AIM To study the synergistic anticancer effccts of neferine combined with anticancer agents on Saos 2 cell lines and its mechanism. METHODS\ Cytotoxicity assay was tested by MTT method.Cell cycle and apoptosis was detected by PI staining flow cytometty. Expression of Rb protein was examined by indirect immune fluorescent symbol flow cytometty and S P immunohistochemical staining. RESULTS\ 5,10 ?mol?L -1 Nef augmented apoptosis of Saos 2 cells induced by ADM,enhanced the expression of Rb protein in Saos 2 cells and raised the G 1 phase ratio of Saos 2 cells. CONCLUSION Nef can promote chemotherapy sensitivity to ADM in Saos 2 cells. Its mechanism is associatd with the up modulation of Rb protein expression in the Saos 2 cells arrest cells in G 1 phase

Sucrose phosphorylase (SPase) gene from Leuconostoc mesenteroides ATCC 12291 was synthesised after codon optimization, and inserted into pET-28a plasmid to generate pET-28a-spase. The recombinant strain Escherichia coli BL21 (DE3)/pET-28a-spase was induced for Spase expression. The recombinant protein Spase was purified and characterized. The specific enzyme activity of SPase was 213.98 U/mg, the purification ratio was 1.47-fold, and the enzyme activity recovery rate was 87.80%. The optimal temperature and the optimal pH of the SPase were identified to be 45 °C and 6.5 respectively, and Km, Vmax and kcat of the SPase for sucrose was 128.8 mmol/L, 2.167 μmol/(mL·min), and 39 237.86 min-1. The recombinant SPase was used for α-arbutin production from hydroquinone and the reaction process was evaluated. The optimal conditions for synthesis of α-arbutin by SPase were 40 g/L hydroquinone, 5:1 molar ratio of sucrose and hydroquinone, and 250 U/mL recombinant SPase at pH 7.0 and 30 °C for 24 h in the dark, and then 500 U/mL glucoamylase was added at 40°C for 2.5 h. Under the optimized process, the yield of α-arbutin reached 98 g/L, and the hydroquinone conversion rate was close to 99%. In summary, the recombinant SPase was cloned and characterized, and its application for α-arbutin production was feasible.

Limonene and its derivative perillic acid are widely used in food, cosmetics, health products, medicine and other industries as important bioactive natural products. However, inefficient plant extraction and high energy-consuming chemical synthesis hamper the industrial production of limonene and perillic acid. In this study, limonene synthase from Mentha spicata was expressed in Saccharomyces cerevisiae by peroxisome compartmentalization, and the yield of limonene was 0.038 mg/L. The genes involved in limonene synthesis, ERG10, ERG13, tHMGR, ERG12, ERG8, IDI1, MVD1, ERG20ww and tLS, were step-wise expressed via modular engineering to study their effects on limonene yield. The yield of limonene increased to 1.14 mg/L by increasing the precursor module. Using the plasmid with high copy number to express the above key genes, the yield of limonene significantly increased up to 86.74 mg/L, which was 4 337 times higher than that of the original strain. Using the limonene-producing strain as the starting strain, the production of perillic acid was successfully achieved by expressing cytochrome P450 enzyme gene from Salvia miltiorrhiza, and the yield reached 4.42 mg/L. The results may facilitate the construction of cell factory with high yield of monoterpene products by S. cerevisiae.
Saccharomyces cerevisiae/metabolism* ; Limonene/metabolism* ; Metabolic Engineering ; Monoterpenes/metabolism*

The limitations of antifungal drugs and severe drug resistance make the treatment of invasive fungal infections (IFIs) a great challenge. Itraconazole (ITZ), as a clinical first-line drug, has a wide range of antifungal activity, but it is still limited by adverse reactions such as liver and kidney toxicity, headache and abdominal pain due to its poor water solubility and easy to cause drug accumulation by injection. In this study, the amphiphilic polymer gallic acid-chitosan-cinnamaldehyde (GA-CS-CN) was prepared by amide reaction and Schiff-base reaction. The drug-loaded nanoparticles (GA-CS-CN/ITZ) were prepared by ultrasonic method. The properties of nanoparticles formulations and its in vitro antifungal activity were investigated in the study. Studies have shown that GA-CS-CN/ITZ is spherical, homogeneous and stable, and has good biological safety. The average particle size was 239.57 ± 31.37 nm, ITZ encapsulation efficiency was (93.41 ± 1.12)%, and the cumulative drug release was (62.25 ± 1.88)% at 48 h in vitro. The antifungal activity results of Candida albicans ATCC 10231 (C. albicans) showed that it had the optimal antifungal effect and could significantly enhance the antifungal activity of free drugs. GA-CS-CN/ITZ prepared in this study has excellent biological safety and anti-C. albicans performance, which provides a new choice for the treatment of C. albicans infection and has good application potential in the treatment of this fungal infection.

OBJECTIVE: To evaluate the clinical outcomes of reverse total shoulder arthroplasty as a revision procedure for the failed fixation of proximal humeral fractures in the elderly patients. METHODS: A retrospective analysis was performed on 8 patients with failed internal fixation of proximal humeral fractures from May 2014 to March 2020, including 3 males and 5 females, aged from 65 to 75 years old. All 8 patients underwent reverse total shoulder arthroplasty, and the mean time between initial fixation and reverse total shoulder arthroplasty ranged from 8 to 16 months. Range of motion(ROM), University of California at Los Angeles(UCLA) shoulder score, visual analogue scale (VAS), self-rating anxiety scale(SAS), and Constant-Murley score of shoulder function were assessed pre-operatively and at the last follow-up. Complications relating to the surgery were recorded. RESULTS: All 8 patients successfully followed up. The mean follow-up after reverse total shoulder arhroplasty ranged from 16 to 28 months. The range of motion (forward flexion, external rotation, abduction and internal rotation) of the affected shoulder was significantly improved after surgery, and the post-operative VAS, SAS and UCLA scores were also significantly improved. For the Constant-Murley score of shoulder joint function, the total scores and the subscores of pain, daily activities, range of motion and strength test at the last follow-up were all significantly improved. Scapular glenoid notch was observed in patient, which was evaluated as grade 1 on imaging. All the other patients did not develop specific or non-specific complications. CONCLUSION Reverse total shoulder arhroplasty is an appropriate treatment as a revision surgery for failed fixation of proximal humeral fractures. It has shown satisfactory clinical outcomes, accelerating the rehabilitation of shoulder function and improving the quality of life.
Male ; Female ; Humans ; Aged ; Shoulder/surgery* ; Arthroplasty, Replacement, Shoulder/methods* ; Retrospective Studies ; Treatment Outcome ; Quality of Life ; Shoulder Joint/surgery* ; Shoulder Fractures/surgery* ; Humerus/surgery* ; Range of Motion, Articular

Arthroplasty, Replacement, Shoulder ; Humans ; Humeral Head ; Shoulder Fractures ; surgery ; Shoulder Joint

OBJECTIVE: To explore the clinical significance of elongated needle in the treatment of urinary retention after spinal cord injury. METHODS: From January 2016 to June 2018, 60 patients with urinary retention after spinal cord injury were divided into elongated needle acupuncture group and needle acupuncture group, 30 cases in each group. The Shuidao(ST28) and Zhibian(BL54) were selected as the treatment of acupoints. In the elongated needle group, there were 23 males and 7 females, with a mean age of(52±9) years old. The patients in this group were treated on both sides, and the electro-acupuncture apparatus was used for 30 minutes at frequencies of 3 Hz for 30 minutes. In needle group, there were 24 males and 6 females, with a mean age of (56±10) years old. After treatment, the needles were retained for 30 minutes. The treatment frequencies of both groups were once every other day and passed for 2 months. After 2 months of treatment, the clinical significances of the treatment of the disease were explored by analyzing the time between the two groups of patients to reach the equilibrium bladder, diary urinary diary, urodynamics and urinary tract infection between the two groups. RESULTS: (1)Time to reach the equilibrium bladder:(39.5±1.2) days in elongated needle group and (46.5±2.1) days in needle group; the time to reach the equilibrium bladder in the two groups was significantly different(<0.05). (2)Comparison of urination diary between the two groups:the daily number of catheterization, the amount of each catheterization, the number of daily urination, and the amount of urine per urination after treatment were better than those before treatment by intra-group comparison(<0.05); the number of daily catheterization, the amount of daily catheterization, the number of daily urination, and the amount of urine output per patient in elongated needle group were better than those in needle group by comparison between two groups(<0.05). (3)Comparison of improvement of urinary tract infection between the two groups:the urinary tract infection were better than that before treatmant in each group by intra-group comparison; the improvement rate of urinary tract infection in awn needle group (1/3.3%) was higher than that in needle group(5/16.7%), with significantly differences. (4)Comparison of urodynamic index between two groups:by intra-group comparison, the VH2O, Pves, Pdet, Qave, Qmax, and BC of the two groups were better than those before treatment(<0.05); By comparison between two groups, the improvement of VH2O, Pves, Pdet, Qave, Qmax and BC were better than those of the needle group (all <0.05). CONCLUSIONS The elongated needle is better than conventional needle in improving urinary retention, urinary tract infection and urodynamic index in patients with urinary retention after spinal cord injury. It has positive significance in improving patients' quality of life and is worth applying to clinical.
Adult ; Female ; Humans ; Male ; Middle Aged ; Needles ; Quality of Life ; Spinal Cord Injuries ; Urinary Retention ; Urodynamics

Objective To investigate celluar toxicity and antibacterial property of nano-silver coating modified denture base and to provide a theortical basis for clinical application.Method The samples were divided into coating group and denture base group.Samples in coating groups were treated with silver ion sputtering and were further divided into the 10,20,40,60,80,100 s subgroups according to their sputtering time.Surface microstructure of samples in coating groups and denture base group were observed by scanning electron microscopy (SEM),and composition of material surface were determined with energy dispersive spectrum (EDS) analysis.Methyl thiazolyl terazolium(MTT) method was used to analysis L929 cells proliferation rate and cell toxicity grade while the coating groups(n =6) and group test specimens (n =6),negative control (polyethylene,PE) (n =6) and positive control group (PVC-org.Sn) (n =6) were co-cultured with L929 cells.The antibacterial properties of coating groups were investigated by using the bacterial membrane adhering method with Streptococcus mutans and Candida albicans.Results SEM observation showed that the surface of control group specimens were smooth,whereas dense and uniform distributed nano silver particles were visible on the surface of coating group specimens.The silver particles were round,and the diameter size was 10 nm.MTT assay showed that after being co-cultured with specimens in control group,L929 cell had higher proliferation rate than 100％ at 24,48,72 h,and cytotoxicity were graded 0.The membrane adhering method results showed that all of bacterial colonies were found in control group,but without anything at the coating groups.Conclusions Deposition of nano silver on the denture base surface by ion sputtering method is a viable method.There's no cytotoxicity of series Ag-coating material.Nano-silver modified denture base material benefits L929 cell surface proliferation and has antibacterial action.

Bacillus subtilis is recognized as a generally-regarded-as-safe strain, and has been widely used in the biosynthesis of high value-added products, including N-acetylneuraminic acid (NeuAc) which is widely used as a nutraceutical and a pharmaceutical intermediate. Biosensors responding to target products are widely used in dynamic regulation and high-throughput screening in metabolic engineering to improve the efficiency of biosynthesis. However, B. subtilis lacks biosensors that can efficiently respond to NeuAc. This study first tested and optimized the transport capacity of NeuAc transporters, and obtained a series of strains with different transport capacities for testing NeuAc-responsive biosensors. Subsequently, the binding site sequence of Bbr_NanR responding to NeuAc was inserted into different sites of the constitutive promoter of B. subtilis, and active hybrid promoters were obtained. Next, by introducing and optimizing the expression of Bbr_NanR in B. subtilis with NeuAc transport capacity, we obtained an NeuAc-responsive biosensor with wide dynamic range and higher activation fold. Among them, P535-N2 can sensitively respond to changes in intracellular NeuAc concentration, with the largest dynamic range (180-20 245) AU/OD. P566-N2 shows a 122-fold of activation, which is 2 times of the reported NeuAc-responsive biosensor in B. subtilis. The NeuAc-responsive biosensor developed in this study can be used to screen enzyme mutants and B. subtilis strains with high NeuAc production efficiency, providing an efficient and sensitive analysis and regulation tool for biosynthesis of NeuAc in B. subtilis.
N-Acetylneuraminic Acid/metabolism* ; Bacillus subtilis/metabolism* ; Promoter Regions, Genetic/genetics* ; Binding Sites ; Biosensing Techniques

OBJECTIVE: To investigate clinical results of reverse total shoulder arthroplasty in treating old fracture of proximal humerus in elderly patients. METHODS: From January 2012 to December 2017, 12 elderly patients with old proximal humeral fractures were treated with reverse total shoulder arthroplasty. There were 5 males and 7 females with an average age of 70.2 years old (ranged from 63 to 81 years old) and an average course of 12.3 months (ranged from 9 to 18 months). VAS score was used to evaluate the degree of pain relief of shoulder joint, Constant-Murley score was used to evaluate the improvement of shoulder joint function and observe the complications during and after operation. RESULTS: All the 12 patients were followed up with an average duration of 22.3 months (ranged from 15 to 56 months). At the latest follow-up, the VAS score was 1.9±1.4, and Constant-Murley score was 83.4±8.4. Among them, shoulder joint flexion was (92.5±7.7)°, abduction was (90.4±14.3)°, external rotation was(31.9±10.0)°, and internal rotation was(58.9±13.1)°. There was statistical significance before and after treatment(<0.05). There were 2 cases with glenoid notch. The imaging findings were grade I and grade II respectively. No loosening was found during follow-up. No infection, acromial fracture, prosthesis loosening, axillary nerve injury and other complications occurred. CONCLUSIONS Reverse total shoulder arthroplasty in treating old proximal humeral fractures in elderly patients can achieve satisfactory results, however, indications and complications should be noted.
Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Shoulder ; Female ; Humans ; Humerus ; Male ; Middle Aged ; Shoulder ; Shoulder Fractures ; surgery ; Shoulder Joint ; Treatment Outcome