Objective To study the mutation of papillomavirus type 16 (HPV16) E6 gene, and to compare with the result of other researchers. Methods DNA was extracted from cervical cancer tissue sam-pies, then HPV16 E6 genes were amplified and sequenced using the HPV16 E6 specific primers from cervi-cal cancer DNA samples. Results HPV16 E6 gene mutations at the position of nt178 were found in 18 case from 35 cervical cancer samples, the mutation rate was 51.43%, resulting in nucleotide change of Asp to Glu. E6 mutations are found at the position of nt442 in 4 cervical samples. E6 mutation was found in 1 to 2 case in other nucleotide. Conclusion High frequency mutations of E6 gene of HPV16 at the position of nt178, nt442 have been found in cervical cancer samples in Hubei, China. The role of E6 gene mutation in cervical cancer development should be studied further.

Acute Kidney Injury ; chemically induced ; Anti-Infective Agents ; adverse effects ; Child, Preschool ; Humans ; Male ; Organometallic Compounds ; adverse effects

BACKGROUND:Osteoblasts with high purity and activity are essential for bone metabolism research. OBJECTIVE:To explore a simple and effective culturing method of primary osteoblasts. METHODS:Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope;the cells was counted to draw the growth curve;the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining. RESULTS AND CONCLUSION:The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day, and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts.

Animals ; Brain Injuries ; blood ; cerebrospinal fluid ; chemically induced ; Endotoxins ; toxicity ; Female ; Male ; Natriuretic Peptide, C-Type ; analysis ; Neurotensin ; analysis ; Rabbits

OBJECTIVESTo construct the expression vector pGenesil-3-shRNA that can express the short hairpin RNAs (shRNA) silencing Kir6.2 gene and to study its influence on the proliferation and invasion of HepG2 cells.

METHODSTwo shRNA silencing Kir6.2 genes were transcript synthesized intracellularly by expressed templates of plasmid vector pSilence-3, and the target sequence of Kir6.2 gene was inserted into the upstream of the reporter gene in order to construct the recombinant plasmid vector pGenesil-3. Plasmids containing 2 different sequences of human Kir6.2 mRNA coding region were constructed and transfected into HepG2 cells by using lipofectamine 2000 methods. The experiment was divided into 4 groups: SK (normal), SK-HK (negative control), SK-K1 (transfected with the interfering sequence 1) and SK-K2 (transfected with the interfering sequence 2) groups. A selected single clone was cultured after screening by G418. The expression of Kir6.2 protein was detected by Western blot. MTT assay and Transwell system were used to observe the proliferation and invasion of HepG2 cells.

RESULTSThe recombinant expression plasmid pGenesil-3 was successfully constructed and underexpression of Kir6.2 gene in HepG2 cells was detected by Western blot. Underexpression of Kir6.2 gene significantly decreased the proliferation and invasion of the HepG2 cells.

CONCLUSIONshRNA can inhibit the expression of Kir6.2 gene in the HepG2 cells, and underexpression of Kir6.2 gene decreased the proliferation and invasion of the HepG2 cells.

Cell Proliferation ; Genetic Vectors ; Hep G2 Cells ; Humans ; Neoplasm Invasiveness ; Plasmids ; Potassium Channels, Inwardly Rectifying ; genetics ; RNA, Small Interfering ; genetics

Objective To evaluate the stability and biodistribution of a novel SPECT-MRI bi-functional agem 99Tcm-Gd-DTPA-DG in tumor-bearing nude mice. Methods DTPA-DG was synthesized and then conjugated with Gd2O3 to generate Gd-DTPA-DG. The tumor-bearing nude mice were scanned by MRI to evaluate the tumor targeting ability of Gd-DTPA-DG. The orthogonal experiment was applied to optimize pH value of reaction medium and reaction temperature. The radiolabeling efficiency was measured by thin layer chromatography. The distribution of 99Tcm-Gd-DTPA-DG in nude mice was evaluated by scintigrapy in vivo. The % ID/g was measured at different time after intravenous injection of 99Tcm-Gd-DTPA-DG. Results The tumor was significantly enhanced by Gd-DTPA-DG with MRI. The radiochemical purity of 99Tcm-Gd-DTPADG was about 98.5% and remained 96.2% at room temperature for 6 h. The tumor was well visualized by 99TcmGd-DTPA-DG SPECT at 2 h after injection. The tumor uptake was (1.48 ±0.12) %ID/g, and the rumor to muscle radioactivity ratio was 2.91. Conclusions MRI contrast of Gd-DTPA-DG may enhance tumor detection. 99Tcm-labeled Gd-DTPA-DG may be useful for tumor imaging and might have a potential role as a SPECT-MRI bi-functional agent.

OBJECTIVETo find out an effective therapy for obesity induced by antipsychotic agents.

METHODSOne hundred and one cases of obesity who were being treated with antipsychotic agents were randomly divided into an observation group and a control group. The observation group were treated with electroacupuncture at Quchi (LI 11), Zusanli (ST 36), Fenglong (ST 40), Shangjuxu (ST 37) and Xiajuxu (ST 39), once a day for 8 weeks; and no treatment was given to the control group. The Brief Psychiatric Rating Scale (BPRS) was used for assessment of therapeutic effect and the Treatment Emergent Symptom Scale (TESS) for adverse effects.

RESULTSThe clinically effective rate of the observation group (54.9%) was superior to the control group (10.0%) with a significant difference between the two groups (P<0.05). The BPRS score-reducing rate was 24.92% and 28.62% in the both groups respectively, and with no adverse effects.

CONCLUSIONElectroacupuncture has a good therapeutic effect on obesity induced by antipsychotic agents, and it improves the patient's compliance with no adverse effect.

Acupuncture Points ; Antipsychotic Agents ; Electroacupuncture ; Humans ; Obesity

OBJECTIVETo observe the effect of salvia miltiorrhiza and ligustrazine injection on the early myocardial damage of severely burned patients.

METHODSTwenty severely burned patients hospitalized from January 2010 to August 2011, with burn area equal to or more than 50% TBSA, were divided into two groups following hospitalization sequence, with odd number patients entering treatment group (T, n = 10) and even number patients entering control group (C, n = 10). Patients in C group were treated with routine methods, including fluid resuscitation based on the Third Military Medical University formula, anti-infection treatment, support treatment, and organ-protection treatment, etc. In addition to routine treatment methods, patients in T group received intravenous infusion of 250 mL glucose injection (50 g/L) containing 10 mL salvia miltiorrhiza and ligustrazine concoction, once a day, and continued for three days. Venous blood of patients was drawn at post burn hour (PBH) 12, 24, 48, and 72 to determine the plasma levels of cardiac troponin I (cTnI), creatine kinase isozyme MB (CK-MB), and atrial natriuretic peptide (ANP). Data were processed with t test.

RESULTSAt each time point, levels of cTnI, CK-MB, and ANP were lower in T group than in C group. Differences in contents of these parameters between two groups were statistically significant at most time points, with t values from 2.136 to 2.918, P < 0.05 or P < 0.01. Plasma levels of cTnI, CK-MB, and ANP in both groups peaked at PBH 12, which were respectively (28 ± 10) ng/mL, (76 ± 13) U/L, (430 ± 87) pg/mL in T group, and (38 ± 11) ng/mL, (87 ± 10) U/L, (453 ± 91) pg/mL in C group. From PBH 24 to 72, contents of above-mentioned parameters decreased gradually in both groups.

CONCLUSIONSEarly use of salvia miltiorrhiza and ligustrazine injection in severely burned patients can effectively reduce myocardial damage, thus protect the myocardium from injury.

Adolescent ; Adult ; Burns ; blood ; drug therapy ; Creatine Kinase, MB Form ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Pyrazines ; therapeutic use ; Salvia miltiorrhiza ; Troponin I ; blood ; Young Adult

Objective To assess the influence between managements in emergency room(ER) andoutcome of severe traumatic brain injury (TBI),in order to provide inference for treatment.Methods A retrospective analysis was performed in severe TBI patients and recorded next indexes.(1) The managements in ER,including endotracheal intubation and oxygenation,fluid resuscitation,and mannitol intake.(2) The vital signs arriving at ICU,including systolic pressure and blood oxygen saturation.(3) Prognostic indicators including inhospital mortality and days during ICU,the scores of Glasgow outcome scale (GOS) at discharge and 6 months after injury.Results In 140 severe TBI patients,65 patients (46.4％) died during ICU.The mortality of patients with endotracheal intubation [65.0％ (39/60)] was significantly higher than that without endotracheal intubation [32.5％(26/80)](P＜ 0.01).The mortality in whether fluid resuscitation and using mannitol had no significant difference [44.7％ (46/103) vs.51.4％ (19/37),49.2％ (31/63) vs.44.2％ (34/77)] (P ＞0.05).In days during ICU,there was no significant difference among the three treatment measures (P＞ 0.05).In GOS grade at discharge and 6 months after injury,the proportion of 4 and 5 grade were 8.3％ (5/60) and 25.0％ (15/60) in patients with endotracheal intubation,while 27.5％ (22/80) and 52.5％ (42/80) in patients without endotraeheal intubation (P ＜ 0.01).In fluid resuscitation and using mannitol patients,there were no significant difference(P ＞ 0.05).Conclusion Treating severe TBI patients in ER,endotracheal intubation should be carefully chosen,fluid resuscitation and mannitol may not be given.

OBJECTIVETo study the impact of gonadotropin-releasing hormone analogs (GnRHa) on body mass index (BMI) in girls with idiopathic central precocious puberty (ICPP).

METHODSOne hundred and thirty-four girls with ICPP were enrolled. Fifty-seven out of the 134 girls were treated with GnRHa for 1.69±0.43 years. The height, weight, bone age and BMI were measured before treatment, at the end of the treatment and after reaching near adult height and compared with those in the untreated 77 girls.

RESULTSThe adult predicted height standard deviation score (SDS) at the end of treatment was significantly higher than that before treatment (P<0.01) and was similar to the target height SDS in the GnRHa treatment group (P>0.05). With GnRHa treatment, the near-adult height SDS was higher than the target height SDS (P<0.01). At the end of treatment, the BMI SDS slightly increased compared with pretreatment level (P>0.05). A significant reduction in the BMI was observed when reaching the near-adult height in the GnRHa treatment group compared to the level of pretreatment and the untreated group (P<0.01). However, the BMI in the GnRHa treatment group before treatment, at the end of the treatment and after reaching near adult height remained in the normal range (±1 SD).

CONCLUSIONSGnRHa may improve the final height in girls with ICPP. The alterations of BMI after GnRHa therapy fluctuate in a normal range.

Body Height ; drug effects ; Body Mass Index ; Child ; Female ; Follow-Up Studies ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; therapeutic use ; Humans ; Puberty, Precocious ; drug therapy ; physiopathology