Objective To investigate the effects of Aktl gene knockout on pain behavior induced by chronic constriction injury model of sciatic nerve (CCI).Methods C57BL/6 male mice were randomly divided into Akt1 knockout group (KO group,n=12),wild type group(WT group,n=12).All mice were made model of CCI in the right sciatic nerve.Each mouse received tests of the paw withdrawal mechanical threshold (PWMT) and the paw withdrawal thermal latency(PWTL) at the times of 1d before and 1 d,3 d,5 d,7 d,10 d,14 d,17 d,21 d after surgery.Results For both KO group and WT group,the basic values of PMWT(right(0.89±0.15)g,(0.87±0.15)g; left(0.97±0.19) g,(1.05±0.14) g,P＞0.05) and PWTL(right (7.64±0.71) s,(7.56±0.68) s ;left: (7.67±0.6) s,(7.64±0.64) s,P＞0.05) showed no significantly statistical difference.Compared with WT group and the basic value,PWMT and PWTL were significantly decreased after surgery in KO group (P＜0.05).The PWMT and P WTL of the left paw in KO group and WT group had no obvious statistical difference (P＞0.05).However,the PWMT and PWTL of the right paw significantly decreased in the two groups compared with left paw.Conclusion h aggravates the neuropathic pain induced by CCI in mice when the Akt1 gene was knocked out.

BACKGROUND:Studies have demonstrated that Acorus tatarinowi and its active ingredients can promote adult neurogenesis, exerting an active role in anti-aging and neurodegenerative disease treatment.
OBJECTIVE:To explore the effects of Acorus tatarinowi extracts on the proliferation and osteogenic differentiation of umbilical cord blood stem cel s, thereby providing a new idea for promoting the osteogenic differentiation of stem cel s by Chinese medicines.
METHODS:Acorus tatarinowi extracts were obtained via solvent extraction method and flow cytometry sorting technology was used to select the stem cel s isolated from human umbilical cord. Then, the umbilical cord blood stem cel proliferation was observed by electron microscope, and the effect of Acorus tatarinowi on the proliferation of umbilical cord blood stem cel s was observed by cel counting kit-8. Meanwhile, the impact of Acorus tatarinowi on osteocalcin and bone morphogenetic protein-2 contents in the supernatant of umbilical cord blood stem cel s were detected by ELISA;alkaline phosphatas expression was detected using alkaline phosphatase staining kit.
RESULTS AND CONCLUSION:The separation purity of the stem cel s from umbilical cord mononuclear cel s was (89.66±3.47)%. After low, moderate and high concentrations of cord blood stem cel s co-cultured with Acorus tatarinowi extractions for 24, 48 and 72 hours, the stem cel proliferation rate was significantly higher compared with the control group (P<0.05), and additional y, the proliferation rate of moderate concentration group was significantly higher than that in the low and high concentration groups (P<0.05). The contents of osteocalcin and bone morphogenetic protein-2 in the stem cel supernatants were significantly higher than those in the control group after co-cultured with Acorus tatarinowi extractions for 5, 10 and 15 days, and which the highest in the high concentration group (P<0.05). The expression of alkaline phosphatase was significantly higher than that in the control group after umbilical cord blood stem cel s co-cultured with Acorus tatarinowi extractions for 10 days, and moreover, the expression of alkaline phosphatase in the moderate concentration and high concentration groups were significantly higher than that in the low concentration group (P<0.05). In conclusion, Acorus tatarinowi can promote the proliferation and osteogenic differentiation of umbilical cord blood stem cel s.

Objective To evaluate the role of spinal dopamine D2 receptors in a rat model of neuropathic pain.Methods Thirty healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,wcre randomly divided into 5 groups (n =6 each) using a random number table:control group (group C),sham operation group (group S),neuropathic pain group (group NP),normal saline group (group N) and dopamine D2 receptor agonist quinpirole group (group Q).Neuropathic pain was produced by chronic constriction injury of the sciatic nerve (CCI) in rats anesthetized with intraperitoneal 2％ pentobarbital sodium 40 mg/kg.At 7 days after CCI,normal saline 10 μl was injected intrathecally over 30 s in group N,and quinpirole 10 μg (in 10 μl of normal saline) was injected intrathecally over 30 s in group Q.At 1 day before CCI,3 and 7 days aher CCI,and 30 min and 1,2,4,8 and 16 h after administration,mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.Results There was no significant difference in MWT and TWL at each time point between group C and group S.MWT was significantly lower,and TWL was shorter at T1-8 in NP,N and Q groups than in C and S groups.Compared with group N,no significant change was found in MWT and TWL at each time point in N group,and MWT was significantly increased,and TWL was prolonged at T4-6 in group Q.Conclusion Inhibited function of spinal dopamine D2 receptors is involved in the maintenance of neuropathic pain in rats.

Objective To investigate the effects of repeated intrathecally kinesin superfamily protein 17 (KIF17) antisense oligodeoxynucleotide (ODN) on the expression of mLin10 and NR2B in spinal cord in a mouse model of bone cancer pain.Methods Fifty-six male C3H/HeJ mice,aged 4 ～ 6 weeks,weighting 20 ～ 25 g,were randomly divided into two groups:sham operation group (group S,n=20) and bone cancer pain group (group T,n=36).20μl α-minimal essence medium (α-MEM) which containing 2× 105 NCTC2472 osteosarcoma cells was injected into the intramedullary space of the right femur in group T.In group S,no cancer cell was instead.The number of spontaneous flinches (NSF) and the paw withdrawal mechanical threshold (PWMT) were measured at the day before (base) and the days 4,7,10 and 14 after inoculation.According to the corresponding time points,twenty-four mice were sacrificed for determination the expression of KIF17,mLin10 and NR2B using Western blot.Then,the mice of group T were randomly divided into three groups (n=8,T1,T2,T3,group).In group S and group T1,Saline 5 μl was injected intrathecally.KIF17 sense ODN and antisense ODN,5 μg/5μl were respectively injected in group T2 and T3 for 6 consecutive days.Pain behaviors were assessed at the days 2-6 after the first injection.And determinated the KIF17,mLin10 and NR2B expression,again.Results Compared with group S,the NSF was increased and the PWMT was decreased at the days 7,10 and 14 after inoculation in group T (P＜0.05).Compared with the base ((0.65±0.15),(1.06±0.06),(1.01±0.14)),the expression of KIF17,mLin10 and NR2B (14d:(1.13 ±0.06),(2.17 ± 0.37),(1.85 ± 0.32)) were increased at the days 7,10 and 14 after inoculation in group T(P＜0.05).During the course of the injection,compared with group T1 and T2,the NSF was decreased and the PWMT was increased significantly in the group T3(P＜0.05),the expression of KIF17,mLin10 and NR2B((0.88±0.08),(0.96±0.11),(1.03±0.08)) were reduced in group T3 (P＜0.05).Conclusion Intrathecal KIF 17 antisense ODN in the mice of bone cancer pain improves the pain behaviors,and inhibits the up-regulated of KIF17,mLin10 and NR2B during the course of the injection.

Objectives To design an objective and structured evaluation system for the clinical competence of orthopedic postgraduates in the diagnosis and treatment of distal radius fractures, and to ana-lyze its reliability and validity. Methods 28 orthopaedic postgraduates representing six levels of surgical training were tested for competence in performing surgical approach for distal radius fracture on cadaver specimens during which four measures were used to assess competency: examination of basic theory based on network item bank, objective structured operation assessment,overall assessment and operation examina-tion results. In addition, the time for completion of the surgery was also recorded. Each assessment tool was correlated with the others as well as with the resident’s level of training. Results There was a significant correlation between the seniority of candidates and the score of theoretical examination (F=6.193, P=0.000), the score of structured operation examination (F=6.374, P=0.002), the score of overall assessment (F=2.321, P=0.030), and the passing rate of final operation examination (F=36.300, P=0.000). No significant differ- ences were found between seniority and time to completion of the surgical approach exposure (F=2.282, P<0.073). Conclusions The results of the present study suggested that both theoretical examination and cadaver testing discriminate between novice and accomplished postgraduates. However, although the theo-retical test scores could predict the operational test results, but the theoretical results can not guarantee excellent operational skills.

Objective To explore the application of mixed anesthesia with epinephrine hydrochloride and lidocaine in se-nile ptosis surgery.Methods 90 senile ptosis cases(180 eyes)treated from January 2016 to June 2016 were randomly divided into experimental group and control group.Subcutaneous infiltration anesthesia with lidocaine and epinephrine hydrochloride was used in the experimental group,while the control group received lidocaine only.The heart rate,blood pressure,blood loss,visual field definition,operation time,incision state,cornea and recovery rate of the two groups were observed.Results① The systolic blood pressure of the patients in the experimental group was significantly higher than that of the control group (P<0.05).There was no significant difference between two groups in diastolic blood pressure and heart rate(P>0.05).②The experimental group achieved better results than the control group in operation time,anesthesia time,amount of bleeding, visual field clarity,eyelid congestion and wound healing(P<0.05);③ There was no significant difference(P> 0.05)be-tween two groups in incisional dressing bleeding,fluid seepage and cornea status.Conclusion The mixed anesthesia of epi-nephrine hydrochloride and lidocaine had little effect on the patients' diastolic blood pressure and heart rate.It reduced the blood loss during operation and the postoperative eyelid congestion.The clarity of intraoperative vision and recovery time were also improved with the mixed anesthesia.

Objective: To assess the status of handgrip strength of elderly population from longevity areas in China, and to analyze the correlative factors of handgrip strength of elderly people. Methods: Data from Chinese Longitudinal Healthy Longevity Survey (CLHLS) in 2012 was used, from which1 967 participants aged ≥60 years old with valid data of grip strength value from 8 Chinese longevity areas were included. Information on demographics characteristic, life style and health status was collected using questionnaires. The handgrip strength of both left and right hands were measured by grip dynamometer. The different characteristics of group of participants with different grip strength were compared and then analyzed by adopting the Cumulative odds Logistic regression model to identify main factors associated with hand grip strength. Results: The P₅₀ (P₂₅-P₅₀) of hand grip strength of elderly people from the eight longevity areas was 20 (11-28) kg; The hand grip strength of males was 26 (18-34) kg, which was higher than that of females(14 (9-20) kg) (P<0.001). Cumulative odds Logistic regression model showed that the hand grip strength of females was lower than males, whose β value (95%CI) was-1.22 (-1.43--1.00). The elderly who was at a higher age, smoking, drinking or with anemia, had a comparatively lower handgrip strength, whose β (95%CI) value were separately-0.08(-0.09~-0.07),-0.29(-0.56~-0.02),-0.54(-0.80~-0.28), and-0.41(-0.62~-0.20). And the elderly who had a higher boby mass index, drinking tea and outdoor activities, had a comparatively higher handgrip strength, whose β(95%CI) value were separately 0.28 (0.15-0.40), 0.25(0.03-0.47) and 0.51(0.30-0.71). Conclusion Age and gender were the main correlative factors, lifestyles and physical conditions might also be correlative factors of hand grip strength of the elderly from longevity areas in China.

OBJECTIVETo study genotype-phenotype correlation of a family with late infantile metachromatic leukodystrophy(MLD).

METHODSClinical data were collected and ARSA gene was tested by PCR and sequencing in a pedigree.

RESULTSThe male proband onset with walking dysfunction at 19 months, arylsulfatase A activity of leucocyte from his peripheral blood was 20.2 nmol/mg.17h, and his cranial MRI showed wildly symmetrical demyelination. Homozygosis for novel c.622delC (p.His208Metfs46X) in exon 3 of ARSA gene was identified in proband, and heterozygous for the same mutation in parents and grandma of the proband.

CONCLUSIONLate infantile metachromatic leukodystrophy is characterized by rapid and progressive regression of neuropsychiatric and motor development. There is a significant correlation between the mutation of c.622delC(p.His208Metfs*46) in the ARSA gene and the phenotype presenting as O/O patients.

Base Sequence ; Cerebroside-Sulfatase ; deficiency ; genetics ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Infant ; Leukodystrophy, Metachromatic ; diagnostic imaging ; enzymology ; genetics ; Magnetic Resonance Imaging ; Male ; Mutation ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; Radiography ; Sequence Deletion

Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.

Objective: To explore the association between biomarkers and activities of daily living (ADL) in the elderly over 65 years old from longevity areas in China. Methods: A total of 2 439 people from 8 longevity areas were included in our baseline survey in 2012. Using questionnaires, body measurements, and blood biochemical examinations, information on demographics characteristic, life style, ADL, blood pressure and biomarkers were collected. Based on these six items of ADL (bathing, dressing, indoor activities, toileting, eating, bowel and bladder control), we constructed a dichotomous indicator for ADL. A respondent was defined as ADL disabled if any difficulty in one or more of the above six activities was reported. Information were collected in the follow-up in 2014 using the same questionnaires and examinations. We excluded information on the elderly who lacked ADL or biomarkers test results or with ADL disability at baseline study. Finally 938 elderly people over 65 years old were included in this analysis. Multivariate logistic regression model was used to analyze the influence factors of ADL disability. Results: During the 2-year follow-up, 100 (10.7%) participants developed into ADL disability, with a rate at 10.7%. Multivariate logistic regression analysis indicated that each year increase in age or each 1 mmHg (1 mmHg=0.133 kPa) increase in systolic blood pressure (SBP) would cause the risk of ADL disability to increase 9% or 1%, whose OR (95%CI) were separately 1.09 (1.06-1.12), 1.01 (1.00-1.02). Han nationality or cognitive impairment increased the risk of ADL disability, whose OR (95%CI) values were separately 4.90 (1.13-21.24), 2.47 (1.44-4.25), while increased lymphocyte count (>1.60×10⁹/L), being married, or participating in recreational activities decreased the risk of ADL disability, whose OR (95%CI) values were separately 0.51 (0.31-0.82), 0.52 (0.28-0.96), 0.43 (0.23-0.80). Conclusion In the elderly elevated lymphocyte count was associated with lower risk of ADL disability. In addition, incresed age, increased SBP, unmarried, Han nationality or cognitive impairment were associated with the increasing risk of ADL disability in older people, while participating in recreational activities would reduce the risk.