Baculovirus has many advantages as vectors for gene transfer. We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP (Ac-CMV-GFP) could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter. MTT assay suggested that both Ac-CMV-p35 and Ac-CMV-GFP did not have cytotoxic effect on human embryo kidney 293 cells. Cell growth curve showed the Ac-CMV-p35 and Ac- CMV-GFP transduced and non-transduced cells had similar proliferation rate, so baculovirus-mediated p35expression had no adverse effect on cell proliferation. In addition, baculovirus-mediated p35 gene expression protected human embryo kidney 293 cells against apoptosis induced by various apoptosis inducers such as Actinomycin D, UV or serum-free media. These results suggested that the baculovirus vector mediated p35 gene expression was functional and it could be widely used in molecular research and even gene therapy.

Adolescent ; Classical Lissencephalies and Subcortical Band Heterotopias ; diagnosis ; Female ; Humans

The clinical therapeutic effect of acupoint catgut-embedding for simple obesity was systemically analyzed to provide reference and assistance for its clinical treatment and research. By searching in the CBM, CNKI, VIP, Wanfang, Pubmed, Springer and Medline databases, clinical randomized controlled trials (RCT) of acupoint catgut-embedding for simple obesity published from Jan, 2009 to July, 2013 were collected while Revman 5. 2 software was applied to perform the Meta-analysis. Totally 19 articles were acquired with 1 658 cases involved. The effective rate was selected as primary outcome measure in 19 articles. The Meta-analysis was performed among homogeneous researches. The results indicated that compared with other therapies, pooled OR of acupoint catgut-embedding was 2.45 with 95% CI [1.81, 3.32]; in the test for overall effect, Z = 5.81, implying the efficacy difference of two therapies was significant in the treatment of simple obesity (P < 0.01). In subgroups analysis, in the event of treatment session with more than 3 months, compared with other therapies, pooled OR of acupoint catgut-embedding was 2.61 with 95% CI [1.53, 4.46]; in test for overall effect, Z = 3.51, implying the efficacy difference of two therapies was significant in the treatment of simple obesity (P < 0.01); in the event of treatment session with less than 3 months, compared with other therapies, pooled OR of acupoint catgut-embedding was 2.38 with 95% CI [1.65, 3.44]; in test for overall effect, Z = 4.46, implying in the treatment of simple obesity the efficacy difference of two therapies was significant (P < 0.01). Compared with electroacupuncture, OR of acupoint catgut-embedding was 1.79, 95% CI [1.08, 2.95] (P = 0.02). Compared with acupuncture, OR of acupoint catgut-embedding was 1.89, 95% CI [1.16, 3.09] (P = 0.01), which explained that compared with electroacupuncture and acupuncture, the efficacy of acupoint catgut-embedding was significantly different. In a word, the clinical efficacy of acupoint catgut-embedding is superior to other therapies in the treatment of simple obesity.
Acupuncture Points ; Acupuncture Therapy ; Catgut ; Electroacupuncture ; Humans ; Obesity ; therapy ; Randomized Controlled Trials as Topic

Objective To estimate the dietary diversity,overweight and obesity of the rural adults aged 18-65 years in Jilin Province by diet diversity score(DDS),and to analyze the association between dietary diversity and overweight,obesity.Methods A representative sample of 674 rural residents was selected by a multistage sampling method from Jilin Province in 2012 June to July. A validated semi-quantitative food-frequency questionnaire was used to assess the usual food intake. The height and body weight were measured and the body mass index (BMI)was calculated. Logistic regression analysis was applied to calculate the risk of overweight and obesity for different DDS,after adjusted for mixed factors.Results 62.4% people in rural scored ≥6 while 1 1.8% people in rural scored ≤3.The detection rate of obesity of the rural adults in Jilin Province was higher than the mean level in China .For rural adults with moderate and adequate diversity score, the risk of overweight and obesity was 0.946 and 0.816 times the risk of overweight and obesity of the rural adults with pool diversity score. Conclusion Diet diversity of the rural adults in Jilin Province is low.The risk of overweight and obesity is high;the risk of obesity is decreased with the increasing of diet diversity level.

Objective To evaluate the application and initial experience of laparoscopy in kidneypreserving surgery for savage giant hydronephrosis. Methods This series included 6 cases of savage gianthydronephrosis (2 men and 4 women;age range,15 -57 years;mean age,28 years).Of them 5 cases weredetected when visiting doctors due to flank pain,abdominal mass,and the rest one by B-ultrasound duringpregnancy.Four cases had hydronephrosis on the left;and 2 cases,on the right.The quantity of hydronephro-sis was 2250 -8300 ml,respectively.None had development on IVU examination.Of them,3 cases had con-genital ureteropelvic junction (UPJ) obstruction;2 had multiple stones in infracalices secondary to UPJ ob-struction;1 had stones in pelvis with polyp formation.Relieving obstruction,pyeloplasty,nephroplication andnephropexy were performed via laparoscope. Results All the operations were successful.The operativetime was 2.5 -5.0 h;the blood loss was 50 -150 ml,and the mean postoperative hospital stay was 7.2 d.The postoperative follow-up ranged from 3 to 24 months. Three months after operation,B-ultrasound showedthat giant hydronephrosis was markedly relieved in 5 cases (the renal sinus separation was 1.8 m,2.0 cm,2.5 cm,2.5 cm and 2.8cm,respectively),and in the rest 1 case the kidney was slightly smaller than nor-mal.IVU examination was performed every 3 months after operation, and different degrees of developmentappeared in all cases.During the follow-up,no obvious ureteropelvic anastomotic stricture was found on retro-grade pyelography (RGP). Conclusions The protective renal treatment via laparoscopy for savage gianthydronephrosis is a feasible and minimally invasive technique that provides the same clinical and radiograph-ic results as open operation.

To develop a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Norwalk GII. 4 primers which recognized 6 distinct regions on the RNA-dependent RNA polymerase gene of Norwalk GII were designed and used for LAMP assay. Norwalk GII RNA was amplified under isothermal conditions (65 degrees C) for 120 min, and LAMP results were then judged with naked eye, SYBR Green I staining, electrophoretic analysis and restriction digestion. To evaluate the specificity of the RT-LAMP, 48 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses were tested. To compare the sensitivity of the RT-LAMP with that of conventional RT-PCR, Norwalk GII RNA was serially diluted and amplified by RT-LAMP and RT-PCR, respectively. With 46 fecal specimens of Norwalk GII, observation with naked eyes, SYBR Green I staining and electrophoretic analysis were able to detect the PCR products in the RT-LAMP assay. The specificity of RT-LAMP products was also confirmed by digestion of the RT-LAMP products with restriction enzymes. No RNA amplification was observed in 2 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses. The specificity of the RT-LAMP assay with regard to RT-PCR were 100% for Norwalk GII. The detection limits of RT-LAMP was 15.6 pg/tube for Norwalk GII and similar to that of a RT-PCR assay. Compared to RT-PCR, the RT-LAMP assay has been proven to be a rapid, sensitive, specific and accurate method for detection of the Norwalk GII in fecal specimens, and that RT-LAMP assay is potentially useful for the rapid detection of Norwalk GII from fecal specimens in outbreaks of infectious diarrhea.
Caliciviridae Infections ; virology ; Feces ; virology ; Humans ; Norwalk virus ; genetics ; isolation & purification ; RNA Replicase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

OBJECTIVETo observe the effects of Tongfu Decoction (TFD) on the gastric emptying of normal rats, thus exploring whether it could promote gastric emptying rapidly.

METHODSThirty Sprague-Dawley (SD) rats were randomly divided into 3 groups, i.e., the normal control group, the domperidone group, and the TFD group, 10 in each group. They were respectively administered with normal saline, the domperidone suspension, and TFD by gastrogavage. Thirty min later the gastric emptying of mice was detected by single photon emission computed tomography technology (SPECT) labeled with 99m Tc-DTPA, and the gastric half-emptying time and the gastric emptying rate were calculated.

RESULTSThe gastric half-emptying time was (19.0 +/-1.7) min in the normal control group, (12.9 +/- 3.4) min in the domperidone group, and (12.7 +/- 4.1) min in the TFD group. Compared with the normal control group, the gastric half-emptying time was significantly shortened in the domperidone group and the TFD group (P <0.05). The gastric emptying rate at 15 min was 41.1% +/- 5. 8% in the normal control group, 52.9% +/- 10.9% in the domperidone group, and 56.0% +/- 10.3% in the TFD group, while at 30 min it was 65.6% +/- 2.8%, 72.9% +/- 2.6%, and 72.4% +/- 4.9%, respectively. Compared with the normal control group, the gastric emptying rate at 15 min and 30 min both significantly increased in the domperidone group and the TFD group (P <0.05). There was no statistical difference in the gastric half-emptying time or the gastric emptying rate between the two groups (P >0.05).

CONCLUSIONTFD showed similar effects as domperidone in rapidly promoting gastric emptying, and could shorten the gastric half-emptying time in normal rats.

Animals ; Domperidone ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gastric Emptying ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley

Objective Data on the leukapheresis from 26 pediatric patients with hematologic or solid malignancies was retrospectively evaluated to screen predictive factors affecting the efficacy of peripheral blood stem cell(PBSC) collection from donors,as well as hematopoietic recovery in recipients.Methods We present our experience with 49 apheresis from 26 granulocyte-colory Stimulating factor mobilized donors and analyzed the correlations between the mobilization,the leukocyte count in the donor peripheral blood and the MNC and CD_(34)~+ cell yields in collecting products and the neutrophil and platelet recovery of recipients.Results The process of mobilization and apheresis were well tolerated by our pediatric donors.The median numbers for harvested MNCs and CD_(34)~+ cells were 4.5?10~8/kg and 1.9?10~6/kg of recipient body weight,respectively.Mobilizing dose positively affected the number of mononuclear ceus(MNC) but not CD_(34)~+ cells in the apheresis products.The CD_(34)~+ cell number in the apheresis product was influenced significantly by donor circulating MNC on the day of harvest and correlated with recipient′s engraftment after PBSC was reinfused.Conclusions The MNC yield was stable and met with the demand for autologous stem cell transplantation while the CD_(34)~+ cell number varies obviously from each donor.Since a rapid engraftment was associated with a high number of CD_(34)~+ cells collected,which was in turn predicted by the level of the pre-apheresis CD_(34)~+ cells in the peripheral blood of donors,it is necessary to monitor the donors′ CD_(34)~+ cell during mobilization to determine the optimal time for apheresis.J Appl Clin Pediatr,2006,21(3):148-150

OBJECTIVETo explore the clinical value of determination of ATP levels in CD4(+) cells of patients with cytomegaloviral pneumonia after kidney transplantation.

METHODSTwenty-eight patients with cytomegaloviral pneumonia following kidney transplantation and 30 healthy volunteers were enrolled in this study. ATP-bioluminescence assay (ATP-CVA) was used to assess the immune response of CD4(+) cells to phytohemagglutinin (PHA) stimulation in the normal volunteers and the recipients (before and at 1, 2, and 4 weeks after renal transplantation, before and at 2 and 4 week after the treatment).

RESULTSATP concentration in CD4(+) cells of the recipients was 402-/+58 ng/ml before the operation, significantly lower than that in normal volunteers (458-/+196 ng/ml, P<0.05), and reached the lowest level in the first week after operation especially in the recipients with antibody-inducing therapy; ATP level increased slowly since week 2 post-operation, but still remained significantly lower than the preoperative by the fourth week (266-/+87 ng/ml, P<0.05), especially in the recipients receiving antibody-inducing therapy. In the event of cytomegaloviral pneumonia, ATP level underwent a mild reduction to 152-/+78 ng/ml in comparison with the postoperative level at the first week (P>0.05), and was significantly lower than preoperative level (P<0.01); the decrease was especially obvious during the exacerbation of the condition. ATP level then increased slowly after effective treatment, but was still lower than the preoperative level at 4 weeks after the operation (336-/+92 ng/ml, P<0.05).

CONCLUSIONThe determination of ATP level in CD4(+) cells allows more accurate assessment of the cellular immunity in the renal transplant recipients with cytomegaloviral pneumonia to help in the clinical treatment of the patients.

Adenosine Triphosphate ; blood ; Adult ; Aged ; CD4-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Cytomegalovirus Infections ; immunology ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Pneumonia, Viral ; immunology ; metabolism ; virology ; Postoperative Complications ; immunology ; metabolism ; Young Adult

OBJECTIVETo explore the effects of prenatal exposure to low level lead on learning and memory of rat's offspring.

METHODSThe pregnant rats were randomizedly divided into 4 groups and provided with doubly evaporated water in the control group and 125, 250 and 500 mg/L lead acetate solution via drinking water in three exposed groups respectively during the pregnancy. The learning and memory ability of 21-day old and 60-day old offsprings were tested by the Morris water maze and the shuttling and avoiding dark box respectively.

RESULTSThe blood and hippocampus lead concentrations of 1-day old and 21-day old offspring in the 3 lead-exposed groups were significantly increased compared with the control group (P < 0.05), while at the period of 60-day old there was no significant difference between them. The time to find the platform in Morris water maze for 21-day old and 60-day old offsprings of the 3 lead in the exposed groups was significantly increased compared with the control group during the 4 days training (P < 0.05). The times of initiative avoiding in the shuttling and avoiding dark box for 21-day old and 60-day old offsprings in the 3 lead-exposed groups were significantly decreased, while the latency of the initiative avoiding and latency of the passive avoiding were significantly increased compared with the control group (P < 0.05).

CONCLUSIONPrenatal exposure to low level lead impaired the space learning and memory ability of offsprings of the rats, and this kind of influence will continue till the offspring's maturity.

Animals ; Dose-Response Relationship, Drug ; Female ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Maze Learning ; drug effects ; Memory ; drug effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; Random Allocation ; Rats ; Rats, Wistar