BACKGROUND: Previous studies demonstrated that eucommia bark can promote bone marrow stern cells (BMSCs) differentiated into osteoblasts, but relative mechanism is poorly understood. OBJECTIVE: To investigate the effects of eucommia bark water/methanol extracts on expressions of osteopontin (OPN) and osteoprotegedn (OPG) in rat BMSCs. METHODS: Totally 2 g eucommia bark powder were added into water or methanol to 16 mL and oscillated for 1 hour at room temperature. After soaked overnight, both extracts were centrifuged at 15 000 r/min for 10 minutes. Water extract was obtained from supernatant in water soaked powder. In methanol soaked powder, methanol extracts was obtained by concentrated supernatant in vacuo and resolved using 16 mL water. Water and methanol extracts were then filtered by 0.22 μm membrane, and conserved at -20℃. Six SD rats, aged 2 months, were selected, and the 3~(rd)passage of BMSCs were induced by water or methanol extracts with dilution of 1 × 10~(-2), 1 × 10~(-3), 1 × 10~(-4) and 1 × 10~(-5), respectively. PBS was added in the negative control group. All cells were cultured for 6 days. Expressions of OPN and OPG was measured by immunocytochemistry at 6 days with induction. The expression of OPN and OPG induced by water and methanol at 1 ×10~(-3) and 1×10~(-4) dilution was detected by RT-PCR. RESULTS AND CONCLUSION: Immunocytochemistrical results indicated that both water and methanol extracts of eucommia bark simulated OPN and OPG expression, in particular with dilution of 1×10~(-4). The methanol extracts had a stronger effect than water extract, but the expression of OPG did not change obviously. RT-PCR demonstrated that at the 3rd day of inducement, the level of OPN expression induced by water extract was higher than that of methanol extract, and no OPG expression was detected. Osteogenic differentiation of rat MSCs induced by eucommia bark water/methanol extracts relates to stimulating expression of OPN, which has no correlation to OPG. OPN expression induced by water extract is early than that of methanol extract.

Objective To explore the changes of deceleration capacity of rate (DC) and analyze its correlation with heart rate variability (HRV) and other factors in patients with coronary heart disease. Methods One hundred and twenty-nine patients with coronary heart disease (coronary heart disease group) and 109 healthy people (control group) were enrolled in this study. DC and HRV parameters were measured by using digitized 24 h Holter. The correlation between DC and HRV parameters, other factors were analyzed. Results The levels of DC, SDNN, SDANN, SDNNi, PNN50, TP, LF, HF, AC in coronary heart disease group were significantly lower than those in control group:(5.64±1.67) ms vs. (6.71±1.47) ms, (106.60±20.53) ms vs. (138.82±31.22) ms, (96.94±20.06) ms vs. (127.47±31.87) ms,(28.53±14.75) ms vs. (52.24±14.65) ms, 87.72%vs. 103.86%,(1 967.10±966.16) ms2/Hz vs. (2 846.70±1 443.41) ms2/Hz,(326.43±195.35) ms2/Hz vs.(457.64±254.30) ms2/Hz, 85.88 vs. 106.39, (-6.18±2.15) ms vs. (-7.00±2.51) ms, P<0.05 or<0.01. DC was correlated with SDNNi, PNN50,TP,LF, HF, AC both in total population or in coronary heart disease group and control group by using multiple linear correlation analysis ( r=0.586, 0.356, 0.531, 0.563, 0.435,-0.433, P<0.01). After removing confounders, DC was correlated with age, SDNNi, rMSSD, PNN50 and AC (P<0.01). Conclusions DC decreases in patients with coronary heart disease and is strong correlativity with HRV parameters. DC could be used for quantitative detection of autonomic nervous function.

OBJECTIVE: Mitochondrial deoxyribonucleic acid (mtDNA) 8344 A>G (m.8344A>G) mutation is the common mutation associated with mitochondrial myoclonus epilepsy with ragged-red fibers (MERRF) syndrome. Herein we report a rare case with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes/MERRF/Leigh (MELAS/MERRF/Leigh) overlap syndrome caused by m.8344A>G mutation. METHODS: The clinical and imaging data of the patient were collected and an open muscle biopsy was carried out. We further employed molecular genetic analyses to detect mtDNA mutation in the proband and his mother. And then a clinical and neuroimaging follow-up was performed. RESULTS: This patient was a 25-year-old male, who developed exercise intolerance since the age of 6. At age 10, he suffered from acute episodes of hemianopia, and cranial magnetic resonance imaging (MRI) showed occipital stroke-like lesions and cranial magnetic resonance spectroscopy (MRS) revealed a lactate peak corresponding to the lesion. After that the patient presented slowly progressive psychomotor decline. He had myoclonic seizures and cerebellar ataxia since the age of 12. At age 21, he was admitted to our hospital because of confusion and cranial MRI revealed symmetrical lesions in bilateral posterior putamen, thalami and midbrain. Then repeated MRI showed progression of original lesions and new frontal multiple stroke-like lesions. Symptomatic and rehabilitation treatment relieved his condition. Follow-up cranial MRI at age 24 showed the lesions in basal ganglia and thalami diminished, and the midbrain lesions even completely vanished. Muscle pathology indicated the presence of numerous scattered ragged-red fibers (RRF), suggestive of a mitochondrial disorder. Polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) detected the m.8344A>G mutation of the MT-TK gene encoding mitochondrial transfer RNA for lysine in the patient's blood. Next generation sequencing (NGS) of the whole mitochondrial genome identified that the proportion of m.8344A>G was 90%, and no other mtDNA mutation was detected. Sanger sequencing further identified this mutation both in the proband and his mother's blood, although the mutation load was much lower in his mother's blood with approximately 10% heteroplasmy. CONCLUSION The present study is the first to describe a patient with m.8344A>G mutation in association with the MELAS/MERRF/Leigh overlap syndrome, which expands the phenotypic spectrum of the m.8344A>G mutation.
Acidosis, Lactic ; Adult ; Child ; DNA, Mitochondrial/genetics* ; Humans ; Male ; Mitochondrial Encephalomyopathies ; Mutation ; Stroke ; Young Adult

Objective To evaluate the efficacy and toxicity of postoperative chemoradiation for D2 dissection gastric cancer,and to compare the difference of toxicity between confromal and traditional radiotherapy.Methods Sixty four patients with T3-4,N + or R1 were enrolled.Radioation was given to a total dose of 46Gy delivered in 23fractions by use of 3D-CRT or 4 fields traditional radiotherapy.Chemothrepy was administered with paclitaxel 135 mg/m2 day 1 and 29,cisplantin 20 mg/m2 day 1 ～3 and day 29 ～31 during radiotherapy.Results The median follow-up time was 40 months.The 3-year overall and relapse-free survival rates were 78.2% and 70.9%,respectively.Eighteen patients had tumor relapse.Fifty-three patients completed chemoradiotheray.Toxicities on grade 3 or above included gastrointestinal toxicity (28.1% ),eutropenia (21.8 % ) and alopecia ( 18.7% ).One patient died of hemorrhage of upper digestion tract.Conclusion Adjuvant radiotherapy with paclitaxol and cisplatin yielded satisfactory overall survival and disease-free survival in gastric cancer patients.The toxicity was manageable.Conformal radiotherapy seems to decrease the gastrointestinal toxicities compared to that occurred in the traditional radiotherapy.

OBJECTIVETo analyse dental crown fracture occurrence situation in the long-term mastication process, and discuss the correlation factor of crown fracture occurrence.

METHODSFrom December, 2005 to December, 2007, 255 crown fracture teeth were chosen as the object of study. The sex, age, teeth position, fracture position and occlusal state of patients were analyzed.

RESULTSIn 255 teeth, 1) 76 teeth (29.8%) were maxillary first molars, 45 teeth (17.6%) were mandibular first molars, 41 teeth (16.1%) were maxillary second molars, 37 teeth (14.5%) were mandibular second molars, 32 teeth (12.5%) were maxillary second premolars, 15 teeth (5.9%) were maxillary first premolars, 9 teeth (3.5%) were mandibular second premolars. 2) The fracture position of 158 teeth (62.0%) overlapped with pit and fissure. The fracture position of 97 teeth (38.0%) occurred in dental inclined surface. 3) The occlusal state of 85 patients (33.3%) was normal. The occlusal surface shape of 55 patients (21.6%) was unnormal. The occlusal shape of 115 patients (45.1%) was unnormal.

CONCLUSIONThe teeth position and occlusal state has certain relations with occurrence of dental crown fracture. Occlusal force was the basic factor for dental crown fracture, but it was not the only one.

To construct a plasmid expressing MDR1 short hairpin RNA (shRNA) mediated by pEGFP-C1/U6 vector, two coding sequences of 19 nucleotides were selected from MDR. Two pairs of oligonucleotides were designed for these two fragments. After annealing the formed double-stranded DNAs were ligated with plasmid pEGFP-C1/U6 (pEGFP-C1 vector with U6 promoter). The plasmids producing MDR1 shRNA were constructed from the inverted motif containing 9 spacers and four Ts. The results showed that the constructed plasmids were named pEGFP-C1/U6/A and pEGFP-C1/U6/B, and the constructs were identified by restriction and sequence analysis, no any base mutation was observed. It is concluded that plasmids of pEGFP-C1/U6/A and pEGFP-C1/U6/B expressing MDR1 shRNA were successfully constructed, providing a highly effective method for reversing the multidrug resistance in clinic.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; DNA-Binding Proteins ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; genetics

Objective To retrospectively explore the incidence and causes of apnea after operation for retinopathy of prematurity(ROP). Methods The clinical data of 17 premature infants with operation for ROP(ROP group)and the other 23 premature infants without ROP(control group)were collected.The occurrence of apnea(time of onset and remission and original diseases) was recorded after operation in ROP group and at adjusted 37 weeks of gestational age in control group.Follow-up was conducted for 4 weeks,and the prevalences of apnea were compared between these two groups.Results There was significant difference in prevalenees of apnea between ROP group and control group ( 52.9%vs 21.7%,P<0.05).In ROP group,time of occurrence of apnea was (38±40)h after operation,and time of remission was (126±145)h after onset.Among the infants with apnea,there were 3 cases of pneumonia(33.3%),1 case of hypoglycemia(11.1%)and 5 cases with unexplained causes(55.5%). Conclusion Infants after operation for ROP are more prone to apnea,and pediatricians and ophthalmologists are required to collaborate in the perioperative care.

Objective To investigate the relationship between apolipoprotein E(Apo E) gene poly-morphism and mild cognitive impairment (MCI) in patients with type 2 diabetes mellitus (T2DM), and e-valuate the correlative risk factors. Method 40 cases of type 2 diabetes with MCI and 80 cases of type 2 diabetes without MCI were enrolled in this study. The polymorphism of the Apo E gene was detected by PCR-restriction fragment length polymorphism(PCR-RFLP). According to the clinical data such as course of disease, plasma glucose, plasma fat and body mass index (BMI), the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression. Results The frequency of Apo E ε_4 allele in the group of type 2 diabetes with MCI was higher than that without MCI ( 25.0% vs 10. 0% ), and the difference had statistical significance( P < 0. 01 ). The indexes of the statistical significant difference be-twcen the two groups were age, course of disease, postprandial blood glucose ( P2BG), HBA1C, BMI,family history of T2DM, hypertension, diabetic retinopathy, diabetic peripheral neuropathy, Apo E gene. The independent risk factors included diabetic retinopathy ( OR = 3. 452, P < 0. 05 ), diabetic peripheral neuropathy( OR = 3. 252, P <0. 05), Ape E gene( OR = 2. 441, P < 0.01 ), HBA1C ( OR = 1. 372, P <0.05), P2BG(OR = 1. 194, P <0.05), age(OR = 1. 194, P <0.01) and course of disease(OR =1. 142, P <0. 05). Conclusion Apo E ε_4 allele has significant relationship with T2DM and MCI. The age, course of disease, control of plasma glucose, and microvascular complication of diabetes have relation-ship with the cognitive function.

To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.
Base Sequence ; Gene Amplification ; Genome, Viral ; Human bocavirus ; chemistry ; classification ; genetics ; Humans ; Molecular Sequence Data ; Nucleic Acid Conformation ; Parvoviridae Infections ; virology ; Phylogeny ; RNA, Viral ; chemistry ; genetics ; Terminal Repeat Sequences

Objective:To investigate the relationship between tumor necrosis factor-α (TNF-α)-308A/G gene polymorphism and mild cognitive impairment(MCI)in patients with type 2 diabetes mellitus(T2DM),and their correlative risk factors thereof.Methods:Forty cases of T2DM with MCI and 80 cases of T2DM without MCI were selected for this study.The polymorphism of the TNF-α-308A/G was detected by PCR-restriction fragment length polymorphism (PCR-RFLP).According to the clinical data,such as course of disease,plasma glucose,plasma fat and body mass index(BMI),the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression.Results:The frequency of TNF-α2 allele was significantly higher in the group of T2DM with MCI than that without MCI (P＜0.01).The indexes of the statistical significant difference between the two groups were the age,course of disease,postprandial blood glucose(P2BG),glycosylated hemoglobin,body mass index,family history of T2DM,hypertension,diabetic retinopathy,diabetic peripheral neuropathy and TNF-α.The independent risk factors included TNF-α,diabetic peripheral neuropathy,diabetic retinopathy,age and P2BG.Conclusion:There is a significant relationship between TNF-α2 allele and T2DM with MCI.There is a significant relationship between the age,control of plasma glucose and microvaseular complication of T2DM with the cognitive funotion.