mRNA transcripts in PBMC of patients with IBD were significantly higher than that of patients with non-IBD intestinal diseases and healthy controls (p < 0.01) and the CCL20 expression in active IBD patients was significantly higher than that in remission patients (p < 0.01). Importantly, the levels of CCL20 expression in PBMC were significantly correlated with the degrees of disease severity, the levels of erythrocyte sedimentation rate and C-reaction protein, but not hemoglobin, in patients with IBD (p < 0.01). Furthermore, the levels of CCL20 expression in active IBD patients after treatment with salazosulphapyridine or prednisone were significantly reduced, as compared with before treatment (p < 0.01). Therefore, analysis of CCL20 expression in PBMC may be used as a surrogate measure for evaluation of IBD activity, disease progression and therapeutic efficacy in Chinese IBD patients.

[Relevant study on apoptosis of cervical cancer HeLa cells induced by paeoniflorin].

OBJECTIVE: To explore the effects of paeoniflorin on the proliferation and apoptosis of HeLa cells. METHODS: HeLa cells treated with paeoniflorin at different concentrations for different hours were assessed by the method of methyl thiazolyl tetrazolium (MTT). Cell apoptosis rate and cycle change were detected by flow cytometry with annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI). The morphological change of HeLa cells was observed by transmission electron microscope (TEM). The expressions of Bcl-2, Bax and Caspase-3 in HeLa cells induced by paeoniflorin were detected by immunocytochemistry. RESULTS: After treatment with different concentrations of paeoniflorin for different hours, the proliferation of HeLa cells was inhibited in a dose and time-dependent manner with an IC50 value of 5054, 2965, 2459 µg/ml at 24, 48 and 72 hours respectively (P < 0.05). After treated by different concentrations of paeoniflorin (control group, 1000 and 2000 µg/ml), apoptosis was induced in HeLa cells at a rate of 0.94%, 10.94% and 13.95% respectively. Such effects were dose-dependent (P < 0.05) and the proportion of HeLa cells in Sphase was under a rising trend. Typical apoptotic changes of HeLa cells under the exposure to paeoniflorin were observed under TEM. There was a lowered expression of Bcl-2 and an elevated expression of Bax and Caspase-3 genes versus the control group after a 48-hour paeoniflorin treatment (P < 0.05). CONCLUSION: Paeoniflorin can significantly induce the apoptosis of HeLa cells through a down-regulation of anti-apoptotic gene Bcl-2 and an up-regulation of pro-apoptotic genes Bax and Caspase-3.

[Surveying on menstruation status of reproductive age women of Han nationality in Liaoning province in 2008].

OBJECTIVE: To investigate the menstruation status of reproductive age women of Han Nationality in Liaoning province. METHODS: From Apr. 2008 to Dec. 2008, 1611 women at age of 19-45 years from Shenyang, Yingkou, Benxi, Zhangwu were enrolled in this study according to epidemiologic cluster sampling method. The study was performed by questionnaire consisting of age of menarche, regularity of menstruation, menstrual cycle and dysmenorrhea, et al. RESULTS: In the survey of 1611 women, the average menarche age is 14.4 years old, there were significant difference on menarche between (14.2 +/- 1.5) years in women from city and (14.6 +/- 1.5) years in women from country (t = 6.58, P < 0.01). The linear regression statistic method was used to analyze the relationship between age and menarche age, the linear regression equation was gotten as Y = 0.074X + 11.855, which means 1 year increase was associated with decrease by 0.074 years in menarche age approximately. About 86.34% (1391/1611) of women have normal and regular menstrual cycle at range of 21 to 35 days, while 11.05% (178/1611) of women have longer menstrual cycle (> 35 days) and 2.61% (42/1611) of women have shorter menstrual cycle (< 21 days). Of which 65.67% (1058/1611) women have regular menstrual cycle just after menarche, 94.97% (1530/1611) of women would have regular menstruation in 2 years after menarche. The rate of dysmenorrhea was 42.09% (678/1611), of which 13.6% (92/678) women have high severe pelvic pain. CONCLUSION: Our results suggested that the trend went toward younger ages of menarche, which was younger menarche age in women from city than country. About 95% women would have regular menstrual cycle within 2 years after menarche.

Semen affects the biological behavior of HeLa cells by altering ERK signaling.

INTRODUCTION: The aim of the study was to investigate the effects of human semen on the proliferation, survival, migration and invasion of HeLa cervical cancer cells by analyzing the extracellular regulated protein kinase (ERK) pathway. MATERIAL AND METHODS: HeLa cells were stimulated with different concentrations of human semen. MTT assay was used to analyze the effects on cell proliferation. Apoptosis in the different experimental groups was quantified by Annexin V-FITC/PI staining. The effect of seminal plasma on in vitro invasiveness of cells was evaluated using transwell assay. Western blotting was used to evaluate ERK pathway activation. RESULTS: Human semen promoted HeLa cell proliferation; ERK1/2 phosphorylation and c-myc expression also increased with increasing semen concentration. U0126 inhibited semen-induced ERK1/2 phosphorylation, c-myc upregulation and cell proliferation. Compared with the control group, semen did not significantly affect the apoptotic rate of HeLa cells (p > 0.05). The transwell assays showed that compared with the control group, the number of invading cells increased significantly with increasing semen concentration, and the difference was significant (p < 0.05) when 1 : 50 semen was added, suggesting that semen promotes the invasion of cervical cancer cells. Western blotting indicated that ERK1/2 phosphorylation began to increase when 1 : 100 semen was added; with increasing semen concentration, ERK1/2 phosphorylation was significantly up-regulated, and the expression of its downstream target gene, c-myc, was also up-regulated (p < 0.05). CONCLUSIONS: Semen promoted the proliferation of HeLa cells by activating the ERK pathway and showed increased tumorigenic potential in vivo. Human seminal plasma might be a potential factor contributing to the development of cervical cancer.

Phase I clinical trial on intracoronary administration of Ad-hHGF treating severe coronary artery disease.

OBJECTIVE: Therapeutic angiogenesis is a new strategy for treatment of vascular insufficiency. Hepatocyte growth factor (HGF)-induced angiogenesis has been applied to induce the neovascularization of ischemic adult tissues in preclinical studies. This report summarizes a phase I clinical trial on the safety of adenovirus-mediated human HGF (Ad-HGF) gene transfer to treat clinically significant coronary artery disease. METHODS: The 18 patients with severe and diffused triple vessel disease determined by coronary angiography, 1-3 of the main coronary arteries not amenable to bypassing grafting and to catheter-based revascularization were assigned to 3 study groups according to the dose of Ad-HGF (from low to high), and the total dose as follows: 5 x 10(9) pfu (group A, n = 6); 1 x 10(10) pfu (group B, n = 6); 2 x 10(10) pfu (group C, n = 6). Arterial gene transfer was performed by over-the wire balloon to the distal of the accessible artery or otherwise the ostium of the target vessels by diagnostic coronary catheter. General safety parameters and cardiac-specific parameters were measured through the preoperative period and on day 7, 21, and 35 postoperatively. The safety and tolerance of Ad-HGF for patients were evaluated according to functional and cytological assessments. RESULTS: During the acute phase up to day 35 and at 11-14 months of follow-up there were no serious adverse events. A mild fever during the first 3 days was not present at day 4, and no long term or paroxysmal fever was found. There were no acute alterations in hemodynamic parameters and the electrocardiogram remained normal. No serious pericardial effusion was reported and there were no arrhythmia on Holter registrations. Moreover, the serum levels of HGF were not changed and the serum anti-adenovirus in the patients was not detected up to day 35. CONCLUSIONS: The present study demonstrates that it is feasible to safely use an adenovirus gene-transfer vector to deliver the human hepatocyte growth factor gene to individuals with clinically significant coronary artery disease by direct intracoronary injection. However, a great deal of additional work must be done before administration of Ad-HGF can be recommended for clinical practice.

Hepatocyte growth factor plays a critical role in the regulation of cytokine production and induction of endothelial progenitor cell mobilization: a pilot gene therapy study in patients with coronary heart disease.

1. There is growing evidence of the beneficial effects of hepatocyte growth factor (HGF) in myocardial infarction, heart failure and occlusive peripheral arterial disease. The aim of the present study was to evaluate the effects of intracoronary administration of an adenovirus vector encoding the human HGF gene (Ad-HGF) on serum levels of cytokines and mobilization of CD34(+) and CD117(+) cells in patients with coronary heart disease. 2. Twenty-one patients with severe coronary artery disease were recruited to the study: 11 patients received both a stent and administration of Ad-HGF; the remaining 10 patients received a stent alone and served as the control group. Blood samples were obtained from the femoral vein before and then 6 and 24 h, 3 and 6 days and 2 weeks after treatment for the isolation of serum and peripheral blood mononuclear cells. Intracoronary administration of Ad-HGF in patients with coronary heart disease resulted in high levels of HGF gene expression, as well as its receptor c-met, compared with the control group, as demonstrated by real-time reverse transcription-polymerase chain reaction. In addition, serum levels of HGF, vascular endothelial growth factor, monocyte chemoattractant protein-1 and interleukin (IL)-10 were increased and serum levels of IL-8 were decreased in patients administered Ad-HGF compared with the control group. The percentage of CD34(+) and CD117(+) cells in the peripheral blood increased in patients administered Ad-HGF. 3. In conclusion, HGF gene therapy may play an important role in the regulation of cytokines and the induction of endothelial progenitor cell mobilization in patients with coronary heart disease.

[Bacterial composition and resistance from urinary tract infections in females].

OBJECTIVE: To determine the bacterial composition and antibacterial resistance in female urinary tract infections (UTI) in China. METHODS: The disc diffusion test, minimum inhibition concentration (MIC), test or E-test was used to detect the antimicrobial susceptibility in 86 tertiary hospitals in China from June 1, 2006 to May 31, 2007. WHONET 5.4 was applied for data analysis. RESULTS: A total of 6071 pathogenic germs were collected from urine cultures of UTI female patients in the surveillance period, which included 5958 bacterial isolates. Escherichia coli 3529 strains (59.23%), Enterococcus spp 938 strains (15.74%) and Klebsiella pneumoniae 394 strains (6. 61%) were the most common pathogens. Escherichia coli and Klebsiella pneumoniae had 35.7% and 34.0% extended spectrum beta-lactamases (ESBL) positive rates, and their resistant rates to quinolones were about 70% and 40%, respectively. Enterococcus faecium was much more resistant to almost all the antibiotics than Enterococcus faecalis. 0.9% Escherichia faecalis and 3. 8% Escherichia faecium were resistant to vancomycin, whereas 2.7% and 4.8% were resistant to teicoplanin. Staphylococcus epdermidis was more common in UTI than Staphylococcus aureus, 79.1% and 75.0% of the isolates being methicillin-ressitant Staphylococcus epdermidis (MRSE) or methicillin-ressitant Staphylococcus aureus (MRSA), respectively. No vancomycin intermediate or resistant staphylococcus was detected, but 4.9% teicoplanin intermediate rate was found in Staphylococcus epdermidis. The resistant rates to quinolones increased with the patient age. CONCLUSIONS: Bacterial resistance in UTI in Chinese females is a severe problem, which could be resulted from antimicrobial irrational use. In the selection of antimicrobial agents in the treatment of UTI, doctors should refer to the results of bacterial resistant surveillance.

[Expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse].

OBJECTIVE: To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP). METHODS: Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade I, 21 patients with grade II, 25 patients with grade III and 12 patients with grade IV in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of I b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5. RESULTS: (1) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2) The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37) in grade III/IV and 26% (8/31) in grade I/IV POP patients, which reached statistical difference (P = 0.035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery < or =2 (19%, 5/27) and >2 patients (12%, 5/41, P > 0.05). (3) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0.002). Among POP group, 19% (7/37) of elastin expression in grade III/IV POP was significantly lower than 45% (14/31) in grade I/II of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60) and non-menopausal patients (3/8), vaginal delivery < or =2 (26%, 7/27) and >2 patients(34%, 14/41, P > 0.05). (4) In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0.031). CONCLUSION: The decreased expression of elastin and fibulin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.

[Effect of Lewis y antigen on regulating gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H].

OBJECTIVE: To investigate the influence of Lewis y antigen on the gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H. METHODS: RT-PCR was used to determine the gene expressions of partial drug resistance associated proteins in RMG-I-H cell line transfected with alpha1, 2-fucosyltransferases gene and RMG-I cell line, as well as in RMG-I-H treated with or without anti-Lewis y monoclonal antibody at the concentration of 10 micro/g/ml. The immunocytochemical method was used to detect the expression of P-glycoprotein (P-gp) in RMG-I and RMG-I-H cell lines. RMG-I and RMG-I-H cells were transplanted into nude mice and the expression of P-gp in the tissues was measured by immunohistochemistry. RESULTS: The mRNA expressions of protein kinase C-alpha (PKC-alpha), topoismerase I ( Topo I ), multidrug resistance-associated protein-1 (MRP-1), and MRP-2 were significantly higher in RMG-I-H cells than those in RMG-I cells (0.46 +/- 0.02 vs. 0.27 +/- 0.05, 0.82 +/- 0.08 vs. 0.52 +/- 0.04, 0.66 +/- 0.07 vs. 0.34 +/- 0.12, and 0.44 +/- 0.08 vs. 0.23 +/- 0.05; all P < 0.05). However, the mRNA expression of multi-drug resistance 1 (MDR-1) was significantly lower in RMG-I-H cells than that in RMG-I cells (0.26 +/- 0.05 vs. 0.45 +/- 0.08, P < 0.05). The P-gp level increased in RMG-I-H cells compared with that in RMG-I cells both in vivo and in vitro (P < 0.05). Expressions of MDR-1, MRP-1, MRP-2, PKC-alpha, and Topo I mRNA decreased by the time in RMG-I-H cells treated with anti-Lewis y monoclonal antibody (all P < 0.05), while mRNA expressions of those genes in the control group did not statistically change (P > 0.05). In addition, MDR-1, MRP-1, MRP-2, PKC-alpha, and Topo I mRNA expressions were significantly lower in RMG-I-H cells treated with anti-Lewis y monoclonal antibody than those in the control group at 6 hours (all P < 0.05) and the inhibition ratios were 48.55%, 77.50%, 70.18%, 45.86%, and 46.13%, respectively. CONCLUSION: The Lewis y antigen of the human ovarian cancer cell surface is closely correlated with the regulation on the gene expression of partial drug resistance associated proteins.