Adolescent ; Child ; Child, Preschool ; Electroencephalography ; Female ; Humans ; Infant ; Retrospective Studies ; Rett Syndrome ; pathology ; physiopathology

This study is to investigate the antitumor activity of CIP-36 on multidrug resistant human oral squamous carcinoma cell line (KBV200 cells) in vitro and the possible anticancer mechanisms. MTT assay, Hoechst fluorescein stain, RT-PCR and immunohistochemistry were carried out on KBV200 and KB cells. The growth of many tumor cells was obviously inhibited by CIP-36, especially the multidrug resistant cells KBV200. Obvious apoptosis could be observed in the Hoechst 33342 staining experiments. The results of RT-PCR showed that the levels of p53, p21, caspase-3 and bax mRNA increased, and meanwhile the expression of mdr-1 and bcl-2 mRNA decreased in a dose-dependent manner. The data were significantly different from that of vehicle. The expression of P-gp significantly decreased with the increasing dosage of CIP-36 examined by immunohistochemistry. It can be concluded that CIP-36 could change resistance-related genes and proteins to overcome multidrug resistance in the KBV200 cell line.

Objective To explore the effect of pedicle fixation for treatment of thoracolumbar burst fractures in patients with osteoporosis,and to provide more evidence for the treatment.Methods Retrospectively analyzed the clinical data of 121 patients with osteoporotic vertebral burst fracture from June 2012 to October 2015.And these patients were divided into two groups according to different operation methods, namely the control group (n=56) who were given short segment fixation and the observation group (n=65) who were given single segment fixation.The visual analogue scale(VAS),Oswestry disability index(ODI),vertebral height,kyphotic angle and bone mineral density of the two groups were analyzed before surgery and 3 days,1 month,3 months and 12 months after surgery.Results The VAS score,ODI score,vertebral height,and Cobb angle of the injured vertebra were significantly improved in both of the two groups,and the difference was statistically significant (P<0.05).The VAS score of the observation group was better than that of the control group 3 days after surgery with statistically significant difference (P<0.05).But there was no significant difference 3 months,6 months and 12 months after surgery(P>0.05).The ODI score of the observation group was better than that of the control group 3 days and 3 months after surgery with statistically significant difference (P<0.05),and there was no significant difference between the two groups till the end of follow-up.Pedicle fixation at the injured vertebra significantly improved the vertebral height and Cobb angle with statistically significant difference (P<0.05).And the anti-osteoporosis treatment significantly increased the bone mineral density (P<0.05).Conclusion Pedicle fixation at the injured vertebra is useful in pain relief as well as function and anatomical structure restoring.And anti-osteoporosis treatment is necessary for the bone mineral density increase.

Objective To investigate the effects of Epigallocatechin gallate(EGCG)on IL-1βinduced MIN6 cells apoptosis. M.Methods The experiment group was divided into control group,IL-1β group,IL-1β+ EGCG low concentration group and IL-1β+EGCG high concentration group.Cell activity was detected by CCK8.Insulin secretion was detected by ELISA.cell apoptosis was detected by flow cytometry.The mitochondrial membrane potential was detected by flow cytometry.ATP content and cell ac-tivity of ROS were detected by colorimetry and chemiluminescence method.Results Compared with normal group,IL-1β group showed much lower cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,and at the same time IL-1βgroup had significantly higher cell apoptosis and ROS activities.After given EGCG,both low concentration group and high con-centration group had higher cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,at the same time lower cell apoptosis and ROS activities was showed.And the IL-1β+EGCG high concentration group worked more powerful.Con-clusion EGCG has protective effects on IL-1βinduced MIN6 cells apoptosis.Its mechanism may be related to increasing the content of the ATP and mitochondrial membrane potential and protecting mitochondrial function as well reducing the activity of ROS.

A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals.Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits,Ceditest(R)FMDV-NS (Ceditest(R) kit),UBI(R) FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI(R) kit) and a FMDV 3ABC-I-ELISA kitdeveloped at the Lanzhou Veterinary Research Institute.The test parameters (sensitivity and specificity) of the three kits were determined,and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits.The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest(R) kits was 98.05%,and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI(R) kits was 94.4%; the sensitivity of both Ceditest(R) and FMDV 3ABC-I-ELISA kit was 100%.However,the sensitivity of the UBI(R) kit was only 81.8%.With sera from naive or vaccinated non-infected animals,the specificity of all tests exceeded 90%.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

OBJECTIVETo explore the molecular biological mechanism of acupuncture and moxibustion (A&M) in reducing chemotherapy-related toxicity, relieving myelosuppression and increasing peripheral white blood cells (WBC).

METHODSTwo hundred and twenty-four male Kunming mice of clean grade were randomized equally into 4 groups, the blank control group (A), the model group (B), the acupuncture group (C), and the moxibustion group (D). Except those in Group A, mice were duplicated into myelosuppression model with cyclophosphamide (CTX) using the accepted method. After being modeled, mice in Group C and D were treated with acupuncture and moxibustion respectively, once a day for 7 successive days, while those in Group A and B were dealt with the same actions (seizing and fixing) every day but no therapy was given. From day 2 to day 7 of the treatment, 8 mice were taken from every group per day and killed in batches. Their peripheral WBC was counted and bone marrow for detecting Cyclin D1 expression and percentages of bone marrow cells in different cycle stages using immunohistochemistry and flow cytometer respectively.

RESULTSWBC count restored to exceed the baseline in group C and D at day 5 of the treatment, being one day earlier than that in group B. Cyclin D1 expression in the bone marrow raised in Group C and D, and reached the peak at day 4, showing significant difference as compared with that in Group B (P < 0.01). The phase G1 marrow cell percentages in Group C and D was lower than that in Group B at all days of detection, showing statistical significance at day 2-4 (P < 0.05 or P < 0.01); while the percentages of phase S and G2-M cells in the two treated groups was higher than that in group B all the times.

CONCLUSIONSWhile CTX damaged marrow cells, it intervened the cell cycle regularity and reduced the DNA content to cause myelosuppression and leucocytopenia. A&M therapy could improve the Cyclin D1 expression, speed up the cell transition from phase G1 to phase S and increase the synthesis of DNA. At the mean time, taking advantage of the block at phase G2, it can repair the injured DNA, speed up cell mitosis for turning into multiplication, improve the anti-injury and repairing ability of cells to protect bone marrow cells, and relieve the myelosuppression.

Acupuncture Therapy ; Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cyclin D1 ; metabolism ; Cyclophosphamide ; adverse effects ; Immunosuppressive Agents ; adverse effects ; Male ; Mice ; Mice, Inbred Strains ; Moxibustion

Objective To study the relationship between peripheral blood hemoglobin (HB) and blood pres- sure.Methods We performed a cross-sectional analysis in 1153 subjects aged 29-83 years.Waist circumfer- ence,HB,blood pressure,high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL- C),triglycerides (TG),total cholesterol (TC) were determined.Results ①With the increasing of blood pres- sure,HB had a clearly increasing trend (HB,normotensive:137.5?14.7 vs prehypertension:143.4?14.4 vs hy- pertension:144.3?13.8 g/L,P

OBJECTIVE: To select specific DNA aptamer for determining ketamine by FluMag-SELEX. METHODS: Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity. RESULTS: Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine. CONCLUSION FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Aptamers, Nucleotide/metabolism* ; DNA ; DNA, Single-Stranded/genetics* ; In Vitro Techniques ; Ketamine/metabolism* ; Oligonucleotides ; SELEX Aptamer Technique/methods*