Objective Artificial mineral water pills were made by loading many kinds of trace elements into the ceramic carrier.Pure water could be mineralized after dipping the pills into the water.To assess the possible hazardous effects on the human health induced by the pills,a series of hygienic and toxicological experiments on the pills were carried out.Methods Acute oral toxicity test,micronucleus test of mice bone marrow cells,sperm abnormality test in mice and Ames test were con-ducted.Contents of trace elements,heavy metals and radioactivity in the mineralized water were determined according to the examination methods in Standard Examination Methods for Drinking Water and Examination Methods for Drinking Natural Min-eral Water.Results The contents of trace elements,heavy metals and radioactivity in mineralized water met the requirements of GB5749-1985Sanitary Standard for Drinking Water and GB8537-1995Sanitary Standard for Drinking Natural Mineral Wa-ter.Mineral pills were classified as actually non-toxic and the results of micronucleus test of mice bone marrow cells,sperm ab-normality test in mice and Ames test were negative.Conclusion Mineralized drinking water treated by artificial mineral pills were safe.

Objective: To investigate the preparation of pulsatile release tablets, the release of the drug in vitro and the pharmacokinetics in vivo . Methods: Dil tiazem hydrochloride（DIL） was used as model drug. The pulsatile release tabl e ts were prepared by dry-coated method with carnauba wax, bee wax and hydrophil i c cellulose as coating materials. The effects of formulation and technology on t he release characteri stic of diltiazem hydrochloride was investigated. The mechanism of pulsatile rel ease of the drug was proved by erosion test. The pharmacokinetic study on four h uman subjects was done by means of HPLC measurement. Results: In vitro ， delayed-release ti me t 10 was 2.1　h, the maximum release time t rm 4.0　h and t he pulsed-releas e time t 10-90 1.7　h. In vivo ， delayed-release time t la g was 5.7　h, the p eak time 8.5　h and the pulsed-release time 2.6　h. Conclusion: The rele ase of drug from pulsatile-released tablets of diltiazem hydrochloride was in a pulsed way both in vitro and in vivo .

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.

Objective To explore the dynamic changes of serum S100B and glial fibrillary acidic protein (GFAP) levels and their clinical significance in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).Methods By means of enzyme-linked immunno-sorbent assay (ELISA),the serum S100B and GFAP levels from 33 DEACMP patients were assayed,and the condition changes were analyzed with three types of scale:the activity of daily living scale ( ADL),information-memory-concentration test (IMCT) and Hasegawa' s dementia scale(HDS).The comparison with 32 patients of acute carbon monoxide poisoning without DEACMP was also conducted.Results (1) The serum S100B( (0.60 ±0.21)ng/ml) and GFAP( (226.58 ±90.05 )ng/ml) in DEACMP group at acute stage were significantly higher than those in acute-CO-poisoning group ( (0.50 ± 0.20) ng/ml,( 183.04 ± 73.01 ) ng/ml) and those in DEACMP group at convalescent stage ( (0.51 ±0.16) ng/ml,(183.25 ±81.76)ng/ml) (all P values ＜0.05).(2)In DEACMP group,the serum S100B and GFAP at acute and convalescent stages were significantly correlated (at acute stage:r=0.466,P=0.006; at convalescent stage:r=0.365,P=0.037 ).(3)The S100B and GFAP in ineffective DEACMP patients at acute stage were significantly higher than those in the other groups ( all P values ＜ 0.05 ).(4) In DEACMP group,the ADL,HDS and IMCT scores( (45.21 ± 9.69),(8.26 ± 6.31 ),(9.91 ± 7.52) ) at acute stage were significantly different from those at convalescent stages( (33.67 ± 13.62),( 15.91 ± 10.83),( 19.06 ± 10.37 ) ) ( all P values ＜0.01 ).Conclusion There is secondary brain insult (SBI) in DEACMP; glial activation may play an important role.The S100B and GFAP levels may be associated with the prognosis of DEACMP.

Objective To observe the effect of propofol on cognitive functions and hippocampus tissue in hyperlipidemia aged rats.Methods Seventy-six aged male SD rats were selected and randomly divided into basal diet with propofol injection (group PI), basal diet with normal saline injection(group N1), high-fat diet with propofol injection (group P2) and high-fat diet with normal saline injection (group P2), with 19 rats in each group.Eight weeks later,group P1 and P2 received intraperitoneal injection of propofol 100 mg · kg-1 · d-1 for 5 days.While in group N1 and N2, rats received the same intraperitoneal injection of equal volume normal saline.One day after the last injection, escape latency and space exploration were detected by Morris water maze in the next six days.One hour after the last water maze test, the serum and hippocampus were sampled to detect the expression of beta-amyloid protein and receptor for advanced glycation endproducts(RAGE) by immunohistochemical method and ELISA respectively.Results In place navigation tests,the escape incubation period(98.20±25.40)s in group P1 was obviously longer than that in N 1 group (47.50± 11.08) s (P＜ 0.01).Compared with P2 (99.79 ± 20.38) s, escape incubation period was shortened in N2 (50.70± 20.55) s (P＜ 0.05).There was no significantly difference between N2 and N1 (P＞0.05), while the escape incubation period in group P2 was longer than P1 (P＜ 0.05).In spatial probe test,platform passing number ((2.86 ±1.46)times) in group P1 were less than that in N1 group ((7.50± 1.70) times, P＜0.05).In group N2, the platform passing number ((6.60 ±3.91) times) were more than those in group P2 ((1.16 ±1.16)times, P＜0.05) while there was no significant difference between group N1 and N2 (P＞0.05).Times of crossing platform in group P2 were less than that in P1 group (P＜0.05).Compared with group N1((147.83±60.88) ng/L) and N2((152.73±87.50) ng/L) ,the expression of RAGE protein was increased in group P1((629.89±110.33) ng/L) and P2((229.89±53.20) ng/L) (P＜0.05).There was no significant difference between N1 and N2 groups(P＞0.05) ,while the expression of RAGE protein in P1 group was lower than that of P2(P＜0.05).In immunohistochemical test, positive expression cells in P1 ((18.49± 1.53) and P2 (25.67±3.08)) were higher than that in group N1(9.33±2.31) and group N2(12.14±2.52) (P＜0.05) ,while there was no significant difference between group N1 and N2(P＞0.05).Conclusions Anesthetic dose of propofol can injure spatial learning and memory ability in aged rats.Hyperlipidemia might act synergistically with propofol.

Objective To evaluate the feasibility and advantages of robot-assisted laparoscopic radical prostatectomy (RALRP) in treating high-risk prostate cancer.Methods From Mar.2010 to Dec.2014,69 men with high-risk prostate cancer (clinical stage ≥ T3a,serum PSA ≥ 20 μg/L or biopsy Gleason score ≥8) underwent surgical treatment at our center.RALRP was performed in 44 cases,with a mean age of (66.7 ± 8.6) yrs (range:50 ～ 82) and a mean baseline PSA of (23.1 ± 11.2) μg/L (range:3.6-48.8).Transperitoneal approach was adopted in all cases.Twenty-five patients were treated with ORP.The mean age was (64.3 ± 5.9) yrs (range:52-75) and baseline PSA was (21.7 ± 10.2) μg/L (range:5.7-41.3).Baseline clinical features,including age,initial PSA,biopsy Gleason score and clinical staging,were comparable between two cohorts.Surgical outcomes after RALRP were analyzed and compared between groups.Results All RALRP procedures were successfully performed with da Vinci robotic system and there was no open conversion.Mean operation duration were similarly (158 ± 47)min for RALRP group and (152± 42)min for ORP group,respectively.The mean estimated blood loss and postoperative length of hospital stay in RALRP group were (328 ± 254) ml and (8.4 ± 3.1) days,both of which were significant lower than those in ORP group,(674 ± 302) ml and (14.4 ± 3.7) days.Two cases of urine leak and two of lymphatic leak occurred after RALRP,and a statistically insignificantly higher complication rate was observed for the ORP group,in which four of urine leak and three of lymphatic leak happened.The positive surgical margin rates of two groups were 20.5％ and 24％,respectively.After the mean follow-up of 25 (RALRP) and 27 months (ORP),continence rate was significantly higher in the RARLP group (93.2％ vs.72.0％).PSA failure was observed in seven RALRP cases (15.9％) and four ORP cases (16.0％).Conclusion Compared with traditional ORP,RALRP shows significant advantages,including less blood loss,fewer complications,shorter length of stay,better postoperative continence and equivalent oncologic outcome when dealing with high-risk prostate cancer cases.Robot-assisted surgery is a safe and efficacious surgical modality in this setting.

Objective To discuss the treatment experience of adrenocorticotropic hormone independent macronodular adrenal hyperplasia (AIMAH).Methods The clinical data of 24 cases of AIMAH from August 1972 to December 2016 were retrospectively analyzed.The cases included 10 males and 14 females with a mean age of 43 (range 26 to 59) years.16 patients presented with typical Cushing syndrome (CS) and 8 patients had weight gain, hypertension or diabetes mellitus without any sign of CS.The circadian rhythm of serum cortisol was abnormal.Low and high dose dexamethasone suppression tests failed to suppress cortisol secretion.The preoperative CT scan showed bilateral enlargement of the adrenal glands with multiple macmnodules.14 patients underwent open surgery, including 5 cases of unilateral adrenalectomy, 6 cases of adrenalectomy combined with contralateral subtotal adrenalectomy and 3 cases of bilateral adrenalectomy simuhaneously.6 patients underwent conventional laparoscopic adrenalectomy,including 2 cases of unilateral adrenalectomy, 2 cases of adrenalectomy combined with contralateral subtotal adrenalectomy and 2 cases of bilateral adrenalectomy.4 patients underwent robot-assisted laparoscopic bilateral adrenalectomy.Results The postoperative pathological examination showed that all patients had bilateral adrenal macronodular or adenomatoid hyperplasia.During the mean follow-up of 68 months (range 3 to 192 months), the clinical symptoms of CS disappeared after surgery in all cases.7 patients who received unilateral adrenalectomy had urinary free cortisol and serum cortisol within normal ranges and no further enlargement of the contralateral gland was noticed.Among the 3 patients who received bilateral adrenalectomy simultaneously, 1 case died of adrenal crisis on the seventh day post-operation.The remaining 2 cases presented with adrenal insufficiency but returned to normal after glucocorticoid replacement therapy.Nelson's syndrome was not observed in other patients.Conclusions The use of bilateral adrenalectomy to treat AIMAH may involve risk.Unilateral adrenalectomy is recommended for an alternative treatment for AIMAH.Controlateral adrenalectomy with lifelong corticosteroid replacement or contralateral subtotal adrenalectomy may be performed if the symptoms have not improved or recurred after unilateral adrenalectomy.

Objective To explore the effect of Activating Blood to Resolve Stagnation on the expression of CD34 and vascular endothelial growth factor (VEGF) in rats with acute myocardial infarction. Methods 32 Sprague-Dawley rats were randomly divided into sham operation group (A, n=8), model group (B, n=8), Xuesaitong Injection + granulocyte colony- stimulating factor (G- CSF) group (C, n=8) and G-CSF group (D, n=8). Corresponding medicine was given to each group 3 hours after modeling, for 6 days. Pathomorphological changes were observed through HE staining, and the expression of CD34, VEGF and Ki-67 were observed through immunohistochemical staining. Results The expressions of CD34, VEGF and Ki-67 were higher in groups B, C and D than in group A (P<0.05), and were higher in group groups C and D than in group B (P<0.05). The expressions of CD34 and VEGF were higher in group C than in group D (P<0.05). However, there was no significant difference in the expression of Ki-67 between 2 groups (P>0.05). Conclusion The expression of CD34 and VEGF increases with Activating Blood to Resolve Stagnation method, which is superior to using G-CSF only. Activating Blood to Resolve Stagnation may play an important role in the treatment of acute myocardial infarction.

Objective To summarize our experience in robotic-assisted laparoscopic surgery for adrenal diseases Methods The clinical data of 243 patients with adrenal tumor treated by robotic-assisted laparoscopic surgery from March 2010 to February 2017 were retrospectively reviewed.There were 99 men and 144 women.The mean age was 51.6 years (range, 19-84).Tumors located at left adrenal in 140 cases, right in 97 cases,and both sides in 6 cases.The average diameter was 3.32 cm (range, 0.8-12 cm).However, there were 41 cases whose tumor diameter were greater than 5 cm.Results There were 2 cases of conversion during operation, 1 case converted to open surgery and the other to the traditional laparoscope surgery.The mean operative time was 35 min (range, 20-130 min).Estimated blood loss was 80 ml (range,20-1 200 ml).Blood transfusion was needed in 6 cases.The mean postoperative hospital stay was 5d (range, 3-20 d).The pathological diagnosis included 37 cases of pheochromocytoma, 149 cases of cortical adenoma, 3 cases of cortical carcinoma, 5 cases of metastatic tumor, 9 cases of adrenal myelolipoma, 3 cases of adrenal cyst, 2 cases of bronchogenic cyst, 25 cases of adrenal nodular hyperplasia,2 cases of angiomyolipomas, 1 case of mature teratoma, 1 case of diffuse large B-Cell lymphoma, 1 case of angioma, and 4 cases of neurofibromatosis.Conclusions Robotic-assisted laparoscopic adrenalectomy was safe and effective.Robotic-assisted laparoscopic surgery has the advantages over laparoscopic surgery in treatment of complicated adrenal tumor, such as large adrenal tumors, pheochromocytoma, bilateral adrenal diseases, overweight and obese patients with adrenal diseases.

Due to good mechanical properties and biocompatibility, tissue engineering scaffolds have become the vital method for repairing and regenerating articular cartilage defects. With the continuous development of tissue engineering technology, many scaffolds preparation and formation methods have been developed and tested in the past decade, however, the preparation of ideal regenerative scaffolds remain controversial. As load-bearing tissue inside the body joints, the matrix structure and cell composition of articular cartilage are hierarchical, and there are several smooth natural gradients from the cartilage surface to the subchondral bone layer, including cell phenotype and number, specific growth factors, matrix composition, fiber arrangement, mechanical properties, nutrient and oxygen consumption. Therefore, in the design of regenerative scaffolds, it is necessary to achieve these gradients to regenerate articular cartilage in situ. In recent studies, many new biomimetic gradient scaffolds have been used to simulate the natural gradient of articular cartilage. These scaffolds show different mechanical, physicochemical or biological gradients in the structure, and have achieved good repair effects. The related articles on tissue engineering for the treatment of articular cartilage defects were retrieved by searching databases with key wordsarticular cartilage injury, cartilage repair and gradient scaffolds. In this work,the structural, biochemical, biomechanical and nutrient metabolism gradients of natural articular cartilage were studied and summarized firstly. Then, the latest design and construction of articular cartilage gradient scaffolds were classified. Besides that, the material composition (such as hydrogels, nanomaterials, etc.) and the preparation process (such as electrospinning, 3D printing, etc.) of grandient scaffolds were further enhanced. Finally, the prospect and challenge of biomimetic gradient scaffolds in cartilage engineering are discussed, which provides a theoretical basis for the successful application of gradient scaffolds in clinical transformation.