pPIC9 was used as a template and ?-factor gene of Saccharomyces cerevisiae was amplified by PCR.The gene was cloned in the intracellular expression vector pYES2/CT for Saccharomyces cerevisiae and the secreting expression vector named pYES2/CT/?-factor(pYC?)was constructed.The gene of mannase(man)from recombinant vector of pKLAC1-man(pKLman) was cut by restriction enzymes and linked with pYC?.This recombinant vector pYC?-man was used to determine the secretory ability and stability of pYC?.The excellent secretory ability of pYC? was proved by two experiments.One showed that INVSc1/pYC?-man clones formed the clear rings around the clones on the medium contained trypan blue,while INVSc1/pYC? clones had no rings.Further analysis of mannase activity of extracellular supernatant and intracellular extracts showed that both extracellular and intracellular mannase activities of INVSc1/pYC? were not detected,while INVSc1/pYC?-man had evident extracellular mannase activities and no intracellular mannase activities.The stability of pYC? was also very good proved by continuous cultivation for about 150h.

To investigate the effect of preceding naloxone injection into the third cerebroventricle or acute subdiaphragmatic vagotomy on the gastric acid secretion inhibited by the somatostatin analogue octreotide given by intracerebroventricular (icv) injection. The third ventricles were cannulated in male Wistar rats anesthetized with sodium pentobarbital. One week later, acute gastric lumen perfusion was carried out. The gastric perfusion samples were collected every 10 min and were titrated by 0.01 mol/L NaOH to neuter. On the basis of subcutaneous injection of pentagastrin (G-5, 160 micro, g/kg), icv injection of physiological saline (group A, n = 20), icv injection of octreotide (0.05 micro g) (group B, n = 20), icv injection of naloxone (2.5 micro g)+octreotide (0.05 micro g) (group C, n = 20), acute subdiaphragmatic vagotomy+ icv injection of physiological saline (group D, n = 20), or acute subdiaphragmatic vagotomy+icv injection of octreotide (0.05 micro g) (group E, n = 20) were conducted. Before and after icv injection, 1-h total acid output (TAO) was determined and compared. The experimental data were expressed in change rate (%) of TAO. The change rates (%) of TAO were 4.60% in group A, -20.35% in group B, -18.06% in group C, 5.01% in group D and -21.59% in group E, respectively. Comparison of group B or C versus group A showed that P < 0.01 and comparison between the group E versus group D showed that P < 0.01. Whereas the differences between group C and group B, group E and group B were not statistically significant (P > 0.05 for all). The results indicate that the central inhibition of gastric acid secretion by octreotide may not be mediated by the endogenous opiate substance or its receptor and the peripheral pathway for icv injection of octreotide to suppress gastric acid secretion is via extra-vagus route.

Objective To improve the unsatisfactory situation of endoscope disinfection and develop endoscopic anti-cross- infection security protection sheath. Methods The protective sheath was processed by using tube and cylinder of medical flexible rubber that was non -toxic, harmless, tasteless and transparent, which size was coincide with the shank of endoscopic fore-end. The openings of fore-end and back-end were designed by immerse extra-strong circle. Every sheath was molded into barrel roll based on the midline axis of twist and bagging standby after disinfection and sterilization. Results The instruction of sheath was streamlined that could be unwound smoothly from back-end and accreted tightly to endoscopic mirror. Conclusion The sheath can be reduced effectively the risk of cross-infection in endoscope examination.

Objective To evaluate modified retroperitoneal laparoscopic resection of renal cyst. Method Thirty-six patients with renal cyst were treated by modified retroperitoneal laparoscopic resection of renal cyst,summarized the clinic data and follow-up the effect. Results All 36 cases were operated suc-cessfuUy without changing to opening operation,average operation time (50 ± 35)min,no complications oc-curred and no recurrence was found. Conclusions The modified retroperitoneal laparoscopic resection of renal cyst with two 5 mm-trocars and one 10 mm-trocar has less trauma than classic laparoscopic operation. It is one of mini-trauma operation method which is worth to be popularized in clinic.

Objective:To assess the application value of reflectance confocal microscopy (RCM) in evaluating clinical efficacy of hemoporfin-mediated photodynamic therapy for purple-type port-wine stain.Methods:From April 2018 to January 2020, a total of 39 patients with centrofacial purple-type port-wine stains were enrolled from Department of Dermatology, Wuhan No.1 Hospital, and received 3 sessions of hemoporfin-mediated photodynamic therapy. Before the first treatment, and 3- 6 months after 3 sessions of hemoporfin-mediated photodynamic therapy, skin lesions were photographed, and RCM was conducted to measure the diameter and density of blood vessels at a depth of 100 μm in the lesions. Clinical efficacy was evaluated based on the clinical photos, and the average diameter of blood vessels and density of blood vessels per square millimeter of lesion area were calculated. Measurement data were compared among different groups by using one-way analysis of variance, multiple comparisons were performed using least significant difference test, and comparisons of parameters before and after treatment were conducted by using paired t test. Results:After 3 sessions of hemoporfin-mediated photodynamic therapy, 1 (2.56%) patient was nearly completely cured, 16 (41.03%) received marked improvement, 20 (51.28%) received improvement, and 2 (5.13%) showed no response to the treatment. In the patients receiving marked improvement or improvement, the average diameter and density of blood vessels significantly decreased after treatment compared with those before treatment (all P < 0.05) , while no significant difference was observed before and after treatment in the patients with no response (both P > 0.05) . The average difference in the blood vessel diameter before and after treatment was significantly higher in the patients receiving marked improvement (48.56 ± 17.87 μm) than in those receiving improvement (31.15 ± 21.09 μm, P < 0.05) and those with no response (12.00 ± 2.83 μm, P < 0.05) . The average difference in the blood vessel density before and after treatment was 7.13 ± 3.44, 5.00 ± 2.22 and -0.50 ± 3.54 vessels/mm 2, respectively, in the patients receiving marked improvement, improvement and those with no response, and pairwise comparisons between the 3 groups all showed significant differences (all P < 0.05) . Conclusion:RCM can be used to assess the average diameter and density of blood vessels in the port-wine stain lesions before and after hemoporfin-mediated photodynamic therapy, and is helpful in quantitatively evaluating the therapeutic effect of hemoporfin-mediated photodynamic therapy.

Objective To evaluate 10-year risk of ischemic cardiovascular diseases (ICVD) in middle-aged adults in Henan province to provide evidence for improved health status. Methods The 10- year risk of ICVD in 12 064 middle-aged adults was evaluated by using simplified risk estimation model. Results About 96. 35% of male and 98. 29% of female had a probability of < 10% to develop ICVD over 10 years; 3.65% of male and 1.71% of female had a probability of ≥ 10% to develop ICVD over 10 years; and 0.68% of male and 0.23% of female had a probability of≥20% to develop ICVD over 10 years. The prevalence of risk factors for ICVD in two absolute risk groups ( ≥10% vs < 10% ) was significantly different. Conclusions The risk of ICVD in the middle-aged residents of Henan province may be similar to that in the middle-aged Chinese adults. The risk assessment for ICVD could be used for high- risk populations.

Objective To compare the effect and safety of arthroscopic debridement enmbined with intercondylar fossa angioplasty in the elder with knee osteoarthritis.Methods Six-eight elderly with knee osteoarthritis were selected from June 2012 to December 2014 in this hospital and they were averagely and randomly divided into group A and B.Patients in group A were given arthroscopic debridement,while in group B were given and arthroscopic debridement,intercondylar fossa angioplasty.The levels of VAS scores,Lysholm scores,WOMAC scores and joint range of motion were measured and compared between the two groups after 3 months,6 months and 12 months of operation.The adverse events of the two groups were supervised and compared as well.Results All elder in the two groups had apparently decreasing in VAS scores and WOMAC scores (P＜0.05) after operation.Compared with group A,patients in group B had lower levels of VAS scores and WOMAC scores(P＜0.05) at 12 months after operation.The level of Lysholm score was significantly increased in the two groups after treatment(P＜0.05),while group B had a higher level at 12 months after treatment than group A (P＜0.05).The joint range of motion in the two groups were improved after treatment(P＜0.05).The elder patients in group B had better joint range of motion than group A at 6 and 12 months after operation (P＜0.05).Early postoperative swelling in the joints was the adverse event after operation,but there was no statistically difference between two groups.Conclusion The combination of arthroscopic debridement and intercondylar fossa angioplasty is effective to release knee pain,increase the clinical efficacy and improve the knee function.

Objective: To investigate the effects of different extracts of Smilax china L on the activity of ovarian cancer cells. Methods:Solvent extraction method was used to extract the active ingredients of Smilax china L. , and CCK-8 assay method was ap-plied to detect the influence of different Smilax china L. extracts (0, 50, 100, 150 and 200μg·ml-1 ) on the survival rate of ovarian cancer cells including low invasiveness A2780 cells and high invasiveness HO-8910PM cells. At the same time, the status of the two kinds of ovarian cancer cells at different time points (24, 48 and 72 h) was observed. Results:The IC50 of N-butanol extracts (SCR-B) on HO-8910 and A2780 ovarian cancer cells was 47. 5 μg· ml-1 and 69. 2 μg· ml-1 , respectively, and that of ethyl acetate ex-tracts (SCR-E) on A2780 and HO-8910 cells was 147. 9 μg· ml-1 and 166. 0 μg· ml-1, respectively. Smilax china L. extracts had the inhibition against both A278 and HO-8910PM ovarian cells in a dose-and time-dependent manner. Conclusion:The inhibitory activity of SCR-B against ovarian cancer cells is stronger than that of SCR-E, and SCR-B has stronger inhibition against A2780 cells than against HO-8910 cells. SCR-B has better inhibition against ovarian cancer cells.

Intraductal papillary mucinous neoplasm (IPMN) is a kind of epithelial neoplasms of mucin-producing cells arising in the main duct and/or branch ducts of the pancreas,and it is one of the precancerous lesions of the pancreatic cancer.With the development of pathology and radiology,the diagnostic rate of IPMN has been gradually increased and given a new understanding on the pathological characteristics,clinical diagnosis and treatment of IPMN.This review overviewed the classification,diagnosis,management and prognosis of IPMN,aiming to deepen the understanding of IPMN and improve the level of diagnosis and treatment.

Objective:To investigate the clinical features and adjuvant chemotherapy of children with medulloblastoma(MB).Methods:Clinical data of 21 pathologically confirmed MB children admitted to the department of pediatrics of Beijing Tongren Hospital affiliated to Capital Medical University from May 2012 to November 2017 were collected to analyze the clinical efficacy and prognosis of multidisciplinary combined treatment.Results:There were 21 children enrolled in the study(15 males and 6 females; median age: 6 years and 3 months). The majority of tumors were from the fourth ventricle(66.7%, 14/21 cases). The most common type of pathological tissue was classic medulloblastoma(61.9%, 13/21 cases). Most of the molecular types was type 4(47.6%, 10/21 cases). There were 15 cases(71.4%)in the high-risk group and the remaining 6 cases(28.6%)in the low-risk group without metastasis(M0 stage). Total tumor resection was performed in 16 cases(76.2%). The patients were followed up to December 2019(median follow-up time was 29 months). After comprehensive treatment, 11 patients died and 6 patients relapsed.The 2-year survival rate was 61.5%, and the 5-year survival rate was 51.1%.Cox regression multivariate analysis showed that the survival rate of children with no tumor spread, short time interval between radiotherapy and surgery was higher( P<0.05). Conclusion:The incidence of MB in boys is higher than that in girls.Whether the tumor is disseminated or not, the time interval between radiotherapy and surgery are independent risk factor affecting the prognosis.Multidisciplinary combination therapy can effectively improve the long-term prognosis.