0.999 5), RSD were no more than 3.0%. The recovery rates were 98.0%-102.0%. There was no significant difference between the results of this method and EDTA-Na2 titration. Conclusion The method is rapid, accurate, simple, little disturbance and is acceptable in determination of total hardness in drinking water.

To improve 3-ketosteroid-△1-dehydrogenase (KSDH) activity and the transformation level for androst-4-ene-3,17-dione,3-ketosteroid-△1 -dehydrogenase gene (ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110 ± 0.5mU and 15 ± 0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B.subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

Human papillomavirus continuous (HPV) infection is an essential factor to induce cervical intraepithelial neoplasia and cervical cancer.Treating HPV infection is considered as a starting point to develop an effective therapeutic vaccine, which is a new strategy for prevention and treatment of cervical cancer.In recent years, development and trials of therapeutic HPV vaccine have made great progress.Selection of vectors, utilization of adjuvants, synthesis of fusion proteins and chimeric proteins have been widely applied to research to enhance vaccine immunogenicity, to increase vaccination safety, to reduce the side effect and so on.The clinical trial results are encouraging: various types of vaccines can induce a specific immune response with good tolerance.However, numerous studies are still required to obtain further success.In addition, HPV exists in various forms, thus it is also the focus of study to expand the range of action and reduce immune escape.

Objective To determine the persistence time of genotype 4 hepatitis E (HE) viremia after the onset of clinical symptoms in HE patients and provide essential data for study on HE epidemiologieal transmission, so that to evaluate potential contagiousness of HE patients after clinical stage. Methods The first serum samples from 162 HE patients after hospitalized in Eastern China were collected and tested for hepatitis E virus (HEV) RNA by nested reversed transcription- polymerase chain reaction (RT-PCR). The persistence time of HEV viremia after the onset of clinical symptoms was estimated with Kaplan-Meier survival analysis. Results HEV RNA was detectable in 101 out of 162 serum samples with positive rate of 62.35%, which was all grouped to genotype 4 by homology analysis. Furthermore, HEV RNA was detectable in 74 (64.91%) out of 114 male and 27 (56.25%) out of 48 female, which was not significantly different (χ2 = 1.08, P=0. 30). Kaplan-Meier survival analysis showed that the median persistence time of HEV genotype 4 viremia was 24 days after the onset of clinical symptoms (95% CI: 18-30 days), which meant that the viremia of 50% HE patients remaining detectable up to 24 days after the onset. The 75% and 25% percentiles were 14 days and 31 days, respectively. There was no significant difference of viremia persistence time between male and female (Breslow test: P=0.98, Tarone-Ware test: P=0.91). Conclusions The viremia of 75% patients with HEV genotype 4 infection could persistent until 2 weeks after the onset of clinical symptoms and that of some patients could persistent over 1 month. It is indicated that the viremia is still persistent and HE patient could be a reservoir even after the clinical symptoms disappeared and biochemical marks normalized.

BACKGROUND: Hydroxyapatite/chitosan (HA/CS) complex may act as a drug carrier for drug release, but little is reported about the release amount and antibacterial effect of minocycline-HA/CS (Mino-HA/CS) complex. OBJECTIVE: To investigate the in vitro release and antibacterial property of Mino-HA/CS complex. METHODS: HA/CS and Mino-HA/CS were prepared using co-precipitation method. The surface and cross-section features of the complexes were observed under scanning electron microscopy. The porosities were measured according to Archimedes Principle. The release of minocycline hydrochloride was measured by high performance liquid chromatography with the simulated saliva as drug release media. In vitro antibacterial effect on Porphyromonas gingivalis and Staphylococcus aureus were measured by bacteria-inhibiting ring method. Biological toxicities were evaluated via cel counting kit-8cel proliferation assay. RESULTS AND CONCLUSION: The porosity of Mino-HA/CS was larger than that of HA/CS, with the average porosity of 53.99%. Single-day release amount of Mino-HA/CS could maintain at the level of 0.5-1 μg per day for a long-term. Bacteriostatic rings of Porphyromonas gingivalis and Staphylococcus aureus stil existed clearly after 7 days. Cel proliferation assays showed that Mino-HA/CS extract had the significant effect on promoting cel proliferation. These findings indicate that the Mino-HA/CS sustains the release of minocycline at a relatively stable level within a longer period, shows good inhibitory effect on Porphyromonas gingivalis and Staphylococcus aureus and promotes the proliferation of periodontal ligament cel s.

Objective To approach the effects of full Marathon on striated muscle and renal function of Marathon amateurs without complaints. Methods A prospective self-paired design study was conducted. The amateurs without body discomfort, hematuria, brown urine, or persistent muscle pain within 1 week after the 2012 Xiamen International Marathon Race were enrolled voluntarily. The peripheral blood and random urine specimens of all subjects under static status 1 week before the race and after the race instantly (within 10 minutes after finishing the race) were collected to detect markers of renal function and striated muscle injury. Results Sixty-one subjects were included in the final analysis of the study with full Marathon of 42.195 km and mean race time of (297.05± 55.60) minutes. Compared with those under static status before the race, the markers of renal function including the levels of urinary N-acetyl-beta-D-glucusamidase [NAG (U/L): 64.00 (54.50, 85.50) vs. 9.50 (8.10, 11.50)], urinary β2-microspheres protein [β2-MG (μg/L): 261.00 (128.50, 1 608.00) vs. 66.60 (33.75, 123.00)], random urinary creatinine [UCr (μmol/L): 19 066.56±10 938.31 vs. 5 872.52±4 363.20] and serum creatinine [SCr (μmol/L): 129.97±25.84 vs. 97.39±14.51] immediately after the race were significantly increased (all P < 0.01); the markers of muscle injury including the levels of serum creatine kinase [CK (U/L): 864.00 (504.00, 1 644.00) vs. 164.00 (128.00, 256.00)], lactic dehydrogenase [LDH (U/L): 383.26±141.69 vs. 182.23±41.12], myoglobin [Mb (mg/L): 1 880.00 (1 080.00, 3 300.00) vs. 42.00 (36.00, 54.50)], alanine aminotransferase [ALT (U/L): 27.0 (19.5, 38.0) vs. 24.0 (15.0, 29.5)] and aspartate transaminase [AST (U/L): 52.07±25.13 vs. 28.28±11.86] were also significantly increased (all P < 0.01), and the increase in CK, Mb, and LDH were more significant. It was shown by correlation analysis that CK after race was negatively correlated with age (r = -0.352, P = 0.005) and body mass index (r = -0.271, P = 0.035), and it was positively correlated with racing time (r = 0.387, P = 0.002) and urinary β2-MG after the race instantly (r = 0.364, P = 0.004). Mb after race was negatively correlated with body mass index (r = -0.331, P = 0.009), and it was positively correlated with urinary β2-MG after the race instantly (r = 0.315, P = 0.013). LDH after race was negatively correlated with age (r = -0.275, P = 0.032) and body mass index (r = -0.377, P = 0.003), and it was positively correlated with urinary β2-MG after the race instantly (r = 0.424, P = 0.001). Conclusion Full Marathon could significantly impact striated muscle and renal function of Marathon amateurs without complaints.

Objective To evaluate the clinical effectiveness of double balloon dilation in percutaneous kyphonplasty on curing vertebral?compression fractures. Methods From January 2009 to September 2013, 84 patients (94 vertebral bodies) with vertebral compression were treated by percutaneous kyphonplasty. All were fresh fractures and were injured or obvious low back pain 1 month, accompanied by local tenderness, kowtow attack painful, lumbar mobility, but no lower extremity injury numbness, activities and defecation disorders. After randomization, the double balloon dilation in percutaneous kyphonplasty method was used to treat 44 patients (49 vertebrae). In the process of performing percutaneous unilateral pedicle puncture and balloon dilata?tion of the vertebral body, the balloon has been moved some distance in the vertebral body. Then completed the perfusion of bone cement, vertebral body forming. 40 cases (45 vertebrae) were used conventional unilateral percutaneous kyphonplasty to vertebtal compression fractures. Recorded the operation time, amount of bleeding, bone?cement injection volume. Used visual analogue scale (VAS), the height of the vertebral body and Cobb angle to evaluated the curative effect. Results All 84 patients completed the operation, follow?up time was 22 months (18-24 months). In two mobile open expansion group, the operation time was about 48 min. The amount of bleeding was 8-15 ml. The average bone?cement injection volume was 5.1 ml. No patients quit the study and no bone cement?leakage cases or other side effects were observed , and no clinical accidents occurred. In a single stretching group, 40 cases (45 vertebrae) completed conventional vertebroplasty, the time of 44 min, bone cement average injection rate 3.2 ml, bleeding 10-15 ml. In two mobile open expansion group, the VAS score was 8.5 points, the height of the vertebral body height was 2.1cm, and the Cobb angle was 34°. After operation, the VAS score 2.9 points, the height of the vertebral body 2.8 cm, and Cobb 20° . In the other group, the pain was significantly relieved and the relief was satisfactor after operation.Vertebral height of 2 cm turned to the last follow?up of 2.4 cm. The Cobb angle was 32°, and the last follow?up was 27°. The VAS score, operation time and bleeding volume of the two groups were not statistically significant, and a statistically significant difference of the average bone ce?ment injection volume, postoperative vertebral height and cobb angle improved with statistical significance. A single open group were 2 cases of bone cement leakage and leakage, 1 cases of bone cement tail, the complication rate was 5.6%. Conclusion The application of double balloon dilation in percutaneous kyphonplasty to vertebral?compression fractures improve relocation of verte?bral compression fractures, increase recovery of vertebral height, and more effectively strengthen and stiffen pathological vertebral bodies, while improving kyphosis. Moreover, it can reduce pressure during bone?cement injections, minimizing the chance of over?flow and leakage, as well as the related side effects, but it will also result in an increase of bone cement?injection volume.

Objective To explore the clinical efficacy and indications of kyphoplasty with movement and secondary en?largement of balloon for the compression fracture of vertebral body with ruptured posterior wall. Methods A retrospective analy?sis was carried out on the data of 29 patients (10 males, 19 females;age range:55-86 years old;mean age:71 years old;29 verte?bral bodies in total) who suffered from compression fracture of the thoracolumbar spine and below, and underwent kyphoplasty through the movement and secondary enlargement of balloon within the vertebral body and were followed up from January 2011 to November 2014. These patients had backache, accompanied by lowered support, limitation of movement, no symptom of nervous lesion on both lower extremities and no past history of balloon kyphoplasty. All fractured vertebral bodies were at T 11 or below, in?cluding 1 case at T11, 4 cases T12, 11 cases L1, 9 cases L2 and 4 cases L3. The causes of injury included fall (19 cases), car accident (8 cases) and unknown reasons (2 cases). All patients underwent kyphoplasty with the movement and secondary enlargement of bal?loon within the vertebral body. Photos were taken immediately after the surgery, at 1 month, 3 months, 6 months and 12 months, and these patients were assessed and analyzed in terms of vertebral height, Cobb angle, visual analogue score (VAS) and Oswestry disability index (ODI). Results The operation time (including the formation and solidification of bone cement) of 29 patients was 40 to 65 min and the mean time was 55 ± 7 min;the blood loss during operation was 2 to 15 ml and the mean blood loss was 5 ± 2 ml;the injected volume of bone cement was 2.5-7.5 ml and the mean volume was 5.5±0.5 ml. Post?operative pain was relieved and ambulation was performed under the protection of lumbar orthosis brace. Statstical analysis was conducted on VAS, ODI, vertebral height and Cobb angle before operation and at 1 month, 3 months, 6 months and 12 months after operation, showing statistically significant differences. X ray examination found that there was no alternation or displacement of bone cement location, and no change in vertebral morphology, the vertebral height and cobb angle remained the post?operative status, and posterior wall rupture of the vertebral body was recovered well. CT revealed that the morphology of bone cement was irregular and closely integrated with bone substance, and no cavity or fissure was seen. Conclusion Kyphoplasty with movement and secondary enlargement of bal?loon within the vertebral body has a good, definite clinical efficacy in treating compression vertebral fracture with incomplete pos?terior wall of the vertebral body without obvious displacement of fractured bone and symptom of nervous lesion on both lower ex?tremities. This surgery is easy to operate, and has an immediate analgesic effect, which could recover vertebral height as well as re?duce kyphosis deformity and improve patient’s prognosis.

Objective:To evaluate the effect of remote ischemic conditioning (RIC) on left ventricular (LV) myocardial perfusion, myocardial viability, LV remodeling, regional and global LV function serially following acute myocardial infarction (AMI) in Chinese mini-pigs.Methods:AMI was established in 12 Chinese mini-pigs (8 males, 4 females; age: 6-8 months) and they were randomly divided into RIC group ( n=6) and non-RIC group ( n=6). RIC was performed in pigs by blood pressure inflation on the lower limbs for 5 min period and 4 cycles immediately after surgery. A series of myocardial perfusion imaging and gated 18F-fluorodeoxyglucose (FDG) myocardial metabolism PET/CT imaging were performed longitudinally at the 1st, 14th, 28th and 56th days after AMI, and parameters including total perfusion defect (TPD), hibernating myocardium (HM), Scar, left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), summed motion score (SMS), summed thickening score (STS) and changes of these parameters were obtained. Independent-samples t test and Mann-Whitney U test were used to analyze data. Results:Nine Chinese mini-pigs survived after surgery and were performed imaging. Compared to non-RIC group ( n=4), HM at the 28th ((6.0±2.4)% vs (17.0±4.6)%; t=-4.158), TPD 14th-1st ((-11.8±5.4)% vs 9.0%(4.5%, 15.0%); z=2.449), TPD 28th-1st ((-15.3±3.9)% vs (12.0±3.0)%; t=-10.071), TPD 56th-1st ((-18.0±6.5)% vs 9.0%(4.5%, 12.0%); z=2.449), HM 28th-1st ((-10.5±6.9)% vs (8.3±2.1)%; t=-4.507), HM 56th-1st (-15.0%(-17.5%, -8.5%) vs 2.0%(0%, 7.0%); z=2.449) and LVEDV 14th-1st (-0.5(-2.5, 0) ml vs (13.0±4.4) ml; z=2.470) were reduced in RIC group ( n=5; all P<0.05). Conclusion:RIC can improve myocardial perfusion, delay LV remodeling in the acute stage and salvage hibernating myocardium in the subacute stage and chronic stage.

As China is actively participating in global health governance in multiple dimensions at multiple levels, Chinese education on global health requires optimizing the curriculum system, strengthening the discipline construction, and improving students' literacy of global health. This study compared two postgraduate courses on global health, Global Health Study: Principles and Practice and Global Health Leadership in Practice, in terms of teaching objectives, course contents, teaching methods, and teaching evaluation. We analyzed the advantages and limitations in the current global heath curriculum, and explored course construction and improvement. We hope to provide a reference for improving the curriculum, teaching, and talent cultivation in the field of global health.