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.

It is particularly important to confirm that whether subjects were repeated to participate in the clinical trial, the time interval from the last dose in the recent clinical trial, whether the physical conditions of subjects accord with the clinical trial protocol in phase I clinical trials and so on.This paper will introduce two types of the subject database, one was established by our hospital, another one is subject networking database, and will introduce the operation mode, the present situation of subjects screening under the database management. To explore the existing problems, optimize the use of subject database and give long-term prospects of subject networking database.

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

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 dose-response relationship of the treatment temperatures and heating time on human cervical carcinoma hela cells,aiming at providing experimental evidences for clinical hy-perthermia. Methods Hela cells were heated at 37 ～ 70 ℃ in temperature-controlled water baths, the tempera-ture was divided into nine groups,each time was divided into eight subgroups (1 ～ 30 min). The morphology changes of cells after hyperthermia were detected by inverted microscope. Proliferation rates were measured by MTT colorimetric assay. The apoptesis rates were determined by flow eytometric analyse. The levels of prolifera-ring cell nuclear antigen (PCNA) were measured with immunohistochemistry. Results lnereaseing the heating time at the same temperature, or increaseing heating temperature at the same time, the cell proliferation, survival rates and PCNA expression decreased. There was no significant morphological change about cells ,but have small amount of apoptosis and a direct role of the suppression and destruction at 41 ℃ and 43 ℃ group. A large num-ber of cells shrinked to round and a major role for apoptosis at 46℃ group. Cell necrosis was major role at 50 ℃and 55 ℃ group. More than 55 ℃ for necrotic cells. Conclusion With the increase of heating temperature and heating time, its treatment of Hela cells gradually enhance. So combining dose-effect relationship of hyperthermia temperature and time can reach the best therapeutic effects.

Objective To determine the diagnostic value of shoulder MR arthrography in revealing the injuries of anterior glenoid labrum,and to compare MR arthrography with arm in neutral position with MR arthrography with arm in abduction and external rotation(ABER)position.Methods MR arthrography of the shoulder,including additional oblique axial sequences with the patient in the ABER position,was performed in 44 patients.The injuries of anterior glenoid labrum of these patients were retrospectively evaluated.The result was compared with that of arthroscopy.Results For displaying the abnormalities of anterior glenoid labrum,the sensitivity,specificity,and accuracy of axial MR images with arm in neutral position were 79.3%(23/29),100%(15/15),and 86.4%(38/44),respectively.While the results of ABER position oblique axial images were 93.1%(27/29),100%(15/15),and 95.5%(42/44), respectively.The difference of sensitivity between axial and ABER-position scans was statistically significant (P

Objective To compare the detection performance of the modified serum test (TRUST) method and the traditional method in syphilis screening.Methods A series of TRUST high titer syphilis serum was diluted,and the positive rate of each method was calculated by using the improved method and the traditional method.Comparison of two detection methods of C50,C5 ～ C95 interval,as well as the accuracy of the density curve,and the consistency of the two methods were compared,and diagnostic performance were compared.Results The improved method of C50 was less than the traditional method of C50,and the improved method of C5 ～ C95 range was narrower than the traditional method,compared with the traditional method.The improved method of the non precision density curve was steeper than the traditional method,and the two confidence interval of the consistency degree of the 95％ methods was 73.4％ to 95.8％.The diagnostic sensitivity (SEN),diagnostic specificity (SPE),positive predictive value (PPV),negative predictive value (NPV) and diagnostic efficiency(DF) of the improved method were 64.29 ％,99.1％,85.71％,97.05 ％ and 96.39 ％,respectively.The SEN,SPE,PPV,NPV and DF of the traditional methods were 3.75 ％,98.49 ％,75 ％,96.18 ％ and 95 ％,respectively.The improved method was superior to the traditional methods in the two aspects of SEN and PPV (x2 =8.25,10.03,all P＜0.05),with the statistically significant difference.The improved method was slightly higher than the traditional method in SPE,NPV and DF (x2 =2.39,3.45,4.03,all P＞0.05),with the no statistically significant difference.Conclusion The precision,diagnostic sensitivity and diagnostic specificity of the improved method was higher than that of the traditional method,and it can be applied to the detection of large batch samples with the aid of the full automatic enzyme immunoassay instrument.The improved method can be used to replace the traditional method for syphilis screening.

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.