AIM To establish an HPLC method for the simultaneous content determination of swertiamarin,gentiopicrin,swertianolin,taxifolin,silychristin,silydianin and silybin in Dangfei Liganning Capsules (Silybi Fructus and Swertiae Herba).METHODS The analysis of 75％ methanol extract of this drug was performed on a 30 ℃ thermostatic Kromasil C18 column (4.6 mm × 250 mm,5.0 μm),with the mobile phase comprising of methanol-0.1％ formic acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelengths were set at 254 nm and 288 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.78％-100.45％ with the RSDs of less than 2.0％.CONCLUSION This accurate and reproducible method can be used for the rapid quality control of Dangfei Liganning Capsules.

OBJECTIVETo investigate the role of autophagy in acute lung injury (ALI) caused by multiple trauma in rats via pretreat with 3-methyladenine (3-MA).

METHODSForty-five Sprague-Dawley male rats, with age of 4 months and body weight of 250-300 g,were randomly divided into three groups. In the sham group, the rats received sphenotresia only;in the control group, the rats were made model of femur shaft fracture combined with brain injury, and treated with physiological saline by abdominal cavity at 1 hour before making model; in the 3-MA group, the rats were made model of femur shaft fracture combined with brain injury,and treated with 3-MA of 10 mg/kg by abdominal cavity at 1 hour before making model. Histologic changes and the concentration of related inflammatory factors in the damaged lung tissue were examined at 48 h after opteration, at the same time, the effect of 3-MA on the expression of LC-3 II and Beclin-1 was examined through reverse transcriptase polymerase chain reaction technique (RT-PCR).

RESULTSCompared with sham group, LC-3 II and Beclin-1 level in control group at 48 h after operation were obviously increased (P < 0.01). Compared with control group, LC-3 II and Beclin-1 level in 3-MA group at 48 h after operation were obviously decreased (P < 0.01). Compared with sham group, the level of proinflammatory cytokines (TNF-α and IL-6) in control group obviously enhanced (P < 0.01). Compared with control group, above items in 3-MA group was obviously lower (P < 0.01). Compared with control group,the histopathological damage of lung in 3-MA group obviously reduced (P < 0.01).

CONCLUSIONAutophagy can aggravate the acute lung injury caused by fracture of shaft of femur combined with brain injuries,but 3-MA can reduce tissue damage by inhibiting the autophagy and inflammatory response.

Acute Lung Injury ; prevention & control ; Adenine ; analogs & derivatives ; therapeutic use ; Animals ; Apoptosis Regulatory Proteins ; analysis ; Beclin-1 ; Interleukin-6 ; analysis ; Lung ; chemistry ; immunology ; pathology ; Male ; Multiple Trauma ; complications ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

Objective To isolate cancer stem cells from human colon cancer cell line CW-2,and observe the biological characteristics.Methods The percentage of CD44＋EpCAM＋ cancer stem cells which were isolated by multi-combination method was evaluated by flow-cytometry.The biological characteristics of the stem cells were analyzed by cell cycle analysis,in vitro invasion assay and in vivo tumorigenicity assay.Results The percentage of CD44＋EpCAM＋ cancer stem cells was 89.57%,and their capabilities of generation,invasion,tumorigenicity were higher than non-cancer stem cells（P 0.05）.Conclusion CD44＋EpCAM＋ cancer stem cells showed low generation,high invasiveness,high tumorigenesis,and they could be isolated by multi-combination.

Objective The study was designed to investigate the serum levels of HE4 in patients with lung cancer,and explore its prognostic value.Methods Blood samples of 106 healthy adults and 191 patients with lung cancer before treatment were measured by means of ELISA for HE4 levels in different stages and pathological types,for analyzing prognostic effect of HE4.Results The serum levels of HE4 in patients with lung cancer (median level 91.63 pmol/L) were significantly higher than control group (median level 56.42 pmol/L) (U =3 081.00,P ＜ 0.05).AUC of serum HE4 was 0.85,with a cut-off value of 82.70 pmol/L (specificity =95.31％,sensitivity =62.32％).HE4 expression was not statistically related to clinical stage and pathological types of lung cancer.The median overall survival (mOS) was 36.87 months in the HE4-low group and 30.43 months in the HE4-high group (P ＜0.05),respectively.HE4 level was an independent prognostic factor for overall survival (HR =2.15,95％ CI 1.49-3.12,P ＜ 0.05).Conclusions The prognosis of patients with high HE4 expression is poor.Therefore,it might be necessary for patients with high level of HE4 to receive more aggressive adjuvant therapy.

Objective To explore the effects of upper limb robot-assisted therapy on motor function and activities of daily living in con-valescent stroke patients. Methods From June to September, 2016, 12 chronic stroke patients at their first-ever stroke were enrolled and ran-domized into experimental group (n=6) and control group (n=6). Both groups received routine rehabilitation. Additional robot-assisted thera-py was provided to the experimental group, and additional repetitive movement training was provided to the control group, 20 minutes a day, five days a week for four weeks. Fugl-Meyer Assessment-Upper Extremities (FMA-UE), modified Ashworth Scale (MAS) and Func-tional Independent Measure (FIM) were used to assess the motor function of the upper limbs and hands, the muscular tension of shoulder and elbow, and activities of daily living (ADL) before and after treatment. Results After treatment, the scores of FMA-UE and FIM were bet-ter in both groups (Z>2.032, P<0.05), and no significant difference was found between two groups (t<0.723, P>0.05), however, the scores were a little bit higher in the experimental group than in the control group. After treatment, for the experimental group, the MAS scores of shoulder abduction/adduction and elbow flexion/extension improved (Z>2.121, P<0.05);for the control group, the MAS scores of shoulder abduction/adduction improved (Z>2.000, P<0.05), but the MAS scores of elbow flexion/extension were not significantly different (Z<1.890,P>0.05). There was no significant difference in the MAS scores of shoulder abduction/adduction and elbow flexion/extension between two group (Z<1.734, P>0.05). The moving trail recorded by the computer, gradually became a regular pattern from the mass, saying the motor control ability became better. Conclusion Upper limb robot-assisted therapy can promote the recovery of the motor function of upper limbs and ADL in convalescent stroke patient, similar to the repetitive movement training.

Objective To evaluate the diagnostic performance of Xpert MTB/RIF for the diagnosis of tuberculous meningitis (TBM).Methods This was a prospective, single center clinical trial.A total of 116 consecutive patients with suspected meningitis who were admitted to Xijing Hospital from October 2013 to June 2015 were recruited.Mycobacterium tuberculosis ( MTB) and rifampicin ( RIF) resistance mutations in 1 ml cerebrospinal fluid ( CSF) were detected with Xpert MTB/RIF and the remaining sample was tested by Ziehl-Neelsen staining , MGIT960 liquid culture and other laboratory tests .And the enrolled patients were grouped according to the 2010 South African expert consensus .The diagnostic performance of Xpert MTB/RIF was evaluated by comparing against clinical score >5 points and MGIT960 liquid culture as reference standards respectively .The comparison was performed using a χ2 test or Fisher′s exact test for categorical variables and a nonparametric rank sum test for continuous variables .Results Among the enrolled 116 subjects, 23 subjects were diagnosed as definite-TBM by MGIT960 liquid culture, 16 subjects were classified as probable TBM , 27 subjects were classified as possible TBM , and 50 subjects were classified as non-TBM.When clinical score >5 points was used as a reference standard , the sensitivity of Xpert MTB/RIF (39.4%) was comparable with that of MGIT960 liquid culture (34.8%) (χ2 =0.292, P=0.589), and significantly better than that of Ziehl-Neelsen staining (9.9%) (χ2 =16.500, 12.771, P<0.001). No significant differences were found among the specificities of Xpert MTB /RIF, MGIT960 liquid culture and Ziehl-Neelsen staining ( 98.0%, 100.0% vs 98.0%, χ2 =1.014, P=0.602 ) .When tested against MGIT960 liquid culture as a reference standard , the sensitivity of Xpert MTB/RIF was 91.3%. Conclusions Xpert MTB/RIF is a rapid and specific method to detect MTB and RIF resistance in CSF .It exhibits a good rule in value for the diagnosis of TBM and a comparable sensitivity with MGIT 960 liquid culture, thus it can be used as the initial method for the diagnosis of tuberculous meningitis .

OBJECTIVETo discuss the protective effect of alkaloids from Piper longum (PLA) in rat dopaminergic neuron injury of 6-OHDA-induced Parkinson's disease and its possible mechanism.

METHODThe rat PD model was established by injecting 6-OHDA into the unilateral striatum with a brain solid positioner. The PD rats were divided into the PLA group (50 mg x kg(-1) x d(-1)), the madorpa group (50 mg x kg(-1) x d(-1)) and the model group, with 15 rats in each group. All of the rats were orally given drugs once a day for 6 weeks. Meanwhile, other 15 rats were randomly selected as the sham operation group, and only injected with normal saline in the unilateral striatum. The behavioral changes were observed with the apomorphine (APO)-induced rotation and rotary rod tests. The number of tyrosine hydroxylase (TH)-positive cells in rat substantia nigra and the density of TH-positive fibers in striatum were detected by tyrosine hydroxylase immunohistochemistry. The content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), catalase (CAT), malondialdehyde (MDA), nitric oxide (NO) and nitric oxide synthase (NOS) in rat substantia nigra and striatum were measured by the spectrophotometric method.

RESULTAfter being induced by APO, PD rats showed obvious rotation behaviors, with decreased time stay on rotary rod and significant reduction in the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. The activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity significantly decreased, whereas the activities of NOS and the content of MDA, NO significantly increased. PLA could significantly improve the behavioral abnormality of PD rats and increase the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. It could up-regulate the activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity, and decrease the content of NOS and the content of MDA, NO.

CONCLUSIONAlkaloids from P. longum shows the protective effect in substantia nigra cells of 6-OHDA-induced PD model rats. Its mechanism may be related with their antioxidant activity.

Administration, Oral ; Alkaloids ; administration & dosage ; pharmacology ; Animals ; Apomorphine ; pharmacology ; Catalase ; metabolism ; Dopamine Agonists ; pharmacology ; Dopaminergic Neurons ; drug effects ; metabolism ; pathology ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Motor Activity ; drug effects ; Neostriatum ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; physiopathology ; prevention & control ; Piper ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism ; Tyrosine 3-Monooxygenase ; metabolism

Objective To compare the spine intensity modulated radiation therapy (IMRT) and the conventional radiation therapy on the beagle spinal cord neurons,in order to prove the biological safety of IMRT of the spinal cord.Methods Twelve selected purebred beagles were randomly divided into 2 groups.A beagle clinical model of tumor was mimiced in the ninth and tenth thoracic vertebrae.Then the beagles were irradiated by 2 different models of intensity modulated radiotherapy and conventional radiation therapy,with the total irradiation doses of 50 and 70 Gy.The samples of spinal cord were taken out from the same position of the nine and tenth thoracic vertebrae at the third month after radiation.All the samples were observed by the electron microscope,and the Fas and HSP70 expression in spinal cord neurons were evaluated by immunohistochemistry method.Terminal deoxynucleatidyl transferase mediated dUTP nick and labeling (TUNEL) technique was used to examine the apoptotic cells in the spinal cord.Results The neurons in the spinal cord of IMRT group were mainly reversible injury,and those in the conventional radiation therapy were mainly apoptosis.Compared with the conventional radiation therapy group [50 Gy group,(7.3±1.1)% ;70 Gy group,(11.3 ±.4)%],the apoptosis rate of the spinal cord neurons of the intensity modulated radiotherapy group[50 Gy group,(1.2 ±0.7)%;70 Gy group(2.5 ±0.8)%] was much lower[(50 Gy group,t＝0.022,P＜0.05;70 Gy group,t＝0.017,P＜0.05)].The expression levels of Fas in the IMPT group(50 Gy group,4.6 ±0.8;70 Gy group,7.4 ±1.1)were also much lowerthan those in the other group(50 Gy group,15.1 4-6.4;70 Gy group,19.3 ±7.6.50 Gy group,t＝0.231,P＜0.05;70 Gy group,t:0.457,P＜0.05),while the expression levels of HSP70 in the IMPT group(50 Gy group,9.1 ±0.8;70 Gy group,7.3±1.4)were much higher than those in the conventional radiation therapy group(50 Gy group,2.1 ±0.9;70 Gy group,1.7±0.3;50 Gy group,t＝0.153,P＜0.05;70 Gy group,t＝0.223,P＜0.05).The expression of Fas/HSP70 was positively correlated with the apoptosis rate of the spinal cord neurons(r＝0.996,t＝1.14.P＜0.05).Conclusions The late radiotherapy response of the spinal cord neurons was obviously observed on the third month.Comparison of the morphology and apoptosis rate of the spinal cord neurons after radiotherapy,the expression of Fas and HSP70 indicated that the biological safety in the 1MRT might be much better than that the conventional radiation therapy.

Objective To explore the causes of postoperative recurrence in patients of nodular goiter,the selection of method for reoperation and the postoperative complications. Methods The clinical data of 278 nodular goiter patients admitted in the first affiliated hospital of Harbin Medical University between 2001 and 2008 were analyzed retrospectively,including the methods and complications for first operation and reoperation. Results In the first operation,79 eases received simple eminectomy and 167 cases received partial lobectomy,accounted for 28.4% and 60.1%,respectively. Unilateral subtotal lobectomy plus contralateral eminectomy was performed in 23 cases and subtotal thyroideetomy was conducted in 9 cases,accounted for 8.3% and 3.2%,respectively. Postoperative complications occurred in one hundred and twenty-three cases,the incidence being 8.2% (23/278). Unilateral subtotal Iobectomy plus contralateral partial iobectomy was reperformed in 37 cases and bilateral subtotal thyroidectomy in 241 cases. Postoperative complications occurred in 12 cases,the incidence being 4.2%(12/278). No postoperative recurrence of nodular goiter was found. Conclusions Recurrence of nodular goiter is closely associated with the scope of previous surgical treatment,and correct operative manipulation may reduce the recurrent rate.