Objective: To clone the Fab gene of a monoclonal antibody (mAb) BDI against human bladder cancer and its expression in E. coli. Methods: Fd and K genes of mAb BDI were cloned by RT-PCR and inserted into an Fab expression vector. Phage displaying Fab and soluble Fab were expressed in E. coli. The N-terminal sequence of VH region was corrected by PCR mediated mutagenesis. The antigen-binding characteristics of the Fab were tested by ELISA and immu-nohistochemistry. Results: Fd and K genes were cloned into the expressing vector p3MH and the phage displaying antibody and soluble Fab were expressed in E. coli, which showed weak binding activity to bladder cancer cells. Correction of the N-terminal sequence of the VHimproved the biding activity dramatically. The feasibility of the application of the Fab in phage antibody library screening was confirmed by a simulated panning procedure. Conclusion: The Fab gene of an anti-human bladder cancer mAb was expressed in E. coli. The importance of the N-terminal sequence on antibody binding activity was suggested.

BACKGROUND: It has been reported that bioactivity is found to be favored by the co-operative behavior of silanol (Si-OH)or Ti-OH etc. groups on the material surface and the involved calcium ions. To confirm the hypothesis that a new family of organic-inorganic hybrid materials, which incorporate gelatin chains covalently into Si-O-Ti network, is synthesized through sol-gel procedure.OBJECTIVE: To synthesize a hybrid of gelatin and CaO-SiO2-TiO2 system which is used for bone repairing, and observe its structure and bioactivity.DESIGN: Randomized control observation.SETTING; the laboratory, School of Material Science and Engineering, Shaanxi University of Science and Technology. MATERIALS: Gelatin (Sinopharm Chemical Reagent Co., Ltd), titanic acid isopropyl ester (Gu'an Hengye Fine Chemicals Co., Ltd), γ-glycidoxy propyl trimethoxy silane (Jingzhou Jianghan Fine Chemicals Co., Ltd), calcium nitrate (Tianjin Bodi Chemicals Co., Ltd)METHODS: The experiment was carried out in the laboratory, School of Materials Sciences and Engineering, Shaanxi University of Science and Technology between April and August in 2006. A new type of bioactive organic-inorganic hybrid was synthesized through sol-gel processing starting from gelatin, γ-(2,3-glycidoxypropyl)trimethoxysilane (GPSM),Tetraisopropyltitanate (TiPT) and calcium nitrate. ①Structure of the hybrid: The structures of the products were investigated with Fourier transformed infrared (FT-IR) diffusive reflection spectroscope, X-ray diffraction (XRD) instrument and scanning electron microscope (SEM) respectively.②Bioactivity: The products Ti15Ca0 and Ti15Ca20 were soaked in a stimulated body fluid (SBF) to evaluate the morphology of the surfaces by SEM and thin-film XRD. MAIN OUTCOME MEASURES: The structure and bioactivity of the hybrid.RESULTS: ①The hybrid was completely amorphous and its surface was almost homogenous, which implied the covalent bonding between the organic component and inorganic component. FT-IR spectra result verified the occurrence of Si-OH group and Si-O-Ti bond, as well as the addition of TiPT supplied Si-O-Ti bond. ②There were lots of apatite crystallines formed on the surface of hybrid Ti15Ca20 after soaked in SBF for 7 days, which confirmed their in vitro bioactivity. These apatite particles were similar to the bioglass and other bioactive materials in the patterns. CONCLUSION: A new type of organic-inorganic hybrid material, which incorporates gelatin chains covalently into Si-O-Ti network, is synthesized through sol-gel procedure. There are lots of apatite and brushite crystals formed on the surface of the product Ti15Ca2O after soaking in SBF for 7 days, which obviously proves the bioactivity of the hybrid.

OBJECTIVE:To observe clinical efficacy and safety of Xuebijing injection combined with imipenem and cilasta-tion in the treatment of severe abdominal infection,and its effects on plasma endotoxin and inflammatory factors. METHODS:Dur-ing Apr. 2013-Apr. 2016,100 patients with severe abdominal infection in our hospital were divided into observation group and control group according to random number table,with 50 cases in each group. Both groups were given Imipenem and cilastation sodium for injection 0.5 g added into 0.9% Sodium chloride injection 500 mL,ivgtt(≥40 min),q12 h. Eight hours later,ob-servation group was additionally given Xuebijing injection 100 mL added into 0.9% Sodium chloride injection 500 mL,ivgtt, bid;Both groups were treated for 5-7 d. The levels of plasma endotoxin and inflammatory factors(TNF-α,IL-6,IL-6/IL-10) were compared in 2 groups before after treatment,and clinical efficacies and the occurrence of ADR was recorded. RESULTS:Before treatment,there was no statistical significance in plasma endotoxin or inflammatory factor levels between 2 groups(P>0.05). After treatment,plasma endotoxin and inflammatory factor levels of 2 groups were decreased significantly,and the obser-vation group was significantly lower than the control group,with statistical significance(P<0.05). The excellent and good rate of observation group was 98.00%,which was significantly higher than 78.00%,with statistical significance(P<0.05). No obvi-ous ADR was found in 2 groups. CONCLUSIONS:Xuebijing injection combined with imipenem and cilastation show significant therapeutic efficacy for severe abdominal infection,can effectively control the release of endotoxin and inflammatory factors with good safety.

BACKGROUND: Fish oil is one of mainly natural resources of n-3 fatty acids, which can inhibit cardiac allograft vasculopathy (CAV) and prolong the survival of cardiac allograft. But, the mechanism is unclear. Recent in vitro data suggested that n-3 fatty acids could inhibit the release of inflammatory transmitter by the activation of peroxisome proliferator-activated receptor-y (PPARy). OBJECTIVE: To test the hypothesis that n-3 fatty acids from fish oil ameliorates CAV development via activating PPARy. METHODS: A total of 6 Lewis rats and 18 Fisher344 rats were randomly selected as heart donors. An additional 24 Lewis rats were randomly and equally divided into 4 groups. In isograft group, heart transplantation was performed among Lewis rats, without any drug. In low-dose fish oil-treated group, F344→Lewis transplantation was performed. At 1 day following surgery, 0.03 mL/kg fish oil was treated by gavage for 8 weeks. In high-dose fish oil-treated group, F344→Lewis transplantation was conducted. At 1 day following surgery, 0.06 mL/kg fish oil was treated by gavage for 8 weeks. In control group, F344→Lawis transplantation was conducted. Cyclosporine A was administrated by gavage for 8 weeks. In the low-dose and high-dose fish oil-treated groups, cyclosporine A (1.5 mg/kg) was given daily by intramuscular injection for 2 weeks following surgery. CAV was evaluated by histological examination. Activity of nuclear factor (NF) k-B and PPARy was assessed in homogenate. Contents of monocyte chemoattractant protein-1 and interferon-inducible protein 10 were measured by enzyme-labeled immunosorbent assay (ELISA). Chemokine receptor CCR2 and CXCR3 expression was determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: All 24 receptor Lewis rats were survived following surgery. The donor heart could regularly beat at 8 weeks following transplantation. Compared with the isograft group, severe CAV was detected in the control group al 8 weeks. Compared with the control group, CAV was significantly relieved, the activity of PPARy was significantly elevated, the activity of NF k-B was significantly decreased, levels of intragraft monocyte chemoattractant protein-1 and interferon-inducible protein-10 were significantly reduced in the low-dose and high-dose fish oil-treated groups (P < 0.001, P < 0.05), especially in the high-dose fish oil-treated group (P < 0.05). There was no significant difference in expression of chemokine receptors CXCR3 in the low-dose and high-dose fish oil-treated groups and control group. Our results demonstrated that n-3 fatty acids from fish oil can attenuate CAV development, possibly through activating PPARy and subsequently inhibiting the NF-kB activation, the chemokines secretion and its receptor expression in a dose-dependent fashion in rat models.

Objective To research the protective effect of Ento-Ⅰagainst cerebral ischemia-reperfusion injury in rats,and to evaluate its analgesic and anticoagulating effects in mice. Methods The ischemic model was established with line embolism to block the middle cerebral artery of male rats. The 56 rats were randomly assigned into 7 groups of sham-operation,blank-matrix,nor?
mal saline,Ento-Ⅰplastic of 3 doses(6.67,3.33,1.67 mg/kg),and ozagrel sodium(8.3 mg/kg,ip). The effect of Ento-Ⅰplastic on anti-cerebral ischemia was measured by nervous function scores and the areas of cerebral infarction were determined by TTC staining for the calculation of cerebral infarction rates. The analgesic effect of Ento-Ⅰplastic was determined with acetic acid-induced twisting experiment. Sixty KM mice were randomly allocated into blank-matrix,aspirin,aspirin-plastic,and Ento-Ⅰplastic of 3 doses(5,10 and 20 mg/kg),the number of mouse twisting were recorded right after intraperitoneal injection of 0.7%acetic acid solution at the time of 1 h after the last administration. Moreover,the anticoagulant activity of Ento-Ⅰplastic was tested by glass capillary method. Re?sults The results of acetic acid-induced twisting experiment displayed that Ento-Ⅰplastic of all 3 dose groups(5,10 and 20 mg/kg) could significantly reduce the number of body torsion and increase the inhibitory rates of twisting,compared with that of blank matrix group(the inhibitory rates of twisting for 3 dose groups were 21.79%,48.89%,and 56.15%,respectively),with dose-response man?ner. According to the results of glass capillary test,the clotting time of mouse blood could be significantly prolonged by mid-(10 mg/kg)and low-dose(5 mg/kg)of Ento-Ⅰplastic with corresponding clotting time of(155.20±54.19)s and(155.80±73.84)s,compared with normal saline group at(92.10 ± 24.61)and blank-matrix group at(80.40 ± 48.09,P<0.05). The experiment results of the isch?emia-reperfusion injury by line embolism method in rats exhibited that Ento-Ⅰplastic in mid-dose(3.33 mg/kg)could significantly re?duce the neurological scores after 24 h of reperfusion injury,from(2.33 ± 0.52)of normal saline group to(1.00 ± 0.00)of mid-dose group(P<0.01). The results from TTC staining revealed that the cerebral infarction rates of normal saline group and blank-matrix group were(24.89±7.24)%and(27.72±7.89)%,respectively,whereas those of 6.67 mg/kg and 3.33 mg/kg group of Ento-Ⅰplastic were(14.01±2.65)%and(14.73±4.94)%,respectively. Compared to the 2 negative-control groups,both the high-and mid-dose of Ento-Ⅰplastic could significantly reduce the cerebral infarction rates after ischemic reperfusion injury in rats (P<0.01). Conclu?sion Ento-Ⅰplastic demonstrates strong analgesic and anticoagulant effects,and could substantially reduce the neurological scores and reduce cerebral infarction rates for ischemia-reperfusion injured rats. These are likely to be the mechanism of action for Ento-Ⅰplastic realizing its anti-cerebral ischemia effect.

Objective To systematically evaluate the diversity of oral flora in patients with pancreatic cancer. Methods A cross-sectional study was conducted, focusing on the oral flora diversity profiles of patients with pancreatic cancer. The studies were retrieved from PubMed, Web of science, EMbase, The Cochrane Library, CBM, CNKI, Wanfang, and VIP databases, and the search period was from the establishment of the database to July 15, 2023. According to the inclusion and exclusion criteria, two researchers screened intensive review literature, extracted data and information, and carried out Meta-analysis using qualitative systematic review and Review Manager 5.4. Results Seven cross-sectional studies were reviewed, including 187 patients with pancreatic cancer and 440 healthy controls. The results of meta-analysis showed that the oral microbiota diversity Simpson index of patients with pancreatic cancer was reduced compared with that of healthy controls. Qualitative analysis showed that the relative abundance of Firmicute, Prevotella, Roseburia, and Streptococcus in patients with pancreatic cancer was higher than that in healthy people. The relative abundance of Proteobacteria, Neisseria, Haemophilus, porphyromonas, and Haemophilus parainfluenza in patients with pancreatic cancer was lower than that in healthy people. Conclusion Patients with pancreatic cancer have distinct oral flora, which has high relative abundance of Firmicutes, Prevotella etc. and low relative abundance of Proteobacteria, Neisseria, etc.

OBJECTIVETo prepare effervescent osmotic pump tablet (EOPTs) according to the rhythm of coronary heart disease based on efficacy material and the mechanism of compound Danshen and to study the mechanism of drug released of that tablets.

METHODSince compound Danshen consist of compounds with polyphenolic groups or carboxyl groups, such as phenolic acids, flavonoids, and triterpenoids that they were acidic. EOPTs were prepared from tablet cores which containing NaHCO3 as effervescent, NaCL and manitol as osmotic agents, HPMC as retarding agents coating with CA membrane. And study the mechanism of drug released according to the change of tablet osmotic pressure.

RESULTThe results of in vitro experiments showed that no difference was observed among the profiles of Danshensu, protocatechuic aldehyde, ginsenoside Rg1, Rb1, notoginsenoside R1 release EOPTs. The drug was completely released from the device with a zero-order release rate over 12 h.

CONCLUSIONEOPTs are Successfully obtained EOPT which the drug is released from the device over 12 h and the release mechanism of EOPTs is explained.

Coronary Disease ; physiopathology ; Drug Compounding ; Drugs, Chinese Herbal ; administration & dosage ; metabolism ; Infusion Pumps ; Osmosis ; Salvia miltiorrhiza ; metabolism ; Tablets ; Time Factors

OBJECTIVETo prepare pulsed-release tablet (PTS) according to the rhythm of coronary heart disease based on efficacy material and the mechanism of compound Danshen.

METHODPTS were achieved by coating the core which contains drugs, CMS-Na, lactose, succinic acid and MCC with separation layer (Eudragit RL), swelling layer (HPMC E5), and controlled-release membrane (Eudragit RS-RL-EC).

RESULTThe results of in vitro experiments showed that no difference was observed among the profiles of Danshensu, protocatechuic aldehyde, ginsenoside Rg1, Rb1, notoginsenoside R1 release from the two-step release system. And it indicated that swelling was the basis and prerequisite for drug release from PTS, and the diffusion, organic acid-induced, and osmotic pumping mechanism were involved in drug release, but the latter they were the dominant factors.

CONCLUSIONSuccessfully obtained the PTS of a certain lag-time behind the rapid release which indicate that after bed time administration of such device, the drug plasma concentration-time curve CAN meet the requirements of chronotherapy of cardiovascular disease.

Benzaldehydes ; metabolism ; Catechols ; metabolism ; Chromatography, High Pressure Liquid ; Coronary Disease ; drug therapy ; Diffusion ; Drug Compounding ; methods ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; metabolism ; therapeutic use ; Ginsenosides ; metabolism ; Osmosis ; Salvia miltiorrhiza ; chemistry ; Tablets ; Time Factors

Objective·To investigate the effect of Astragali Radix on T lymphocyte subsets and cytokine expression in Hashimoto's thyroiditis patients with normal thyroid function.Methods·A total of 120 Hashimoto's thyroiditis patients with normal thyroid function and complete data were selected from January 2020 to December 2020 in Jinshan Branch of Shanghai Sixth People's Hospital.The patients were randomly divided into intervention group(n=60)and control group(n=60)by the method of random number table.The treatment plan of the control group was iodine appropriate state diet,and the intervention group was combined with oral Astragali Radix solution(150 mL per time,twice/d)on the basis of the treatment of the control group.The two groups were treated for 6 months.The changes in peripheral blood serum T lymphocyte subsets(CD3+,CD4+,CD8+,and CD4+/CD8+),cytokines[interleukin-2(IL-2),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-10(IL-10)],hypersensitive C-reactive protein(hs-CRP),erythrocyte sedimentation rate(ESR),thyroid function,autoantibody,liver and kidney function,and other biochemical indexes were compared before and after treatment between the two groups.Adverse reactions were observed during the treatment.The factors influencing the change amplitude of thyroid peroxidase antibody(TPOAb)were analyzed by multifactor linear regression.Results·Finally,118 patients,with 59 cases in each group,were included in the study.After 6 months of treatment,the intervention group showed significant improvements in the proportions of CD4+ T cells,the ratio of CD4+/CD8+,and the levels of IL-2,TNF-α,IL-10,hs-CRP,ESR,TPOAb,and thyroglobulin antibody(TGAb)compared to the values before treatment and in the control group(P<0.05).There were no statistically significant differences on the above indicators before and after treatment in the control group(P>0.05).No serious adverse reactions were observed in the intervention group.Multiple linear regression analysis results showed that the use of Astragali Radix,increase of CD4+ level,increase of CD4+/CD8+ ratio,and decrease of hs-CRP level were influencing factors for the decrease of TPOAb level(β=-0.393,P=0.029;β=-0.513,P=0.010;β=-0.351,P= 0.035;β=0.434,P=0.023).Conclusion·Astragali Radix can improve the levels of CD4+ T cells,CD4+/CD8+ratio,IL-2,TNF-α,IL-6,and IL-10 in Hashimoto's thyroiditis patients with normal thyroid function,and it is safe to use.