The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

Objective To evaluate the short-term effect of drug-coated balloon (DCB) with paclitaxel in treating in-stent restenosis (ISR) of femoral popliteal artery.Methods A total of 18 patients with femoral popliteal artery ISR,who were admitted to authors' hospital during the period from June 2016 to December 2016 to receive paclitaxel DCB treatment,were enrolled in this study.The patients included 15 males and 3 females,with a mean age of (72.7±9.8) years old.According to Fontaine classification,grade Ⅱ B was seen in 9 patients,grade Ⅲ in 8 patients,and grade Ⅳ in one patient.Preoperative CT angiography showed that ISR could be confirmed in 3 patients and complete occlusion of the stent in 15 patients.After the diseased artery was re-opened,pre-dilatation with common balloon was conducted,which was followed by dilatation with paclitaxel DCB;remedial stent implantation was carried out when the restrictive dissection was observed or if the residual stenosis was beyond 50％.Angiography was performed immediately after the procedure to check the vascular patency.On the third day afterthe treatment,the ankle-brachial index (ABI) of diseased lower extremity was determined to assess the improvement of ischemia.Three month after the treatment,the patients were followed up to detemine the incidence of restenosis,the clinically-driven target lesion revascularization (TLR) rate,and the occurrence of serious clinical events within 3 months.Results Successful re-opening of the diseased artery was achieved in all 18 patients,the technical success rate was 100％.At 3 months after the treatment,the restenosis rate and TLR rate were 10％ (1/10) and 6.67％ (1/15) respectively.No procedure-related complications occurred in all patients.Conclusion For the treatment of femoral popliteal artery ISR,the use of paclitaxel DCB is safe and effective.

Objective To evaluate the effect of TGF-β3 on rabbit nucleus pulposus( NP) cells cultured in three-dimensional polylactic-co-glycolic acid (PLGA)scaffold in vitro.Methods PLGA scaffolds were fabricated by particulate leaching method and soaked in rabbit NP cells suspension(1 × 106/scaffold).PLGA-seeded NP cells were devided into 4 groups: 100 ng/mL TGF-β3/PLGA,500 ng/mL TGF-β3/PLGA,1 μg/mL TGF-β3/PLGA, PLGA control group.Cell proliferation activity was measured using MTT assay.The glycosaminoglycan ( GAG ) analysis were performed by 1, 9-dimethylmethylene blue(DMMB) assay.mRNA expression was measured by quantitive PCR at each time point.Histological observation was performed to elucidate the morphological changes of NP cells in PLGA effected by TGF-β3.Results Higher cellular proliferation activity, GAG production,Collagen type II, Aggrean expression were observed in TGF-β3 /PLGA-seeded NP cells compared with PLGA control group on day-7,day-14,day-21(P<0.05). Higher dose of TGF-β3 exhibited intense cellular proliferation activity and peri-cellular matrix by increasing trend(P<0.05).Histological observation showed TGF-β3/PLGA developed more significant disc cells cluster than PLGA groups on day-21.Conclusion The 3D porous PLGA scaffold-seeded cells using TGF-β3 can promotes cell proliferation, and prompt extracellular matrix(ECM) production.It is a potential biotherapy for the treatment of disc degeneration.

Objective To investigate the effect of intermittent hypoxia (IH) with pulmonary emphysema on the ex-pression of hypoxia inducible factor-1α(HIF-1α),Bax and Bcl-2, and the mechanism underlying the role of HIF-1αin he-patocyte apoptosis thereof. Methods Sixty rats were randomly divided into four groups: normal control group, rats were treated normally;IH group, rats were treated by 30 s nitrogen and then 90 s air, and rats were treated by from 9:00-17:00 daily;pulmonary emphysema group, rats were treated by smudging for half an hour, twice a day (8:00 and 18:00);IH with pul-monary emphysema group, rats were treated by 30 s nitrogen and then 90 s air from 9:00-17:00 daily. After exposure four-teen weeks, rats were killed. qRT-PCR assay was conducted to detect the expression of HIF-1α mRNA, Bax mRNA and Bcl-2 mRNA in live tissues. Results The expressions of HIF-1αmRNA, Bax mRNA and Bax/Bcl-2 were significantly higher in IH with pulmonary emphysema group than those in control group,IH group and pulmonary emphysema group (P<0.05). The expression level of Bcl-2 mRNA was significantly lower in IH with pulmonary emphysema group than that of con-trol group and pulmonary emphysema group (P < 0.05), but no significant difference compared with that of IH group (P >0.05). The levels of HIF-1αand Bax were positively correlated with the level of Bax/Bcl-2 (r=0.732 and 0.699),but the lev-els of HIF-1αand Bax were negatively correlated with the level of Bcl-2 (r=-0.705). Conclusion The expression levels of HIF-1αmRNA, Bax mRNA and Bcl-2 mRNA were over-regulated in hepatocytes induced by intermittent hypoxia with pul-monary emphysema. The HIF-1αexpression was correlated with Bax and Bcl-2, suggesting that HIF-1αmay promote the hepatocyte apoptosis through transcriptional co-activators, Bax and Bcl-2.

Objective To observe the effect of exposure of emphysema with intermittent hypoxia on oxidative stress injury of myocardial cells in rats. Methods Sixty male Wistar rats were divided randomly into four experimental groups(each n=15). The normal control group was bred normally. The emphysema group was exposed to cigarette smoke twice a day(once 30 minutes). The intermittent hypoxia(IH)group was exposed to intermittent hypoxia circumstance 8 hours/day,and the emphysema with IH group was exposed to cigarette smoke twice a day (once 30 minutes)and intermittent hypoxia circumstance 8 hours/day. Each group was exposed for 8 weeks. At the beginning of 9 weeks,the blood gas analysis was performed in 5 rats selected randomly from each group,and the rest rats were sacrificed and their hearts and lungs were taken. Under light microscope,the lung tissues stained with hematoxylin-eosin(HE)were examined. The lung pathology and the results of blood gas analysis showed that the emphysema with IH rat model was established successfully. The levels of malonaldehyde(MDA)and superoxide dismutase(SOD)in rat myocardium were measured by enzyme-linked immunosorbent assay(ELISA),and the subunit p22phox mRNA expressions of nicotinamide adenine dinucleotide phosphate(NADPH)-oxidase were detected by real-time reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with the normal group, the MDA levels and p22phox mRNA expressions were increased obviously in emphysema group, IH group and emphysema with IH group〔MDA(μmol/g):2.93±0.54, 3.58±0.63, 4.51±0.72 vs. 1.75±0.56, p22phox mRNA:0.043±0.004,0.067±0.015,0.123±0.016 vs. 0.018±0.002,all P<0.05〕,but the activities of SOD were decreased significantly(U/mg:36.07±4.79,33.51±7.12,24.29±5.36 vs. 46.08±5.12,all P<0.05). In emphysema with IH group,the increase of MDA levels and p22phox mRNA expressions and decrease of SOD levels were more remarkable compared with those in emphysema group and IH group(all P<0.05). The expression of p22phox mRNA was positively correlated with MDA level(r=0.734,P<0.001). Conclusion The myocardial tissue oxidative stress injury in rats induced by emphysema with intermittent hypoxia exposure is more serious than that induced by exposure of either emphysema or intermittent hypoxia alone,NADPH oxidase possibly being the important medium of myocardial cell response to oxidative stress.

BACKGROUNDThe aim of this research was to evaluate long-term pulmonary sequelae on paired inspiration-expiration thin-section computed tomography (CT) scans 3 years after influenza A (H1N1) virus-associated pneumonia, and to analyze the affecting factors on pulmonary fibrosis.

METHODSTwenty-four patients hospitalized with H1N1 virus-associated pneumonia at our hospital between September 2009 and January 2010 were included. The patients underwent thin-section CT 3 years after recovery. Abnormal pulmonary lesion patterns (ground-glass opacity, consolidation, parenchymal bands, air trapping, and reticulation) and evidence of fibrosis (architectural distortion, traction bronchiectasis, or honeycombing) were evaluated on follow-up thin-section CT. Patients were assigned to Group 1 (with CT evidence of fibrosis) and Group 2 (without CT evidence of fibrosis). Demographics, rate of mechanical ventilation therapy, rate of intensive care unit admission, cumulative prednisolone-equivalent dose, laboratory tests results (maximum levels of alanine aminotransferase, aspartate transaminase [AST], lactate dehydrogenase [LDH], and creatine kinase [CK]), and peak radiographic opacification of 24 patients during the course of their illness in the hospital were compared between two groups.

RESULTSParenchymal abnormality was present in 17 of 24 (70.8%) patients and fibrosis occurred in 10 of 24 (41.7%) patients. Patients in Group 1 (10/24; 41.7%) had a higher rate of mechanical ventilation therapy (Z = -2.340, P = 0.019), higher number of doses of cumulative prednisolone-equivalent (Z = -2.579, P = 0.010), higher maximum level of laboratory tests results (AST [Z = -2.140, P = 0.032], LDH [Z = -3.227, P = 0.001], and CK [Z = -3.345, P = 0.019]), and higher peak opacification on chest radiographs (Z = -2.743, P = 0.006) than patients in group 2 (14/24; 58.3%).

CONCLUSIONSH1N1 virus-associated pneumonia frequently is followed by long-term pulmonary sequelae, including fibrotic changes, in lung parenchyma. Patients who need more steroid therapy, need more mechanical ventilation therapy, had higher laboratory tests results (maximum levels of AST, LDH, and CK), and had higher peak opacification on chest radiographs during treatment are more likely to develop lung fibrosis.

Adult ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; pathogenicity ; Influenza, Human ; complications ; virology ; Lung ; diagnostic imaging ; pathology ; virology ; Male ; Middle Aged ; Pneumonia ; complications ; diagnostic imaging ; microbiology ; Tomography, X-Ray Computed ; methods

Animals ; Athletic Performance ; Drugs, Chinese Herbal ; pharmacology ; Free Radicals ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Morinda ; Myocardium ; metabolism ; Physical Conditioning, Animal

To explore the prevention effect of the joint combination of Yindanxinnaotong soft capsule (YDXNT) and exercise (swimming) on atherosclerotic rats. The method of 3 x 3 factorial design, including two factors (YDXNT and swimming) and three levels (0, 1, 2 g x kg(-1) YDXNT; 0, 0.5, 1 h swimming), was mainly adopted. The atherosclerotic rat model was established by ligating their left common carotid arteries and feeding high-fat diet. After 8 weeks, blood samples were collected from their thoracic aorta to determine blood viscosity, plasma viscosity, fibrinogen (FIB), nitric oxide (NO), 6-keto-PGF(1alpha) endothelin (ET) and thromboxane B2 (TXB2). The tissues of left common carotid arteries of the rats were collected to detect the positive expression of SM22alpha and determine the semi-quantitation through the immunohistochemical staining. The result showed that the combination of YDXNT and swimming can significantly decrease the plasma viscosity (F = 3.241, P = 0.017), the high and low shear blood viscosity (F = 6.444, P = 0.001; F = 3.002, P = 0.024) and FIB (F = 4.046, P = 0.005). The increased NO and 6-keto-PGF(1alpha) and the decreased ET and TXB2 indicated a significant interaction (P < 0.05). The swimming showed an obvious main effect in the expression of up-regulated protein SM22alpha (F = 8.088, P = 0.001). The study suggested that the combined administration of YDXNT and swimming could improve the hemorheological parameters of atherosclerotic rats, protect the vascular endothelium, inhibit the vascular remodeling in atherosclerosis and positively prevent the atherosclerosis.
Animals ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; prevention & control ; Blood Viscosity ; drug effects ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Exercise Therapy ; Humans ; Male ; Microfilament Proteins ; genetics ; metabolism ; Muscle Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Swimming

Objective To evaluate the clinical outcomes of minimally invasive internal fixation for femoral intertrochanteric fracture in osteoporotic elderly patients. Methods A retrospective study was conducted in 112 patients using interan nail (IN) or trochanteric antegrade nail (TAN) for the management of intertrochanteric femoral fracture from January 2009 to September 2015 in our hospital. According to AO classification, there were 34 cases of type 31-A.1, and 61 cases of type 31-A.2, 17 cases of type 31-A.3. Clinical and radiological follow-up were available. Surgical and fluoroscopic time, length of hospital stay, blood loss, complications and hip functions were compared between two groups. Results A total of 78 patients meeting the criteria were evaluated at a mean follow-up of 15 months (range, 3-27 months). The IN was used in 41 patients and the TAN in 37 patients. Operative time, fluoroscopy time and blood loss showed significant difference between the IN group and TAN group (respectively, 58.9±6.9 vs. 75.6±5.9min; 2.70±0.47 vs. 4.40±0.47min; 107.6±6.7 vs. 127.8±6.8ml, P<0.05), suggesting that patients treated with the IN experienced shorter operative and fluoroscopy times, less blood loss and better hip function (73%) than those with TAN (65%, P<0.05). Rate of postoperative complications was lower in the IN group (4.9%) than in the TAN group (10.8%, P<0.05). There was no difference in hospital stay and fracture healing time between the two groups (P>0.05). Conclusions For minimally invasive treatment of unstable femoral intertrochanteric fractures, use of either the IN or TAN is clinically effective. However, IN presents more advantages (e.g., easy operation, reliable fixation, less bleeding, better clinical outcomes, and less complications). The use of IN is a suitable option for the treatment of unstable intertrochanteric fractures in osteoporotic elderly patients.

Objective: To observe the changes in numbers and degranulation status of local mast cells (MCs) of the complementary acupionts, as well as the concentrations of serum histamine and bradykinin after acupuncture at rat's complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 10), for exploring the mechanisms of the synergistic effect of complementary acupionts. Methods: Using random number table method, 40 Wistar rats were randomly divided into a blank control group (group K), an acupuncture at Yanglingquan (GB 34) group (group A), an acupuncture at Yinlingquan (SP 9) group (group B), an acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group AB), and an acupuncture at points near Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group CD), 8 rats in each group. Group A received acupuncture at Yanglingquan (GB 34); group B received acupuncture at Yinlingquan (SP 9); group AB received acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9); group CD received acupuncture at the control points [points 3 mm away from Yanglingquan (GB 34) and Yinlingquan (SP 9) respectively]. Acupuncture was performed at bilateral points in each group. Rats in group K were fixed using the same method as rats in the other 4 groups without acupuncture stimulation. Needles of 0.35 mm in diameter and 40 mm in length were used for acupuncture. Needle handle was connected to the G6805-Ⅱ mode electroacupuncture (EA) device after needling qi was obtained, with sparse-dense wave, frequency of 2 Hz/100 Hz and current of 1 mA to keep the needle handles slightly tremulous while the rats kept quiet. Rats were continuously stimulated by EA for 20 min each time. Experimental interventions were conducted on the 1st, 3rd, 5th and 7th days when the experiment started, for 4 times in total. Specimens from rats in each group were collected 2 h after the 7th day. The levels of histamine and bradykinin in serum were detected by enzyme-linked immunosorbent assay (ELISA). Local tissues of the points were used to prepare cryosections. The changes of MCs were observed after toluidine blue staining. Results: Compared with group K, the numbers and degranulation rates of MCs in group A, group B, group AB and group CD were significantly increased (all P<0.05). The number and degranulation rate of MCs in group AB were significantly higher than those in group A, group B and group CD (all P<0.05). The order of histamine levels from high to low was: group AB >group B > group A > group CD > group K, and the differences were statistically significant (all P<0.05); the levels of bradykinin in group AB, group A and group B were significantly higher than those in group K and group CD (all P<0.05). There was no significant difference in the bradykinin level between group A and group B, nor between group CD and group K (all P>0.05). Conclusion: The number and degranulation rate of MCs of the complementary acupionts are significantly increased after acupuncture at complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 9) of rats, and the serum levels of histamine and bradykinin are increased, which may be one of the mechanisms of the synergism produced by the combination of complementary acupionts.