Objective To express recombinant Vp1 protein of chlamydiaphage φCPG1, prepare monoclonal antibody against Vp1 protein and utilize it to screen clinical isolates of Chlamydia trachomatis. Methods The Vp1 protein was obtained by prokaryotic expression, and monoclonal antibody against this protein was prepared by hybridoma technique. ELISA and Western blot were used to identify monoclonal antibodies. Then,the monoclonal antibody was prepared in quantity by injecting hybridoma cells into the abdominal cavity of BALB/C mice, and purified by using protein G affinity chromatography. Clinical isolates of Chlamydia trachomatis were screened for the chlamydiaphage by immumofluorescence assay using the prepared monoclonal antibody.Results Purified Vp1 protein was obtained. The monoclonal antibody against Vp1 protein was gained after 3times of sub-clone and consistently identified as IgG1. Three hybridoma cell strains that stably secreted monoclonal antibody were generated. Chromosome analysis of hybridoma cells showed that the mean number of chromosome was 96, most of them were telocentric and a few were submetacentric. The titer of purified monoclonal antibody was more than 1: 12 800. Twenty clinical isolates were screened by using the monoclonal antibody and no positive results were obtained. Conclusions The monoclonal antibody against Vp1 protein of chlamydiaphage φCPG1 is successfully prepared, while no chlamydiaphage is detected by immumofluorescence assay using the prepared antibody in 20 clinical isolates of Ct.

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.

Objective To explore the dose-dependent and time-dependent effect of docetaxel on the expression of mammalian eukaryotic initiation factor 3 subunit A (eIF3a) in lung cancer cell line. Methods The human lung cancer cell line A549 was treated with gradient concentrations of docetaxel for different time. Real-time PCR and Western blot were used to detect mRNA and protein expression levels of eIF3a and α-tubulin, respectively. Results Docetaxel did not affect α-tubulin expression at either mRNA level or protein level. When A549 cells were treated with high concentration of docetaxel (30 μg/L), the expression level of eIF3a mRNA tended to increase in a time-dependent manner. Protein expression level of α-tubulin was not associated with eIF3a expression significantly in cells treated by docetaxel.Conclusion Docetaxel could slightly increase the expression of eIF3a mRNA, and eIF3a does not regulate the expression of α-tubulin in A549 cells treated by docetaxel.

Objective To understand the status of drinking-water supply and the progress of waterimproving projects in drinking-water-borne endemic fluorosis areas in Henan,and to provide scientific data for related government departments to carry out prevention and surveillance plan for those areas.Methods Questionnaire survey was carried out in all the villages in drinking-water-borne endemic fluorosis areas in Henan in 2010.Each village was given one set of questionnaire.Results By the end of 2010,the total number of fluorosis villages in Henan was 25 434,among them 11 484 villages had been conducted water-improving projects,accounting for 45.15％.Currently,9267 water-improving projects worked properly,accounting for 36.4％(9267/25 434)of all villages surveyed,and 80.7％ (9267/11 484)villages had water-improving projects.The projects in 2217 villages do not work properly,accounting for 19.3％(2217/11 484) of all projects.There were 5832 water-imp.roving projects conducted amnog 2005-2010,accounting for 50.8％ (5832/11 484) of all water-improving projects.Among villages with no such projects,97.5％(15 769/16 167) of them used shallow groundwater,and in villages with water-improving projects,89.6％ (8303/9267) of them used deep groundwater as drinking-water supply.Conclusions After 2005,the water improving progress was significantly speed up in drinking-water-borne endemic fluorosis areas in Henan,but progress of the water improving projects is relatively slow due to changing and expanding of fluorosis areas.The government should increase investment and improve the proportion of waterimproving defluoridation.

Objective To summarize the surgical experience in robotic-assisted laparoscopic partial nephrectomy,and to investigate the efficacy and safety of this surgery.Methods The clinical data of 12 patients who underwent robot-assisted laparoscopic partial nephrectomy in Changhai Hospital from March to July in 2012 were analyzed.All the patients were male and the age range was 43-66 years.In 4 cases the tumors were in the left kidney,and 8 in the right.In 7 cases the tumors were in the dorsal part of the kidney,and 2 in the ventral part.There were 3,5 and 4 cases in the upper,middle and lower pole of the kidney respectively.Preoperative GFR test was normal in all cases.Kidney CT scan showed the maximum diameters of the tumors were 2.0-5.8 cm,with an average of 3.3 cm.The pre-operative stages in all cases were T1N0M0.Results The surgery was successfully completed in all cases.The mean duration of the surgery was 160-310 min,with an average of 242 min.The blood loss was 30-300 ml,with an average of 135 ml,and the intraoperative blood transfusion was unnecessary.The warm ischemia time was 20-49 min,with an average of 31 min.There was no intraoperative morbidity,and no conversion to open surgery.The postoperative length of hospitalization was 9-31 d,with an average of 14 d.Gross hematuria arose in 1 patient at 1 week after the surgery.The post-operative pathology showed renal clear cell carcinoma with Furhman Grade Ⅱ in 11 cases,and renal angiomyolipoma in 1 case.The maximum diameters of the tumors were 2.0-5.0 cm,with an average of 3.5 cm.The tumor resection margin was negative in all cases.Conclusions Robot-assisted laparoscopic partial nephrectomy is safe and effective for local renal tumors.This surgery has significant advantage over traditional laparoscopic partial nephrectomy,in terms of the resection of the renal tumors and the reconstruction of the kidney.

OBJECTIVETo investigate the suppressive effect and its frequency selection of dopamine on the cochlear auditory afferent nerve activity. To offer an important step in understanding the modulation of dopamine in the inner cell synaptic complex.

METHODSForty guinea pigs were randomly divided into four groups and the whole intracochlear perfusions were performed: (1) perfused with artificial perilymph solutions; (2) perfused with artificial perilymph solutions containing 10 mmol/L dopamine; (3) perfused with artificial perilymph solutions containing 30 mmol/L dopamine; (4) perfused with artificial perilymph solutions containing 50 mmol/L dopamine. Compound action potential (CAP) evoked by different frequencies (250 Hz, 500 Hz, 1000 Hz, 2000 Hz, 4000 Hz, 8000 Hz, 16 000 Hz) and cochlear microphonics (CM) evoked by 4000 Hz tone burst were recorded from the round window of guinea pigs before perfusion and 1 hours, 2 hours after perfusions.

RESULTSThere was no significant difference in CAP threshold before and after perfusion in the artificial perilymph solutions group (P > 0.05). An increase of CAP threshold of most detecting frequencies were observed in the three dopamine-perfused groups (P < 0.05). The inhibition effect intended with the increasing of the concentration of dopamine in the perfusion solution. There was significant difference in CAP threshold shift between different frequencies, especially in the group perfused with 30 mmol/L dopamine. The maximal threshold shift was found at 4000 Hz and 8000 Hz. No significant changes of CM amplitude and non-linearity of input-output function were observed before and after perfusion with artificial perilymph solutions and dopamine.

CONCLUSIONSDopamine can inhibit the cochlear auditory afferent nerve, but there is no obvious influence on outer hair cell produced by dopamine. The inhibition effect has frequency selection, the suppression on high frequencies is more stronger than low frequencies.

Animals ; Cochlear Nerve ; drug effects ; physiology ; Dopamine ; pharmacology ; Evoked Potentials, Auditory ; drug effects ; Female ; Guinea Pigs ; Male

Objective To investigate the water quality and running status of the dringking water improving project in Xuchang,Pingdingshan and Nanyang,thus to provide basis for scientifically evaluating social effects.Methods Stratified sampling was used in the levels of counties and townships and villages,with the number of checked counties greater than or eaquel to half of the projected counties(13/24),the checked townships more than 40 percent of the projected townships(42/102),the checked villages greater than or eaquel to 30% of the projected villages(56/186).We listened their statement,reviewed the documents,examined carefully the water projects before we made the appraisal.Results More than 95%(129/136)of the projects had water quality reports,among which 90%(122/136)were provided by county level centers for disease prevention and control,75% (102/136) of water samples were collected by centers for disease prevention and control,80%(109/136)ofcounties organized an acceptance check-up group,92%(125/136)projects well preserved water source.Conclusions The dringking water improvement projeets in countryside are basically normal in terms of construction and management,the water supplied is qualified,so the expected goal is achieved.However,duties of each department are not explicit and the communication is inadequate,so collaboration should be reinforced.

OBJECTIVE: To investigate the clinical significance of urinary epidermal growth factor (EGF) in patients with brain tumors. METHODS: The levels of EGF in urine samples collected from 20 patients (9 low grade astrocytomas, 6 anaplastic astrocytomas, and 5 meningiomas) and 5 healthy individuals were determined. EGF levels were measured by radioimmunoassay technique. A preoperative and one postoperative determination were performed. RESULTS: Preoperative urinary EGF levels of astrocytoma patients were statistically higher than those of meningioma patients and the controls (P < 0.01). Preoperative urinary EGF levels showed a positive correlation with the degree of malignance in the astrocytoma patients (P < 0.05). A significant decrease of the postoperative levels of EGF was observed in the astrocytoma patients who underwent gross total resection (P < 0.01). The pre/postoperative urinary EGF levels of the meningioma patients showed no significant fluctuations and showed no significant difference with those of healthy individuals (P > 0.05). CONCLUSION The urinary EGF levels of astrocytoma patients correlate with the WHO grade of malignance and significantly decrease after gross total removal. Urinary EGF may be of practical value in diagnosing and evaluating the surgical efficacy of astrocytomas.
Adolescent ; Adult ; Aged ; Astrocytoma ; urine ; Biomarkers, Tumor ; urine ; Brain Neoplasms ; urine ; Epidermal Growth Factor ; urine ; Female ; Humans ; Male ; Meningioma ; urine ; Middle Aged

Objective:To retrospectively analyze the data of patients undergoing extracorporeal membrane oxygenation (ECMO) during perioperative period of cardiac transplantation and provide objective and reliable evidence for further clinical promotion.Methods:Collect the clinical data of patients undergoing heart transplantation and ECMO support in Fuwai Hospital, analyze the duration of ECMO support, combined use of aortic balloon counterpulsation (IABP), and complications during the supporting period. All statistical analyses were processed by SPSS 23.0 software. Independent sample Student's test was employed for normal distribution and Mann-Whitney U test for abormal distribution. χ2 or Fisher exact test was utilized for comparinge the classification data between groups. Results:All ECMO support models were intravenous-arterial ECMO (V-A ECMO). Eight patients successfully bridging heart transplantation through VA-ECMO. Sixty-one patients (89.7%) who had undergone cardiac transplantation were successfully weaned from ECMO while 48 patients (70.5%) survived and discharged. The most common complications during circulation support are bleeding, acute renal insufficiency, and pulmonary infection. Patients with ECMO support in the operating room had a better rate of survival and weaning off(95.6%, 84.4%) than those with ECMO at the bedside of ICU(72.2%, 27.8%).Conclusions:ECMO can provide adequate circulation and respiratory function support for heart transplant patients, and it is an indispensable treatment for patients to go through the perioperative period of heart transplant surgery smoothly. It is advocated to use IABP combined with ECMO in the early stage and at the same time to increase the perfusion of vital organs, improve the prognosis of patients and obtain good outcomes.

OBJECTIVETo discuss diagnosis and treatment of iatrogenic purulent lumbar spinal infection.

METHODSFrom December 2006 to January 2010, 4 patients with iatrogenic purulent lumbar spinal infection were treated with posterior debridement. There were 2 males and 2 females, ranging in age from 50 to 66 years (respectively in 52, 66, 58, 50 years); in course of disease from 2 weeks to 2.5 months (respectively in 21, 14, 60, 75 days ). All patients had fever, lumbago, local tenderness and limited lumbar activity before operation. White blood cell count (WBC), erythrocyte sedimentation rate (ESR) were abnormal. The clinical effects were evaluated by symptoms and laboratory examination.

RESULTSSymptoms of lumbago and fever vanished in 4 patients, of which wounds were primary healing without complications. The patients were followed up for 3 months, no infection (WBC, C-reactive protein and ESR were normal) and lumbar instability were found.

CONCLUSIONIatrogenic purulent lumbar spinal infection can be diagnosed according to course of disease, clinical symptoms and signs, imaging finding. In the items, magnetic resonance imaging finding have necessarily specificity, once finding abscess-formation, will promptly operate.

Aged ; Debridement ; methods ; Diagnosis, Differential ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spondylitis ; diagnosis ; surgery ; Suppuration