Agitation plays an important role in the hyaluronic acid (HA) fermentation process. However, views about the effect of agitation on HA production remain controversial. We investigated the effect of agitation on cell growth and HA synthesis during HA fermentation process by using Computational Fluid Dynamics (CFD) technology. The results showed that the biomass and HA yield changed a little with the increase of impeller speed, but the HA molecular weight firstly increased and then decreased. The results of phase agitation control strategy demonstrated that the influence of agitation on the HA molecular weight mainly exhibited at the stage of HA synthesis. Moreover, the CFD simulation results indicated that when impeller speed increased, the mixing time reduced while the shear rate increased significantly. The removal of anchor could moderate the contradiction between the mixing time and shear rate, and finally the HA molecular weight increased by 23.9%. The results of this work could provide guidelines for optimizing the HA fermentation, as well as the bioreactor design and scaling up.
Bioreactors ; Fermentation ; Hyaluronic Acid ; biosynthesis ; chemistry ; Hydrodynamics ; Numerical Analysis, Computer-Assisted ; Shear Strength ; Streptococcus equi ; growth & development ; metabolism

OBJECTIVETo investigate the clinical effects of moxibustion (Mox) in treating knee joint osteoarthritis, and to detect the change of hyaluronic acid (HA) level in serum and synovial fluid (SF) for evaluating its significance.

METHODSThirty OA patients were treated with Mox applied on inner and outer hsiyens and Ashi point for 10 min once a day, 5 times a week for 3 months and the therapeutic efficacy was evaluated. Serum and SF levels of HA were measured by radio-immunoassay before and after the 3-month treatment, and compared with those from 30 non-OA persons for normal control.

RESULTSAfter treatment, 19 patients (20 joints) out of the 30 patients were cured, the efficacy of Mox was evaluated as markedly effective in 8 patients on 10 joints, and as effective in 3 patients on 4 joints, the cure rate being 63.3%. Before treatment, HA level in serum (122.87 +/- 34.10 microg/L) was higher and in SF (0.98 +/- 0.17 g/L) was lower in OA patients than those in the normal controls (68.32 +/- 21.48 microg/L and 1.62 +/- 0.30 g/L, P<0.01), whereas after treatment, both the serum and SF levels of HA in patients changed toward normal range (70.29 +/- 27.30 microg/L and 1.58 +/- 0.26 g/L), showing insignificant difference as compared with those in the controls (P<0.01).

CONCLUSIONMox is an effective approach for treatment of OA, and the levels of HA in serum and SF can be taken as the quantitative indicators for evaluating the pathogenetic condition of OA patients.

Female ; Humans ; Hyaluronic Acid ; analysis ; blood ; Male ; Middle Aged ; Moxibustion ; Osteoarthritis, Knee ; blood ; metabolism ; therapy ; Synovial Fluid ; metabolism

PURPOSE: We aimed to evaluate the overall outcomes of endoscopic dextranomer/hyaluronic acid copolymer (Deflux) injection for vesicoureteral reflux (VUR) and analyze the factors predicting success. MATERIALS AND METHODS: A total 99 patients (58 males, 41 females) and 154 refluxing ureter units were treated with a endoscopic Deflux injection for VUR. At 3 months, radioisotope-voiding cystograms were performed to evaluate treatment responsiveness; success was defined as a resolving of VUR to less than grade I. We evaluated various perioperative factors such as gender, operation age, preoperative antibiotics duration, urinary tract infection, relative renal function and cortical defect, preoperative VUR grade, maximal flow rate in uroflowmetry, laterality of reflux, voiding dysfunction, constipation, orifice shape and trabeculation, injection technique, injection volume, number of punctures, and learning curve. RESULTS: The overall success rate was 62.3% (96/154) in refluxing ureter units (50.5% in patients). According to grade of VUR, the success rate was 87.5% (8/9), 82.2% (37/45), 67.8% (38/56), 33.3% (13/39), and 16.7% (1/6) in grade I, II, III, IV, and V, respectively (p=0.001). In multivariate analysis, preoperative VUR grade and mound morphology were identified as predictive factors (p<0.05). No significant surgery-related complications developed. CONCLUSIONS: Endoscopic Deflux injection for VUR was effective for grade I-III VUR, although the cure rate was low for grade IV-V. The factors predicting success were preoperative VUR grade and mound morphology.
Anti-Bacterial Agents ; Constipation ; Dextrans ; Endoscopy ; Humans ; Hyaluronic Acid ; Learning Curve ; Male ; Multivariate Analysis ; Punctures ; Ureter ; Urinary Tract Infections ; Vesico-Ureteral Reflux

Progressive hepatic fibrosis with development of cirrosis is a feature of chronic liver disease. Assessing fibrosis is important for predicting disease progression and patient management. Liver biopsy is the current gold standard for the diagnosis of liver fibrosis. However, liver biopsy is an invasive procedure. Alternative non-invasive methods have been developed. Serum markers are useful in predicting liver cirrhosis, but accuracy of serum markers is not satisfactory in the assessment of fibrosis. Newly developed transient elastography (Fibroscan) is a non-invasive method of measuring liver stiffness. Fibroscan has been reported to be superior in early detection of cirrhosis to serum markers. Factors influencing it's performance are not fully investigated. The evaluation of new tests should be continued to perform.
Biological Markers/blood ; Chronic Disease ; Disease Progression ; Elasticity Imaging Techniques/methods ; Fibrosis ; Humans ; Hyaluronic Acid/analysis ; Liver/*pathology ; Liver Cirrhosis/*diagnosis/etiology

OBJECTIVETo investigate the effects of sodium hyaluronate on the growth and adhesion of colorectal cancer cells.

METHODSHuman colorectal cancer cell lines SW620 and Colo205 were treated with sodium hyaluronate (25 -2,500 microg/ml), and cancer cell proliferation was measured by MTT assay in vitro. Flow-cytometric analysis was applied to detect expression of CD44 on SW620 and Colo205 cells.

RESULTIn vitro sodium hyaluronate enhanced proliferation of Colo205 cells, but it had no appreciable effect on SW620 growth under the same doses, Meantime, CD44 expression on cancer cells decreased compared with controls.

CONCLUSIONIn vitro sodium hyaluronate has different effects on growth of different colorectal cancer cell lines, but can inhibit CD44 expression of colorectal cancer cells and influence their ability of adhesion.

Cell Adhesion ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Colorectal Neoplasms ; pathology ; Humans ; Hyaluronan Receptors ; analysis ; Hyaluronic Acid ; pharmacology

BACKGROUND: Osteoarthritis (OA), the most common arthritis, is one of the most frequently encountered orthopaedic conditions. As a small number of large joints such as knee and hip are affected in OA, OA is an ideal target for local therapy. Although corticosteroid and hyaluronic acid have been traditionally used for joints through intra-articular (IA) injection, IA injection also provides a minimally invasive route to apply cell therapy to treat OA. IA cell therapy has drawn attention because it may provide regeneration of articular cartilage in addition to palliative anti-inflammatory effects. METHODS: Current progress of IA injection therapy and the author's perspective on this issue are described narratively. RESULTS: It is too premature to have any conclusion on the eventual efficacy of IA cell therapy concerning regeneration of articular cartilage based on current data. Prospective radiological and histological data from larger numbers of patients are needed to prove cost effectiveness of IA cell therapy. CONCLUSION: Expanding research in this field will produce further evidences to provide guidance on the eventual effectiveness of IA cell therapy in the future.
Arthritis ; Cartilage, Articular ; Cell- and Tissue-Based Therapy ; Cost-Benefit Analysis ; Hip ; Humans ; Hyaluronic Acid ; Injections, Intra-Articular ; Joints ; Knee ; Osteoarthritis ; Prospective Studies ; Regeneration

OBJECTIVETo study the inhibitory effect of flavonoids from Glycyrrhiza uralensis on thioacetamide-induced chonic hepatic fibrosis in rats and the effect on the protein expressions of transforming growth factor-β1 (TGF-β1) and Caspase-3 in livers.

METHODMale Sprague-Dawley rats were randomly divided into totally seven groups: the normal control group, the model group, LF groups s (400, 200, 100, 50 mg · kg(-1) · d(-1)) and the silymarin positive control group (30 mg · kg(-1) · d(-1)). The hepatic fibrosis model was induced in the rats through intraperitoneal injection with 3% thioacetamide (TAA) at a dose of 150 mg · kg(-1) body weight twice a week for 12 weeks. During the course, the control group and the model group were orally administered with saline (1 mL · kg(-1) · d(-1)). After the modeling and drug intervention, the pathologic changes and fibrosis in liver tissues were observed by HE staining and Masson's Trichrome staining. The serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and liver hydroxyproline (HYP) contents were assayed by biochemical process. The serum hyaluronic acid (HA) was assessed by radioimmunoassay. In addition, the protein expressions of liver TGF-β1 and Caspase-3 were examined by immunohistochemical method. The mRNA expression of TGF-β1 in hepatic tissues was examined by quantitative Real-time PCR analysis.

RESULTCompared with the model group, flavonoids can protect the integrity of the structure of liver tissues, significantly reduce the hepatic cell degeneration and necrosis and the proliferation of fibrous tissues, notably reduce the serum AST, ALT, ALP and HA and HYP in hepatic tissues and down-regulate the protein expressions of liver TGF-β1 and Caspase-3 and the mRNA expression of TGF-β1 in hepatic tissues.

CONCLUSIONThe licorice flavonoids can resist the thioacetamide-induced hepatic fibrosis in rats. Its mechanism may be related to the down-regulation of the protein expressions of TGF-β1 and Caspase-3.

Animals ; Caspase 3 ; analysis ; Flavonoids ; pharmacology ; Glycyrrhiza uralensis ; chemistry ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; prevention & control ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Thioacetamide ; Transforming Growth Factor beta1 ; analysis ; genetics

BACKGROUNDWe compared the validity (evaluated by sensitivity and specificity), reliability (evaluated by reproducibility) and yield (evaluated by predictive value, examining complexity and cost) of individual and combined tests for bladder tumour antigen stat (BTAstat), nuclear matrix protein 22 (NMP22), hyaluronic acid (HA), survivin, CD44v6, vascular endothelial growth factor (VEGF), and voided urine cytology (VUC) in detecting bladder cancer. And at the same time we evaluated the clinical value of these seven detecting methods in the diagnosis of bladder cancer.

METHODSThe six markers and VUC were detected in the urine of cancer group (151 patients with bladder cancer) and two control groups (50 patients with benign urological diseases and 50 healthy controls). The sensitivity, specificity, predictive value, reproducibility, examining complexity and checking cost of each marker and combined markers were calculated.

RESULTSThere was a significant difference between bladder cancer group and the two control groups. The sensitivity, specificity and positive predictive value were as follows: VUC (36.4%, 100.0%, 100%), BTAstat (76.8%, 87.0%, 89.9%), NMP22 (77.5%, 81.0%, 86.0%), HA (82.8%, 83.0%, 88.0%), survivin (70.2%, 85.0%, 87.6%), CD44v6 (50.3%, 79.0%, 78.4%), and VEGF (68.2%, 93.0%, 93.6%). The highest sensitivities were 91.4% for NMP22 + BTAstat and HA + NMP22, whereas the combined marker with the lowest sensitivity (62.3%) was VUC + CD44v6. The highest specificity was 93.0% for the combined use of VUC + VEGF and HA + CD44v6 had the lowest specificity (73.0%). The most convenient examining method was the detection for BTAstat, the lowest cost was the detection for HA, and the best reproducibility were the detection for BTAstat and VUC.

CONCLUSIONSAll the markers have obvious clinical value in diagnosis of bladder cancer. The use of BTAstat + HA or NMP22 + BTAstat are better examining methods in terms of validity, reliability, and yield.

Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm ; analysis ; Biomarkers, Tumor ; analysis ; Female ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Hyaluronic Acid ; analysis ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; analysis ; Neoplasm Staging ; Nuclear Proteins ; analysis ; Sensitivity and Specificity ; Urinary Bladder Neoplasms ; diagnosis ; pathology ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo observe the influence of intra-articular injection of sodium hyaluronate (SH) on the expression of inducible nitric oxide synthase (iNOS) in the synovium and nitric oxide (NO) content in synovial fluid of rabbits with traumatic osteoarthritis (OA).

METHODSSixteen white rabbits underwent unilateral anterior cruciate ligament transection and were randomly divided into 2 groups 5 weeks after the operation. Rabbits in the experimental group received intra-articular injection of 0.3 ml of 1% SH, once a week for 5 weeks. Animals in the control group were treated under the same conditions using physiological saline. All the animals were sacrificed at the 10th week after surgery. The mRNA expression of iNOS in the synovium was analyzed using reverse transcription-polymerase chain reaction. The content of NO in the synovial fluid was assayed.

RESULTSThe level of iNOS expression of the synovium in the experimental group was lower than that in control group (0.47+/-0.09 vs. 0.65+/-0.12, t equal to 3.45, P less than 0.01). Compared with control group, the content of NO decreased significantly in synovial fluid of SH injection group (134.11 micromolar/L +/- 12.47 micromolar/L vs. 152.17 micromolar/L +/- 15.69 micromolar/L, t equal to 2.55, P less than 0.05).

CONCLUSIONSSH significantly decreases the content of NO in the synovial fluid of rabbits with traumatic OA. SH may exert the effect on synovial fluid NO level as a result of the suppression of iNOS expression in the synovium. It may be one of the mechanisms of the therapeutic effect of SH on early traumatic OA.

Animals ; Anterior Cruciate Ligament Injuries ; Hyaluronic Acid ; pharmacology ; therapeutic use ; Nitric Oxide ; analysis ; Nitric Oxide Synthase Type II ; analysis ; Osteoarthritis ; drug therapy ; etiology ; metabolism ; Rabbits ; Random Allocation ; Synovial Fluid ; chemistry ; Synovial Membrane ; enzymology

Objective: To elucidate the possible role of human lysozyme-like protein 4 (LYZL4) in fertilization and characterize its enzymatic properties. METHODS: The localization of LYZL4 in human spermatozoa was investigated by immunofluorescence staining, the sources of LYZL4 on the sperm surface examined by RT-PCR, and the role of LYZL4 in fertilization assessed by the zona-free hamster egg penetration test. The recombinant plasmid pPIC9K-LYZL4 was constructed and its expression induced with methanol after transformed into competent Pichia pastoris GS115. The recombinant LYZL4 protein (rLYZL4) was purified from the fermentation supernatant and subsequently identified by Western blot. The hyaluronan binding ability of rLYZL4 was determined by ELISA and the muramidase activity, hyaluronidase activity, and free radical scavenging ability examined by spectrophotometric methods. RESULTS: Immunodetection with a specific antiserum localized LYZL4 on the acrosomal membrane of mature spermatozoa, which was exclusively secreted from the testis and epididymis as shown by RT-PCR. Immunoneutralization of LYZL4 significantly decreased the number of human spermatozoa bound to zona-free hamster eggs in a dose-dependent manner in vitro. The recombinant protein was expressed successfully by the P. pastoris strain GS115. Purified rLYZL4 exhibited a potent hyaluronan binding ability and a strong free radical scavenging ability but no muramidase or hyaluronidase activity. CONCLUSIONS LYZL4 secreted from the testis and epididymis is localized on the acrosomal membrane of mature spermatozoa and plays a role in sperm-egg binding as well as in binding hyaluronan and scavenging free radicals, which suggests that it might be a multi-functional molecule contributive to sperm protection and sperm-egg binding.
Acrosome ; enzymology ; Animals ; Blotting, Western ; Cricetinae ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; Female ; Fertilization ; physiology ; Free Radical Scavengers ; metabolism ; Humans ; Hyaluronic Acid ; metabolism ; Male ; Muramidase ; analysis ; physiology ; Pichia ; Plasmids ; metabolism ; Recombinant Proteins ; analysis ; metabolism ; Sperm-Ovum Interactions ; physiology ; Spermatozoa ; enzymology ; Testis