OBJECTIVE:To establish the quality standard for Chaige ganmao mixture. METHODS:TLC was conducted to iden-tify the Scutellaria baicalensis and Paeonia lactiflora;HPLC was conducted to determine the content of puerarin. The column was Zorbax Eclipse XDB-C18 with mobile phase of methanol-water(25:75,V/V)at flow rate of 1.0 ml/min,detection wavelength was 250 nm,and column temperature was 25℃,and the injection volume was 5μl. RESULTS:TLC showed clear spots and good sepa-ration. The linear range of puerarin was 18-198 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were low-er than 2%,recovery was 99.04%-99.75%(RSD=0.28%,n=6). CONCLUSIONS:The standard can be used for the quality con-trol of Chaige ganmao mixture.

Objective To investigate the expression and clinical significance of UL16 binding protein 3 (ULBP3) in human esophageal squamous cell carcinoma (ESCC) and its correlation with nature killer (NK) cells.Methods The relative expression of ULBP3 in the ESCC tissues and corresponding carcinoma adjacent tissues of 40 patients was detected by realtime-poly merase chain reaction (PCR),immunohistochemical staining and Western blot methods.The percentage of NK cells in peripheral blood of same patients was examined by flow cytometry.The correlation between ULBP3 and the percentage of NK cells was analyzed with Pearson method.Results The expression of ULBP3 at mRNA level in the tumor tissues ([4.96 ±-6.11]×10-3) was significantly higher than that of corresponding carcinoma adjacent tissues ([1.64 ± 2.96]× 10-3,t =3.656,P＜ 0.01).The immunohistochemical staining results indicated that the positive rate of ULBP3 in the tumor tissues was 60％ (24/40),however that of corresponding carcinoma adjacent tissues was only 32.5％(13/40,t=3.921,P＜0.01).The Western blot results indicated that the expression of ULBP3 at protein level in the tumor tissues was significantly higher than that in the corresponding carcinoma adjacent tissues.The relative expresssion ULBP3 at mRNA level in carcinoma tissues of ESCC patients with lymph node metastasis and at TNM stage Ⅲ was higher than that of ESCC patients without metastasis and at TNM stage Ⅰand Ⅱ (t=4.839,4.192,P＜0.05).There was no significant correlation between the expression and ages,gender,location of tumors and the differentiation degree of tumor (P＞0.05).At early and mid stage of the tumor,the expression of ULBP3 at mRNA level was positively correlated with the percentage of NK cells in peripheral blood (r=0.5233,P＜0.05),however there was no correlation at advanced stage.Conclusion ULBP3 was highly expressed in ESCC and may be involved in the immune regulation of NK cells.

Patients with metastatic prostate cancer are typically managed with androgen deprivation therapy. Most patients initially respond to treatment, but many eventually develop castration-resistant prostate cancer. Neuroendocrine prostate cancer (NEPC) is a highly aggressive subtype of castration-resistant prostate cancer, which often results from neuroendocrine differentiation of prostate cancer cells. NEPC has a dismal outcome with an average survival of less than 1 year and exhibits less response to radiochemotherapy. At least 25%of patients with lethal castration-resistant prostate cancer are predicted to eventually develop this type of highly-aggres-sive NEPC. However, research on the molecular mechanism of NEPC is limited;thus, further studies are needed to explore the develop-ment and application of anticancer drugs for NEPC.

An HPLC method has been developed to determine polydatin in giant knotweed rhizome. In order to systematically validate the method, specificity, precision, linearity of reference solution and test solution, repeatability, reproducibility, accuracy, stability and robustness were measured. In the robustness test, a one-variable-at-a-time procedure was applied to evaluate the influence of slight variations in method factors, including the flow rate, the column temperature, the extraction time, and etc., on the assay result of polydatin. No significant differences were found when the process parameters changed during the experimental domain. And system suitability test limits were defined based on the robustness test. Results showed that the developed method was accurate, reproducible and robust.

Objective To investigate the site,MR signal types and morphological characteristics of ulnar nerve in cubital tunnel for healthy adults,in order to improve the awareness of ulnar nerve.Methods Unilateral elbow of forty healthy volunteers were scanned with MR,the sequences were as follows:T1-weighted-spin-echo,T2-weighted-spin-echo-fat-suppression,PD-weighted-spin-echo-fat-suppression,among 40,13 were supine,27 were prone.The site and MR signal types and morphological characteristics of ulnar nerve were observed,the long diameter and short diameter of the ulnar nerve on different axials were respectively measured.Results On axial,ulnar nerve was posterior to the medial condyle of the humerus at proximal elbow,lied between the flexor carpi ulnaris and the flexor digitorum profunds and superficialis muscles distally.The shape of the ulnar nerve was roundness or elliptic,the signal of 40 volunteers'(100％) ulnar nerve was isointensity on T1-weighted,the signal of 17 volunteers'(42.5％)ulnar nerve was isointense on T2-weighted or PD-weighted,the MRI signal of 23 volunteers' (57.5％) ulnar nerve was slight hyperintense on T2-weighted or PD-weighted,especially on the axial of medial condyle of humerus.The variation range of long diameter and short diameter of the ulnar nerve respectively were 1.4-3.8 mm,1.0-3.0 mm.Conclusion The certain location,MR signal types and morphological characteristics of normal ulnar nerve play an important role in the diagnosis of cubital tunnel syndrome.

Objective To study the expression of Gli1 protein and vascular endothelial growth factor (VEGF)-C in papillary thyroid carcinoma and their relationship with cervical lymph node metastasis.Methods The expression of Gli1 protein and VEGF-C were investigated by immunohistochemical EnVision method in 67 cases of papillary thyroid carcinoma and 32 cases of nodular goiter pathological specimen tissues.Nascent microvascular and micro-lymphatic of all pathological speimen tissues were examined by immunohistochemical monoclonal antibody CD34 and D2-40 staining assay respectively,and the microvascular density (MVD) and micro-lymphatic vessel density (MLVD) were calculated.The relationship between the positive expression of Gli1 protein and MVD,between the positive expression of VEGF-C and MLVD,and their relationship with cervical lymph node metastasis were analyzed.Results The positive rates of Gli1 protein,VEGF-C and MVD,MLVD were significantly higher in papillary thyroid carcinoma than those in nodular goiter [67.16％(45/67) vs.43.75％(14/32),70.15％(47/67) vs.31.25％(10/32),23.14 ± 2.06 vs.2.14 ± 0.31,13.36 ± 1.32 vs.3.53 ± 0.65,P ＜ 0.05].The positive expression of Gli1 protein was related to the patient' s age and the tumor size (P ＜ 0.05),while the positive expression of VEGF-C was not related to the patient' s age and the tumor size (P ＞ 0.05).The positive expression of Gli1 protein and VEGFC were significantly higher in TNM stage Ⅲ-Ⅳ of papillary thyroid carcinoma than those in TNM stage Ⅰ-Ⅱ (P ＜ 0.01) and also was significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.01).MVD and MLVD were significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.05).The positive expression of Gli1 protein was positively correlated with MVD (r =0.784,P＜ 0.01),the positive expression of VEGF-C was positively correlated with MLVD (r =0.529,P ＜ 0.01),the positive expression of Gli 1 protein was positively correlated with the positive expression of VEGF-C (r =0.586,P ＜0.01).Conclusions Gli1 protein which may participate in the formation of nascent microvascular is abnormally activated by the Hedgehog signaling pathway to express,VEGF-C which may be participate in the formation of nascent micro-lymphatic is mediated and started by the MAPK signaling pathways to express.Expressions of Gli1 protein is positively correlated with expressions of VEGF-C,therefore the Hedgehog signaling pathway may be associated with the MAPK signal pathway.Suppressing nascent microvascular and micro-lymphatic may become new target to blockingup papillary thyroid carcinoma cervical lymph node metastasis.

Objective To investigate the clinicopathologic features and imaging findings of several kinds of rare and uncommon brain embryonal tumours. Methods MRI or CT appearance and clinicopathologic features in 10 eases of histopathology confirmed brain embryonal tumours were retrospectively analyzed.Results Among 10 cases,there were supratentorial primitive neuroectodermal tumour(sPNET) in 6,ependymoblastoma in 2,medulloepithelioma and atypical teratoid/rhabdoid tumour (AT/RT)in 1 respectively.Patients in 4 cases were infancy and childhood.There was characteristic of the clinicopathologic features of tumors that were the evidence of accurate diagnosis. All lesions were located in the supratentorial region.The tumors were large,usually lobulated in shape irregularly and showed heterogeneous density or signal intensity with some cystic and necrotic areas.The tumors parenchyma appeared slightly high density on CT imaging,iso-or high signal on T2WI and slight low signal on T1WI with intense enhancement after contrast injection.Conclusion MRI or CT findings of rare and uncommon brain embryonal tumours were not characteristic,it Was very essential to know the tumors combined with the clinical and pathologic features.

micmRNA (miRNA) plays an important role in biological processes via regulating gene expression in the level of posttranscription.Increasing number of studies reveal that miR-375 is downregulated in digestive system tumors, and is related to tumorigenesis.Overexpression of miR-375 can inhibit growth and metastasis of tumors through repressing the expression of target genes.

Objective To analyse a novel splice mutation in EXT1 gene of hereditary multiple osteochondroma, and study its pathogenic mechanism.Methods In April of 2013, the proband was hospitalized from the outpatient department with multiple joint deformity for more than 20 years, peripheral blood of the proband and his parents were collected and genomic DNA was extracted .Coding regions and adjacent intron sequences of EXT1/EXT2 genes in genomic DNA of the family members were amplified and sequenced.Bioinformatics was used to analyze the mutation from sequencing .cDNA from peripheral blood of the proband ,the mother and normal control was made respectively as a template for amplifying coding regions of EXT1 gene, and the product was T-A cloned and sequenced.The abnormal transcripts of each group were counted and analyzed using chi square test to study the pathogenic mechanism of the mutation .Results Sequencing results of family members revealed that there was a heterozygous deletion mutation ( c.1284 +2del) in the 5′splice site of intron 4 in EXT1 gene of the proband and his mother .Bioinformatics predicted that exon 4 of EXT1 gene was skipping or spliced aberrantly due to the mutation .T-A clone and sequencing results as well as the statistical analysis suggested that there was a significantly higher proportion of transcripts with skipping exon 4 in the proband and his mother compared with the normal control (P=0.000, P<0.01).Conclusions c.1284+2del in EXT1 gene is reported for the first time internationally , which results in a considerable number of abnormal transcripts with skipping exon 4 in EXT1 gene, thereby influences the normal transcription and translation of EXT1 gene.

ObjectiveTo observe the effect ofTong Du acupuncture-moxibustion, conventional acupuncture, and medication on the bone marrow stem cells (BMSCs) mobilization markers CD34+and CD133+, neurological deficit score in patients with ischemic stroke.Method Totally 120 eligible subjects with ischemic stroke were randomized into aTong Du acupuncture-moxibustion group (group A), a conventional acupuncture group (group B), and a medication group (group C), 40 in each group. In addition to the conventional medication given to all three groups, patients in group A also received moxibustion at Baihui (GV20), bloodletting at Dazhui (GV14), and acupuncture at Fengchi (GB20), Quchi (LI11), Hegu (LI4), Zusanli (ST36), and Sanyinjiao (SP6); patients in group B also received acupuncture at Fengchi, Quchi, Hegu, Zusanli, and Sanyinjiao; group C received patent Chinese medicine Tong Xin Luo capsules. The intervention lasted 14 d in all three groups. The CD34+and CD133+and neurological deficit score were evaluated and compared before and after intervention.Result The three treatment methods all significantly increased the level of CD34 + and CD133+in peripheral blood (P<0.05,P<0.01); the intergroup comparisons showed significant differences among the three groups (P<0.05), and the level in group A was significantly different from that in group B and C (P<0.05), while there was no significant difference between group B and C (P>0.05); the three treatment methods all improved the neurological deficit score to various extent, and the result in group A was significantly different from that in group B and C (P<0.05,P<0.01), and there was also a significant difference between group B and C (P<0.05).ConclusionsTong Du acupuncture-moxibustion, conventional acupuncture, and medication can influence the BMSCs and neurological deficit to different extent; the effect ofTong Du acupuncture-moxibustion on the BMSCs and neurological deficit is more significant than that of the conventional acupuncture and medication, suggesting thatTong Du acupuncture-moxibustion method should have advantage in treating cerebral infarction.