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 killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.

OBJECTIVE: To analyze the clinical and genetic characteristics of a patient with dihydrolipoamide dehydrogenase deficiency. METHODS: Potential variants of the DLD gene were detected by whole exome sequencing and verified by Sanger sequencing. RESULTS: Compound heterozygous variants, c.704_705delTT (p.Leu235Argfs*8) and c.1058T>C (p.Ile353Thr), were detected in the DLD gene. The c.1058T>C (p.Ile353Thr) variant was derived from his mother and known to be pathogenic. The c.704_705delTT (p.Leu235Argfs*8) variant was derived from his father and was unreported previously. CONCLUSION The compound heterozygous variants of c.704_705delTT (p.Leu235Argfs*8) and c.1058T>C (p.Ile353Thr) of the DLD gene probably underlay the disease in this patient. Above finding has facilitated genetic counseling and prenatal diagnosis for the family.
Acidosis, Lactic/genetics* ; Dihydrolipoamide Dehydrogenase/genetics* ; Female ; Genetic Testing ; Genetic Variation ; Humans ; Male ; Maple Syrup Urine Disease/genetics* ; Pregnancy ; Whole Exome Sequencing

OBJECTIVE: To analyze the clinical and genetic characteristics of a patient with congenital isolated adrenocorticotropic hormone deficiency (IAD). METHODS: Clinical characteristics of the patient was reviewed. Genomic DNA of the child was subjected to whole exome sequencing. RESULTS: Genetic testing has confirmed the diagnosis of congenital IAD by identification of compound heterozygous variants of the TBX19 gene, which included a pathogenic nonsense c.535C>T (p.R179X) variant inherited from his father and a novel missense c.298C>T (p.R100C) variant inherited from his mother. CONCLUSION Congenital IAD due to variants of the TBX19 gene is a rare autosomal recessive disease. It is characterized by low plasma adrenocorticotropic hormone and cortisol levels but normal levels of other pituitary hormones. Delayed diagnosis may lead to severe early-onset adrenal failure and wrong treatment which may result in neonatal mortality. Hydrocortisone replacement is effective. Detection of pathogenic variant of TBX19 gene is the key to diagnosis.
Adrenal Insufficiency/genetics* ; Child ; Homeodomain Proteins/genetics* ; Humans ; T-Box Domain Proteins/genetics*