This study was aimed to explore a new method to identify the original plant of Plantaginis Semen and its adulterants by the ITS2 regions. The second internal transcribed spacer (ITS2) of ribosomal DNA was amplified and sequenced by bidirectional sequencing of PCR products. Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner. The ITS2 secondary structure was predicted using ITS2 database and web-sites. The phylogenetic tree was constructed by MEGA5. The results showed that the maximum intraspecific K2P dis-tance of Plantago asiatica was 0.009 9, while the minimum interspecific K2P distance was 0.497 6; the maximum in-traspecific K2P distance of P. depressa was 0.005 2, while the minimum interspecific K2P distance was 0.519 1. The ITS2 secondary structure showed that P. asiatica and P. depressa can be differentiated obviously from its adulterants. Different samples of P. asiatica and P. depressa were gathered together and can be distinguished from its adulterants by NJ tree. It was concluded that the ITS2 sequence was able to identify original plant of Plantaginis Semen and its adulterants correctly. It provided a new method for the identification of original plant of Plantaginis Semen.

Objective To investigate the relationship of PI3K/Akt signaling pathway with the activation of hepatic stellate cells (HSC) and its role in radiation-induced hepatic fibrosis.Methods HSC was treated with 6 MV X-ray irradiation (IR) together with the inhibitor of PI3K/Akt signaling pathway.The cells were divided into inhibitor group,10 Gy IR group,10 Gy + inhibitor group,20 Gy IR group,an 20 Gy + inhibitor group and blank control group.Then cell apoptosis rate was detected,the expression of transforming growth factor β1 (TGF-β1) in cell supernatant and the mRNA expressions of α-smooth muscle actin (α-SMA) and phosphorylation protein kinase B (p-Akt) were measured.Results Compared with the control group,the apoptosis rate of 10 and 20 Gy IR group increased with irradiation dose (t =8.43,11.63,P ＜0.05) but they were reduced by the inhibitor of PI3K/Akt (t =8.09,4.88,P ＜0.05).The expressions of TGF-β1,α-SMA,and p-Akt also increased with irradiation dose (t =6.91,7.80,9.28,P＜0.05) but they were declined by this inhibitor for both 10 Gy IR (t =6.17,15.11,10.34,P＜0.05) and 20 Gy IR (t =10.04,6.85,23.84,P＜0.05).Conclusions X-ray irradiation could activate HSC through PI3K/Akt signaling pathway,which may further result in hepatic fibrosis.

Objective To analyze the clinical epidemiology of primary hepatocellular carcinoma (PHC) in the Xinjiang region. Methods Clinical data of the patients with PHC were collected at First Affiliated Hospital of Xinjiang Medical University from 5 577 cases from January 2002 to December 2014, their gender, race, age, household distribution, hepatitis virus-positive rate were analyzed retrospectively. Results Among the 5 577 eases, the men/women gender ratio was 3.45∶1;the proportion of Han, Uighur, Kazakh, and other ethnic groups (Hui, Mongolian, Manchu, Xibo) was 79.67%, 9.86%, 4.55%, 3.31%and 2.61%, respectively. The Constituent ratio difference between Uighur and Han was significant (P<0.05);4 232 patients had hepatitis B surface antigen (HBsAg) detection, and 3 833 patients had HCV antibody (HCV-Ab) detection. HBsAg was positive in 2 560 cases (60.49%), HCV-Ab was positive in 490 cases (12.78%). Hepatitis B virus detection positive rate in Uygur was 35.52%, in Kazak was 40.00%, which was lower than the Han's (65.68%, P<0.05). Urban and rural population had 3589 cases (64.35%) and 1988 cases (35.65%). Conclusion An increased risk for PHC was found in hepatitis virus-positive patients, the Xinjiang Uygur and Kazak people had significantly lower prevalence of HBV infection than the Han's. Appropriate measures should be taken for clinical diagnosis, treatment and prevention of PHC.

Objective:To establish the ISSR fingerprint of Chrysanthemum morifolium cultivated in Futianhe area of Macheng county to guide the breeding of C. morifolium. Methods:Using the technology of ISSR molecular markers and the software of SPSS 15. 0, the coefficient matrix of Jaccard was established and the tree graph of the genetic relationship of the breeds of C. morifolium cultivated in Fu-tianhe area was built to analyze the respective genetic relationship and features. Results:By ISSR analysis, it confirmed that C. morifoli-um in Futianhe area had long genetic distance with the other white chrysanthemum breeds, and it could be considered as an individual breed. Conclusion:The ISSR map can be used to identify the breed of C. morifolium cultivated in Futianhe area.

Objective To investigate the feasibility and efficiency of unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion to treat low lumbar vertebra diseases. Methods Thirty patients with low lumbar vertebra diseases were entered into the study, including 8 males and 22 females with an average age of 53.7 years. All patients underwent discectomy, spinal canal decompression, cage implantation and lumbar fixation by unilateral pedicle screw combined with contralateral translaminar facet screw under gunsight guiding by percutaneous. Clinical outcomes were assed by JOA questionnaires before and after operation. Operative time, blood loss, and postoperative draiming were recorded. Radiological examination was obtained to assess position of translaminar facet screw.Results Mean operation time was 89 min with a blood loss of 285 ml. Position of translaminar facet screw grade Ⅰ were 24 cases, and grade 11 were 6. Mean follow-up was 22.5 months. 29 cases got bony fusion, and the fusion rate was 96.7%. There were no instability and evidence instrument failure during follow-up. The JOA grades improved from 13.0 preoperation to 25.2 at final follow-up, with the excellent and good rate of 72.5 %. Conclusion Unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion provide simple procedure, little trauma, forceful fixation, high fusion rate, and less complication, etc. Therefore, the surgical maneuver is a good choice for partial low lumbar vertebra diseases.

UL48 plays essential role in replication of MDV genome and interacts with UL36 as well as other MDV tegument proteins.To investigate the interaction between UL48 and UL36 during MDV oncogenisis,antibody against UL48 was prepared and characterized in current study.UL48 gene was amplified from MDV-Ⅰ genome and then subcloned into pTYB1 and pGEX-4T3 vectors for UL48 expression with induction of IPTG in BL21(DE3) E..coli cells.Chitin-sepharose and Glutathion-sepharose were,respectively,used to purify fusion protein intein-UL48 and GST-UL48.Four subcutaneous injections of intein-UL48 fusion protein were done on the lower back and the thigh of rabbit and then other three injections with an interval 10 days.The titer of antibody was measured by the sandwich ELISA with UL48 protein isolated from GST-UL48 after cleavage of thrombin.Western blot was carried out for specificity analysis of antibody against UL48 protein.The results suggested that UL48 antibody was succesfully prepared,and its titer was 1 ∶ 512 000.

Objective To analyze the clinical and molecular genetic characterizations of X-linked adrenal dysplasia congenita(AHC) onset in infant.Methods Seven children (from 7 families) with X-linked AHC who were admitted to the Department of Endocrinology,Genetics and Metabolism,Children's Hospital Affiliated to Zhengzhou University,from July 2012 to June 2017 were selected.All patients were screened for dosage-sensitive-sex reversal-adrenal hypoplasia congenital critical region on the X chromosome gene1 (DAX1/NR0B1) mutations.The clinical manifestation and laboratory examination were analyzed,their clinical characterizations were summarized.Results Seven patients were all male,the onset age of the patients were from after birth to 7 months old,and 4 patients (4 families) had a family history of X-linked recessive inheritance.The clinical manifestations were skin pigmentation [100.0％ (7/7 cases)],vomiting [71.4％ (5/7 cases)],no weight gain [57.1％ (4/7 cases)] and poor spirit [28.6％ (2/7 cases)].Laboratory tests showed that hyperkalemia and hyponatremia,increased coricotrophin,normal or decreased cortisol,17α-hydroxyprogesterone,progesterone,aldosterone and dehydroepiandrosterone.Testosterone levels increased in 5 patients.The abnormalities of adrenal glands imaging could be seen in 2 patients.Two patients were misdiagnosed as congenital adrenal cortical hyperplasia.Then the definitive diagnosis were made by genetic test.DAX1/ NR0B1 gene mutations were found in all patients.Five patients were novel mutations (c.114_126del,c.872G ＞ A,c.56delG,c.884T ＞ G,c.1217delG).Conclusions The clinical manifestations of X-linked AHC with infant onset include pigmentation,poor spirit and growth retardation,which should be differentiated from congenital adrenal cortical hyperplasia.Hormone levels such as elevated blood 17α-hydroxyprogesterone and family history are the main identification points,and AHC cannot be excluded when testosterone level increases.Five novel mutations are found in this study,which enrich the gene database.

Objective To investigate the Th1/Th2 cytokines and chemokine CXCL 13 levels in peripheral blood and cerebrospinal fluid(CSF)of patients with neurosyphilis and their clinical significance. Methods Forty seven HIV negative patients with neurosyphilis, 36 syphilis patients without neurological involvement(syphilis group)and 23 patients without infectious intracranial diseases(control group) admitted in Hangzhou Third Hospital during July 2011 to December 2014 were enrolled in the study.CSF pressure, protein contents, white blood cell counts and IgG index were detected in patients with neurosyphilis.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of Th1 cytokines (IL-2,IL-12 and IFN-γ),Th2 cytokines(IL-6,IL-10)and CXCL13 in serum and CSF in three groups.Th1/Th2 cytokines,CXCL13 levels and CSF routine were also examined in neurosyphilis patients after treatment for 3 and 12 months.ANOVA, Kruskal-Wallis test and Spearman correlation were used for data analysis.Results The levels of IL-6 and IL-10 in serum and CSF from neurosyphilis patients were higher than those in syphilis group and control group(χ2IL-6=15.43, 15.39 and 14.44, 20.01,χ2 IL-10 =16.46, 23.86 and 15.11,24.44;P<0.05 or <0.01), while the levels of IL-2, IL-12 and IFN-γin the serum and CSF were lower than those in syphilis group and control group(χ2IL-2=14.55,17.14 and 16.14,17.97;χ2IL-12=13.65,20.50 and 18.48,21.04;χ2IFN-γ=16.95,17.53 and 16.00,15.21;P<0.05 or <0.01). CXCL13 contents in CSF of neurosyphilis patients were significantly higher than those in other two groups (χ2=52.51 and 53.76, P <0.01), and were positively correlated with leucocyte counts, protein concentrations,IgG index,IL-6 and IL-10(r=0.325,0.544,0.750,0.333 and 0.382,P<0.05 or <0.01),but were negatively correlated with IL-12 levels in neurosyphilis patients(r=-0.303,P<0.05). In neurosyphilis patients,CXCL13 and IL-6 levels were reduced after treatment for 12 months(χ2=102.00 and 22.17,P<0.05 and <0.01), while the levels of IL-2 and IL-12 were increased(χ2=18.28 and 24.10,P<0.05 and <0.01).Conclusion Chemokine CXCL13 and Th1/Th2 cytokines are involved in the immune response in neurosyphilis patients, which may be used for the diagnosis and evaluation of therapeutic efficacy in patients with neurosyphilis.

Objective To examine the feasibility and early effects of unilateral pedicle screw fixation in combination with contralateral translaminar facet screw fixation plus interbody fusion through the channel-assisted muscle-splitting approach for the treatment of degenerative lumbar spine diseases in elderly patients.Methods A total of 82 elderly patients with degenerative lumbar spine diseases received treatment between January 2015 and June 2016.Of these patients,25 had lumbar spinal stenosis,42 had degenerative lumbar instability,13 had giant lumbar disc herniation,and two had recurrent lumbar disc herniation;There were 69 cases of single segment and 13 cases of double segment lesions.Perioperative evaluation,imaging evaluation,and clinical evaluation were performed,and surgical complications were analyzed.Results The average length of incision was (2.6 ± 0.4) cm in cases of single segment lesions and (4.4±0.5) cm in cases of two segment lesions.The mean operation time was (98.3±19.6) minutes and peri-operation blood loss was (232.2±25.8) ml.The post-operation height of the intervertebral disc space significantly increased at the last follow-up(P ＜ 0.05);the height of the intervertebral disc space was restored and well maintained with internal fixation showing no loosening,breakage or Cage shift.The post-operation lumbar coronal Cobb angle decreased(P＜0.05)while the sagittal Cobb angle increased(P＜0.05)at the last follow-up.There was no significant difference in the area and grade of the multifidus muscle on MRI 12 months after operation.Seventy-two patients were followed up for 12-18 months (mean =16.8 months),and there were 67 cases of intervertebral fusion (93.1％).The last follow-up showed a Japanese Orthopaedic Association(JOA) score of (25.8 ± 1.7),which was significantly higher than the pre-operation score (10.6±1.3),with an excellence rate of 88.9 ％.The Oswestry Disability Index score was significantly reduced,from (50.9±21.6) at pre-operation to (9.6± 4.8) at the last follow-up.There were 78 patients with primary wound healing,three cases of superficial wound necrosis,one case of dural tear,and one case of nerve root injury.No cardiovascular or cerebrovascular incident was observed during or after operation.Conclusions Combined pedicle/facet screw fixation plus interbody fusion through the channel-assisted muscle-splitting approach shows satisfactory short-term effects when used for the treatment of degenerative lumbar spine diseases in the elderly and possesses advantages such as minimal invasiveness,limited bleeding,quick recovery,high fusion rates and significantly improved clinical outcomes.However,further clinical studies are required to assess its medium-and long-term efficacy.

Objective To study the protective effect of shenfu injection on cerebral ischemia reperfusion injury in rats.Methods 120 male Sprague Dawley (SD) rats(320-350 g) were randomly divided into sham operation group,model control group,Nimodipine injection group,low,medium and high dose group of shenfu injection according to gender weight.20 males in each group were given medicine once a day for 7 days before operation.The cerebral ischemia model was established by thread embolization after 5 days of administration.In the sham operation group,the other operations were the same as those in the model group except for carotid artery ligation and thread insertion.After 24 hours of perfusion,the neurological score,abdominal aorta blood flow,malondialdehyde (MDA),superoxide dismutase (SOD) and glutathione (GHS) levels in brain tissues were measured.Triphenyltetrazolium chloride (TTC) staining was used to calculate the area of cerebral infarction and pathological examination of brain tissues.Results Compared with the model control group,the middle and high dosage of shenfu injection could obviously improve the nerve function and increase the percentage of cerebral infarction area (P ＜ 0.05);the high dosage group of shenfu injection could obviously decrease the whole blood viscosity (P ＜ 0.01);the middle and high dosage of shenfu injection could obviously reduce the level of MDA in rat brain tissue (P ＜ 0.01) while increasing the levels of SOD and GSH (P ＜0.01),finally could significantly improve the pathological changes of brain tissues such as mild swelling of nerve fibers,mild neuronal degeneration,inflammatory interstitial edema and inflammation.Conclusions Shenfu injection has obvious protective effect on cerebral ischemia reperfusion model in rats.