Objective To preliminarily explore the early process of hepatitis B virus (HBV) infection in HepG2 cells induced by dimethyl sulfoxide(DMSO) ,and provide cytological bases for mechanism study of HBV infection in vitro .Methods HepG cells were divided into the DMSO inducing group and control group ,and were cultured 4 days by DMEM containing 1 .5% DMSO and normal DMEM respectively ;changes of cellular morphology were observed .In addition ,selected ECV304 cells as the negative con‐trol group and treated with DMSO .Cells in the three groups were incubated 2 hours with HBV positive serum after culturing 24 hours ,then trypsin digestive solution ,HepG2 cells and ECV304 cells were collected and polymerase chain reaction (PCR) was used for the determination of HBV DNA .Simultaneously ,the blank control group was set ,and the position of HBsAg on HepG2 cells were detected by using indirect immunofluorescence .Results HepG2 cell volume in the DMSO inducing group was obviously in‐creased .HBV DNA was found in HepG2 cells both in DMSO inducing group and control group ,and DMSO inducing group ex‐pressed much stronger .The results of IIF shown that green fluorescent signals of cell membrane and cytoplasm of HepG 2 cells in the DMSO inducing group were increased obviously ,while the results of HBV DNA and IIF both were negative in the negative con‐trol group .Conclusion DMSO could facilitate adsorption of HBsAg in some extent ,which are benefit for completing the process of early infection .

BACKGROUND:It has a variety of treatments for equinovarus in spastic cerebral palsy, including physical therapy, orthotic therapy, systemic anti-spasticity drug therapy, local intramuscular injection of botulinum toxin type A and a variety of surgical operations. These treatments aim at different severity of deformities and different age in children patients, and each has its own merits. OBJECTIVE:To investigate the therapeutic effect of Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition to treat equinovarus in spastic cerebral palsy. METHODS: Twenty-two children with spastic cerebral palsy combined with equinovarus were colected from 2012 to 2014, and then these children were subject to Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition. Postoperative plaster external fixation was done for 6 weeks, and then rehabilitation training was given. RESULTS AND CONCLUSION:Al the 22 patients were folowed for an average of 1-2 years, and then were evaluated according to the degree of patient’s satisfaction and the degree of foot deformity. Satisfactory results were obtained from al the patients. Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition to treat spastic cerebral palsy combined with equinovarus is a good method, and it is characterized as good appearance of the ankle and favorable foot function.

Objective To investigate the role of microRNA-490-5p (miR-490-5p) in the regulation of visceral sensitivity in rats with intestinal dysfunction.Methods The lentivirus vector system was used to construct the rno-miRNA-490-5p lentivirus expression vector.The rats were divided into normal group,diarrhea-predominant irritable bowel syndrome (IBS-D) group,lentivirus empty vector group and the siRNA silent group and the latter three groups were model groups.The efficiency of siRNA was measured by real-time polymerase chain reaction (PCR).The rats were gavaged with 10％ India ink,and then the time of first black stool,water content of feces and threshold of expansion capacity caused abdominal elevation or back arching were calculated.The visceral sensitivity of rats after miRNA-490-5p silenced was evaluated with abdominal withdrawal reflex (AWR) score by stimulating with different intensities of colonic dilatation.The abdominal electrical activity of rats stimulated by colonic distension was measured by BL-420F biological and functional experimental system.The change of the tension of rats isolated colon intestinal stimulated with acetylcholine chloride was also detected by BL-420F biological and functional experimental system.T test was used to compare the differences between the model groups and the normal group.One way analysis of variance was performed for multi-group comparison after miRNA-490-5p interfered.For comparison between two groups among multiple groups,least significant difference (LSD) method was used when the variance was equal,and Games-Howell method was used when the variance was unequal.Results The gastrointestinal propulsion time and the threshold of expansion capacity caused abdominal elevation or back arching of model groups were both lower than those of the normal group ((8.54±4.07) hvs (12.33±2.23) h,(0.56±0.08) mL vs (0.84±0.09) mL),and the differences were statistically significant (t =2.62 and 6.37,both P ＜ 0.05).After distension with 0.8 mL and 1.2 mL sodium chloride solution,the AWR scores of model groups were significantly higher than those of the normal group (3.20±0.56 vs 1.20±0.45,3.73±0.46 vs 2.60±0.55),and the differences were statistically significant (t=7.20 and 4.58,both P＜0.01).There was no significant difference in AWR score between the model groups and the normal group when distended with 1.6 mL sodium chloride solution (3.93 ±0.26 vs 3.80 ± 0.45) (P＞0.05).After miRNA-490-5p silenced,gastrointestinal propulsion time of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group was (11.12±1.01) h,(6.23±3.17) h,(6.09 ± 2.26) h and (12.36±1.97) h,and the differences among four groups were statistically significant (F=10.55,P＜0.01).The abdominal electrical activity of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group distension stimulated with 0.8 mL and 1.2 mL sodium chloride solution was (64.91 ± 10.50),(101.79 ±11.73),(80.49±1.27),(66.92±3.24) μV,and (105.09±52.40),(131.71± 16.74),(111.00±6.41) and (95.49± 4.2) μV,and the differences among four groups were statistically significant (F=16.82 and 9.14,both P＜0.05).There was no significant difference in abdominal electrical activity amplitude between silenced group and normal group ((66.92±3.24) μV vs (64.91±10.49) μV and (95.49±4.22) μV vs (105.09±2.40) μV) (all P＞ 0.05).After distension with 1.6 mL sodium chloride solution,the abdominal electrical activity amplitudeof silenced group was lower than the other groups,and the differences were statistically significant (F=11.09,P＜0.01).After adding 1∶1 000 acetylcholine chloride added,the tension of colon of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group increased by 0.71 ± 0.21,0.81±0.06,0.88±0.21 and 0.43±0.07,however there was no significant difference among the four groups (F=2.57,P =0.100).Conclusions Visceral hypersensitivity existed in rats with intestinal dysfunction.miRNA-490-5p may be involved in the regulation of visceral sensitivity.

In this study, the human umbilical vein endothelial cell model was used to study the regulating effect of lipophilic components in Salvia miltiorrhiza on angiogenesis, and explore its possible mechanism. The cell model was established to determine the effect of lipophilic components in S. miltiorrhiza on the proliferative activity and migration capacity of endothelial cells. Then the realtime fluorescence quantification PCR technology was applied to detect the changes in the gene expressions of angiogenesis-related cytokines VEGF-A, VEGF-C and MMP-9. The results showed that 5 mg x L(-1) lipophilic components in S. miltiorrhiza could inhibit the proliferation and migration of endothelial cells, and reduce the expression of VEGF-A and MMP-9 genes. It indicated that lipophilic components in S. miltiorrhiza may inhibit the proliferation and migration of endothelial cells by inhibiting the expression of VEGF-A and MMP-9 genes, so as to show the inhibitory effect on angiogenesis.
Angiogenesis Inhibitors ; chemistry ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

0.05) in the presence of ApoE ?4 allele.In all four ApoE ?4/4 homozygote patients,one female patient presented a transient delirium status three days before surgery,another male patient presented serious fluctuated delirium symptoms from the second to 17th days after operation.Conclusion The presence of ApoE ?4 allele seems not a predictator of postoperative delirium.ApoE ?4/4 homozygote patients may be more indulgent to delirium than others.

Objective To investigate whether apolipoprotein E (ApoE) genotypes is associated with postoperative delirium in aged noncardiac surgical patients. Methods Two hundreds and twelve inpatients over 65y, undergoing selective noncardiac surgeries were enrolled in the study. The patients were frequently interviewed and evaluated prospectively for delirium with the Confusion Assessment Method (CAM) during the first three postoperative days. APOE genotype was determined using multiplex amplification refractory mutation system pelymerase chain reaction (multi-ARMS PCR) technique. Results Delirium occurred in 45 patients during the first three postoperative days. Of the 212 patients, 18 (8.5%) possessed one or two ApoE 84 allele. There was no significant difference between delirious patients and non-delirious patients(6.7% : 9.0%, P >0.05) in the presence of ApoE ε4 allele. In all four ApoE ε4/4 homozygote patients, one female patient presented a transient delirium status three days be-fore surgery, another male patient presented serious fluctuated delirium symptoms from the second to 17th days after operation. Conclusion The presence of ApoE ε4 allele seems not a predictator of postoperative delirium. ApoE ε4/4 homozygote patients may be more indulgent to delirium than others.

Objective: To compare the efficacy and side effects of patient-controlled intravenous morphine with epidural single bolus morphine in postoperative pain relief. Method: Sixty patients undergoing gynecological procedures under epidural anesthesia were randomly assigned to epidural morphine(EPI)group or patient-controlled intravenous analgesia (PCIA) group. In the EPI group,2 mg of morphine was injected into epidural space at the end of operation. In PCIA group, 1 mg of morphine as a demand dose would be injected intravenously by the patient through a patientcontrolled analgesic delivery system until the pain relieved. The patients were followed up at 4, 8, 12, 24 h after operation,and the degree of pain,sedation, nausea and vomiting were assessed. Result: The total dosage of morphine was higher in the PCIA group(19.08?5.0 mg)than that in the EPI group(2mg,P

Abstract: Objective　To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods　LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results　In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion　Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.

OBJECTIVETo observe the clinical efficacy of modified Huanglian Wendan Decoction (HWD) in treating senile mild cognitive impairment (MCI) of turbid-phlegm blocking orifice syndrome.

METHODSWith a block randomized, double-blinded and controlled design adopted, the 64 patients of MCI selected from December 2007 to February 2009 were randomly and equally assigned to two groups. The treatment group was treated with HWD in dose of 200 mL, twice a day; the control group was given Aniracetam 0.2 g (for patients over 70-years-old, 0.1 g) three times a day. And the illusive medicine in dosage-form of capsule/decoction simulated to that used in the opposite group was applied. The medication and observation lasted for three months. Chinese medicine syndrome, cognition capacity (by MMSE), laboratory indexes [acetylcholine (Ach), superoxide dismutase (SOD), malondialdehyde (MDA)] and safety related indexes in patients were observed.

RESULTSAfter treatment, MMSE score increased in both groups, but the increment in the treatment group was significantly higher than that in the control group (P<0.01); Chinese medicine syndrome estimated by scoring showed that after treatment, all scores of syndromes, excepting the expectoration, were improved in the treatment group with the post-treatment scores significantly lower than those in the control group respectively (P<0.05 or P<0.01); while in the control group, lowering of scores only showed in some symptoms such as poor memory, heavy head or dizziness, and heavy sensation in limbs and body. Serum levels of Ach and SOD decreased and MDA increased in both groups after treatment, but the change of Ach was more significant in the treatment group (P<0.01). No obvious adverse reactions were found during the treatment.

CONCLUSIONFor treatment of MCI, HWD shows effects in improving patients' symptoms, cognition capacity and elevating serum Ach content better than that of Aniracetam; and with effects for raising SOD activity and reducing MDA level similar to those of Aniracetam.

Aged ; Aged, 80 and over ; Cognitive Dysfunction ; diagnosis ; drug therapy ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Pyrrolidinones ; therapeutic use

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis