Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

Jiuwei Xifeng Granules have become a Chinese patent medicine in the market. Because the formula contains Os Draconis, a top-level protected fossil of ancient organisms, the formula was to be improved by replacing Os Draconis with Ostreae Concha. To evaluate whether the improved formula has the same effectiveness and safety as the original formula, a randomized, double-blind, parallel-controlled, equivalence clinical trial was conducted. This study enrolled 288 tic disorder(TD) of children and assigned them into two groups in 1∶1. The treatment group and control group took the modified formula and original formula, respectively. The treatment lasted for 6 weeks, and follow-up visits were conducted at weeks 2, 4, and 6. The primary efficacy endpoint was the difference in Yale global tic severity scale(YGTSS)-total tic severity(TTS) score from baseline after 6 weeks of treatment. The results showed that after 6 weeks of treatment, the declines in YGTSS-TSS score showed no statistically significant difference between the two groups. The difference in YGTSS-TSS score(treatment group-control group) and the 95%CI of the full analysis set(FAS) were-0.17[-1.42, 1.08] and those of per-protocol set(PPS) were 0.29[-0.97, 1.56], which were within the equivalence boundary [-3, 3]. The equivalence test was therefore concluded. The two groups showed no significant differences in the secondary efficacy endpoints of effective rate for TD, total score and factor scores of YGTSS, clinical global impressions-severity(CGI-S) score, traditional Chinese medicine(TCM) response rate, or symptom disappearance rate, and thus a complete evidence chain with the primary outcome was formed. A total of 6 adverse reactions were reported, including 4(2.82%) cases in the treatment group and 2(1.41%) cases in the control group, which showed no statistically significant difference between the two groups. No serious suspected unexpected adverse reactions were reported, and no laboratory test results indicated serious clinically significant abnormalities. The results support the replacement of Os Draconis by Ostreae Concha in the original formula, and the efficacy and safety of the modified formula are consistent with those of the original formula.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Double-Blind Method ; Drugs, Chinese Herbal/therapeutic use* ; Tic Disorders/drug therapy* ; Treatment Outcome

OBJECTIVE: To explore clinical effect of bone cement-strengthened pedicle screw technique in the correction of stage Ⅲ asymptomatic Kümmell disease with kyphosis. METHODS: A retrospective analysis was conducted on clinical data of 40 asymptomatic stage Ⅲ Kümmell disease patients admitted between March 2019 and December 2021, including 15 males and 25 females, aged from 61 to 81 years old with an average of (67.4±5.2) years old;according to different surgical methods, they were divided into percutaneous kyphoplasty group (PKP) and reinforced screw group. There were 18 patients in PKP group, including 7 males and 11 females, aged from 61 to 78 years old with an average of (66.2±5.5) years old;the courses of disease ranged from 5 to 12 months with an average of (7.33±1.78) months;bone mineral density(BMD) T values ranged from -2.45 to -4.00 with an average of (-3.08±0.46);2 patients with T₈-T₉, 10 patients with T₁₀-T₁₂, and 6 patients with L₁-L₂;treated with PKP. There were 22 patients in reinforced screw group, including 8 males and 14 females, aged from 65 to 81 years old with an average of (68.5±3.8) years old;the courses of disease ranged from 4 to 15 months with an average of (7.86±2.73)months;bone mineral density(BMD) T values ranged from -2.40 to -4.50 with an average of (-3.18±0.54);3 patients with T₈-T₉, 12 patients with T₁₀-T₁₂, and 7 patients with L₁-L₂;treated with bone cement reinforced pedicle screw internal fixation combined with kyphoplasty. Cobb angle and anterior margin height of the injured vertebra were compared before operation, 3 d and 12 months after operation. Visual analogue scale (VAS) and Oswestry disability index (ODI) were compared between two groups before operation and 12 months after operation. The incidence of postoperative complications was compared between two groups. RESULTS: All patients were followed up, PKP group followed up for 11 to 14 months with an average of (11.97±0.96) months and 10 to 14 months with an average of (12.05±1.09) months in reinforced screw group;there was no significant difference between two groups (P>0.05). Postoperative Cobb angle at 3 days and 12 months in reinforced screw group were (7.34±2.26) ° and (18.86±1.96) °, while in PKP group were (18.88±1.89) ° and (23.28±1.90) °;there were statistical difference between two groups (P<0.05). The anterior margin height of the injured vertebra in reinforced screw group were (25.28±1.33) mm and (19.62±2.22) mm at 3 days and 12 months after operation, while in PKP group were (18.61±2.16) mm and(15.93±1.34) mm;there were statistical difference between two groups (P<0.05). Cobb angle and the anterior margin height of the injured vertebra were significantly improved at 3 days and 12 months after operation between two groups (P<0.05). Postoperative VAS and ODI at 12 months in PKP group were (2.00±0.69) score and (13.44±4.02)%, while in reinforced screw group were(1.91±0.61) score and (10.18±4.26)%;which were significantly lower than (6.89±0.76) score and (36.33±3.40)% in PKP group, (7.23±0.75) score and (37.09±3.73) % in reinforced screw group before operation. There were no difference in postoperative VAS between two groups at 12 months (P>0.05);postoperative ODI in reinforced screw group at 12 months was lower than that in PKP group(P<0.05). There was no significant difference in complications between two groups (χ²=0.071, P>0.05). CONCLUSION PKP and bone cement reinforced nail combined with PKP could improve kyphotic deformity and postoperative function, and relieve pain. The application of bone cement-reinforced nail fixation technology could provide a more stable support, more obvious functional recovery, lower the risk of re-collapse of the injured vertebra, and maintain the long-term stability of spine.
Humans ; Male ; Female ; Aged, 80 and over ; Kyphosis/surgery* ; Aged ; Bone Cements ; Middle Aged ; Retrospective Studies ; Pedicle Screws ; Spinal Fractures/surgery* ; Fracture Fixation, Internal/methods* ; Bone Screws ; Kyphoplasty

AIM To establish a quantitative analysis of multi-components by single-marker(QAMS)method for the simultaneous content determination of gastrodin,parishin E,syringin,parishin B,parishin C,ferulic acid,parishin A,buddleoside,harpagoside and cinnamic acid in Tianma Toufengling Capsules.METHODS The analysis was performed on a 30℃thermostatic GL Science InertsilTM ODS-3 column(150 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,280 nm.Syringin was used as an internal standard to calculate the relative correction factors of the other nine constituents,after which the content determination was made.RESULTS Ten constituents showed good linear relationships within their own ranges(r≥0.999 7),whose average recoveries were 98.53%-102.22%with the RSDs of 1.26%-2.68%.The result obtained by QAMS approximated those obtained by external standard method.CONCLUSION This accurate and specific method can be used for the quality control of Tianma Toufengling Capsules.

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.

Objective:To investigate the application value of ultrasound technology in transurethral enucleation and resection of the prostate(TUERP).Methods:This study included 78 BPH patients admitted in our hospital from June 2021 to June 2023,aged 70.68±8.63 years and with the indication of surgery.We randomly divided them into two groups to receive TUERP(the control group,n=39)and ultrasound-assisted TUERP(the US-TUERP group,n=39).We statistically analyzed and compared the rele-vant parameters obtained before and after operation between the two groups.Results:No statistically significant differences were ob-served in the operation time and bladder irrigation time between the two groups(P>0.05).More glandular tissues were removed but less intraoperative bleeding and fewer perioperative complications occurred in the US-TUERP group than in the control.Compared with the baseline,IPSS,postvoid residual urine volume(PVR),quality of life score(QOL)and maximum urinary flow rate(Qmax)were significantly improved in both groups at 1 and 3 months after surgery,even more significantly in the US-TUERP than in the control group(P<0.05).Conclusion:US-TUERP helps achieve complete resection of the hyperplastic prostatic tissue along the surgical capsule at the anatomical level,with a higher safety,fewer perioperative complications,and better therapeutic effects.

Objective:To evaluate the safety and efficiency of China-made Toumai Robot-assisted laparoscopic radical prosta-tectomy(LRP).Methods:This study included 40 cases of PCa treated from January 2023 to May 2023 by robot-assisted LRP with preservation of the bladder neck and maximal functional urethral length,15 cases with the assistance of Toumai Robot(the TMR group)and the other 25 with the assistance of da Vinci Robot as controls(the DVR group).We recorded the docking time,laparo-scopic surgery time,vesico-urethral anastomosis time,intraoperative blood loss and postoperative urinary continence,and compared them between the two groups.Results:Operations were successfully completed in all the cases.No statistically significant differ-ences were observed between the TMR and DVR groups in the docking time(6 min vs 5 min,P＞0.05)or intraoperative blood loss(200 ml vs 150 ml,P＞0.05).The TMR group,compared with the DVR group,showed a significantly longer median laparoscopic surgery time(146 min vs 130 min,P＜0.05)and median vesico-urethral anastomosis time(19 min vs 16 min,P＜0.05).There were no statistically significant differences between the TMR and DVR groups in the rates of urinary continence recovery immediately af-ter surgery(60.0％[9/15]vs 64.0％[16/25],P＞0.05)or at 1 month(80.0％[12/15])vs(76.0％[19/25],P＞0.05),3 months(93.3％[14/15])vs(92.0％[23/25],P＞0.05)and 6 months postoperatively(100％[15/15])vs(96％[24/25],P＞0.05).Conclusion:China-made Toumai? Robot surgical system is safe and reliable for laparoscopic radical prosta-tectomy,with satisfactory postoperative recovery of urinary continence.

Objective:To explore the application of modified scrotoscopic surgery(MSS)in the treatment of testicular hydro-cele.Methods:We selected 45 cases of testicular hydrocele for this study,22 treated by traditional scrotoscopic surgery(TSS)and the other 23 by MSS,which was performed with a pin-shaped electrode bent inward at an angle of 60° instead of a circular electrode used in TSS.We recorded the general clinical data,operation time,incision length,intraoperative injury,incision infection,scrotal e-dema,postoperative hospital stay and postoperative Visual Analogue Scale(VAS)scores of the patients and compared them between the two groups.Results:There was no statistically significant difference in the general clinical data between the two groups(P＞0.05).Compared with the patients of the TSS group,those of the MSS group showed significantly shorter operative time([32.86±3.80]vs[26.13±2.81]min,P＜0.05),incision length([14.09±2.23]vs[8.73±1.48]mm,P＜0.05)and postopera-tive hospital stay([4.36±1.05]vs[2.00±0.90]d,P＜0.05),and achieved remarkably lower VAS scores on postoperative days 1(4.41±1.05 vs 3.09±0.79,P＜0.05),2(3.36±1.05 vs 2.78±1.13,P＜0.05),3(2.65±0.72 vs 1.74±0.86,P＜0.05)and 7(1.91±0.81 vs 1.04±0.82,P＜0.05).At 3 and 7 days after surgery,scrotal edema was markedly mil-der in the MSS than in the TSS group(P＜0.05).No testicular or epididymal damage,or wound infection occurred in either of the two groups.Conclusion:MSS is safe and effective in the treatment of testicular hydrocele,superior to TSS for its advantages of shorter operation time,smaller surgical incision,less postoperative pain and milder scrotal edema.

Objective To investigate the effect of Ganoderma lucidum polysaccharide peptide(GLPP)on proliferation,migration and apoptosis of diffuse large B cell lymphoma(DLBCL)cells and its mechanism.Methods OCI-LY19 cells were divided into six groups:control,GLPP,si-NC,si-protein arginine methyltransferase 6(PRMT6),GLPP+pcDNA3.1-NC and GLPP+pcDNA3.1-PRMT6 groups.The si-NC,si-PRMT6,GLPP+pcDNA3.1-NC and GLPP+pcDNA3.1-PRMT6 groups were transfected with si-NC,si-PRMT6,pcDNA3.1-NC and pcDNA3.1-PRMT6,respectively.After the transfection was completed,control,si-NC and si-PRMT6 groups were treated with RPMI-1640 medium,while the GLPP,GLPP+pcDNA3.1-NC and GLPP+pcDNA3.1-PRMT6 groups were cultured with RPMI-1640 medium containing with 20 μg·mL-1 GLPP.After administration 24 h,the cell proliferation inhibition rates,mobility rates and apoptosis rates were detected.The expression levels of PRMT6 protein were measured by Western blotting.Results The cell proliferation inhibition rates of si-NC,si-PRMT6,GLPP+pcDNA3.1-NC and GLPP+pcDNA3.1-PRMT6 groups were(1.28±0.16)％,(38.61±3.29)％,(52.84±7.74)％and(22.75±3.87)％,respectively.The number of cell migrations in the control,GLPP,si-NC,si-PRMT6,GLPP+pcDNA3.1-NC and GLPP+pcDNA3.1-PRMT6 groups was(252.65±24.65),(136.54±16.46),(231.65±21.24),(142.76±15.34),(140.23±9.84)and(192.38±23.38)cells;the apoptosis rates were(4.36±0.52)％,(28.24±2.36)％,(4.23±0.45)％,(24.54±2.27)％,(28.42±3.85)％and(14.25±2.13)％);the expression levels of PRMT6 protein were 1.82±0.21,0.56±0.05,1.78±0.19,0.54±0.05,0.29±0.02 and 0.32±0.03,respectively.The differences of above indexes were statistically significant between control group and GLPP group,between si-NC group and si-PRMT6 group,between GLPP+pcDNA3.1-NC group and GLPP+pcDNA3.1-PRMT6 group(all P＜0.05).Conclusion GLPP could inhibit proliferation,migration and promote apoptosis of DLBCL cells by down-regulating PRMT6 expression.

This study systematically analyzed the current status of outcomes in randomized controlled trial(RCT) of traditional Chinese medicine(TCM) treatment of diabetic kidney disease(DKD), aiming to provide a reference for constructing the core outcome set(COS) of TCM treatment of DKD. The clinical RCTs of TCM treatment of DKD that were published from January 2019 to March 2024 were retrieved from seven databases: CNKI, Wanfang, VIP, SinoMed, PubMed, Cochrane Library, and Web of Science. The risk of bias was assessed and outcome indicators were qualitatively analyzed. A total of 323 RCTs were included, reporting 199 outcome indicators with the frequency of 3 744 in seven indicator domains: TCM syndromes(196 times, 5.24%), symptoms and signs(275 times, 7.35%), physical and chemical examinations(2 820 times, 75.32%), safety events(422 times, 11.27%), long-term prognosis(21 times, 0.56%), quality of life(9 times, 0.24%), and economic evaluation(1 time, 0.02%). The three most frequently applied outcome indicators were total clinical response rate(268 times, 82.97%), serum creatinine(251 times, 77.71%), and urea nitrogen(206 times, 63.78%). The outcomes in the studies of TCM treatment of DKD had the limitations of high risk of bias, lack of standardization of TCM syndromes and efficacy evaluation, neglect of endpoint indicators, large differences in endpoint indicators and time points of measurement, unclear primary and secondary outcomes, few application of long-term prognosis, quality of life, and economic indicators, and insufficient reporting of safety events. In the future, efforts should be made to actively promote the construction of COS in the studies about the TCM treatment of DKD and enhance the quality of the clinical studies.
Humans ; Randomized Controlled Trials as Topic ; Diabetic Nephropathies/drug therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Treatment Outcome ; Medicine, Chinese Traditional ; Quality of Life