The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26％of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32％and 110.69％of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.

The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26％of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32％and 110.69％of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.

As one of the key components of clinical trials, blinding, if successfully implemented, can help to mitigate the risks of implementation bias and measurement bias, consequently improving the validity and reliability of the trial results. However, successful blinding in clinical trials of traditional Chinese medicine (TCM) is hard to achieve, and the evaluation of blinding success through blinding assessment lacks established guidelines. Taking into account the challenges associated with blinding in the TCM field, here we present a framework for assessing blinding. Further, this study proposes a blinding assessment protocol for TCM clinical trials, building upon the framework and the existing methods. An assessment report checklist and an approach for evaluating the assessment results are presented based on the proposed protocol. It is anticipated that these improvements to blinding assessment will generate greater awareness among researchers, facilitate the standardization of blinding, and augment the blinding effectiveness. The use of this blinding assessment may further advance the quality and precision of TCM clinical trials and improve the accuracy of the trial results. The blinding assessment protocol will undergo continued optimization and refinement, drawing upon expert consensus and experience derived from clinical trials. Please cite this article as: Wang XC, Liu XY, Shi KL, Meng QG, Yu YF, Wang SY, Wang J, Qu C, Lei C, Yu XP. Blinding assessment in clinical trials of traditional Chinese medicine: Exploratory principles and protocol. J Integr Med. 2023; 21(6): 528-536.
Drugs, Chinese Herbal/therapeutic use* ; Medicine, Chinese Traditional/methods* ; Outcome Assessment, Health Care ; Reference Standards ; Reproducibility of Results ; Research Design ; Clinical Trials as Topic

Objective:To explore the risk factors of recurrence after resection of thigh soft tissue malignant tumors and the evaluation of the postoperative lower limb function.Methods:A total of 211 patients who underwent thigh soft tissue malignant tumor resection in the Department of Orthopaedics of the second affiliated Hospital of Medical College of Zhejiang University from May 2011 to May 2021 were retrospectively analyzed, including 117 males and 94 females, aged 53 (43, 65) years. There were 59 cases of atypical/well-differentiated liposarcoma, 30 cases of other types of liposarcoma, 33 cases of malignant fibrous histiocytoma, 19 cases of fibrosarcoma, 12 cases of rhabdomyosarcoma, 12 cases of leiomyosarcoma, 9 cases of synovial sarcoma, and 37 cases of others. The tumor involvement of muscle, bone, blood vessel, nerve tissue and intermuscular compartment, postoperative Musculoskeletal Tumor Society (MSTS) functional score and recurrence were recorded. The relationship between anatomical structure and postoperative recurrence was analyzed by Cox logistic regression to determine risk factors; the cumulative recurrence rate of different compartment groups (medial compartment, anterior compartment, and posterior compartment) were compared; The Box plot was used to compare the postoperative function of patients with resection of different anatomical structures (e.g., vastus lateralis, biceps femoris, femur, etc.).Results:A total of 34 out of 211 patients relapsed, with a recurrence rate of 16.1%. The recurrence time ranged from 2.6 months to 91.6 months after operation, with a median recurrence time of 37.0 (18.2, 52.8) months. Three independent risk factors were found to be associated with recurrence, namely: pathological grade [ HR=3.86, 95% CI(1.75, 8.51)], involvement of vastus intermedius [ HR=3.05, 95% CI(1.53, 6.06)], and involvement of vastus medialis [ HR=3.17, 95% CI(1.56, 6.41)]. The recurrence rate of patients with anterior chamber tumor resection was 35.3%, which was higher than that of patients without tumor resection (16.2%), and the difference was statistically significant ( P=0.020). There was no significant difference in recurrence rate between patients with medial chamber tumor resection and patients without tumor resection ( P>0.05). The recurrence rate of patients with posterior compartment tumor resection was 12.3%, which was lower than that of unresectable patients (37.6%), and the difference was statistically significant ( P=0.002).The postoperative MSTS score of 167 patients averaged 26±3.2 points (9-30 points). After intraoperative resection of part of the femur, vastus intermedius, vastus medialis, and rectus femoris, the patient's function was relatively poor [corresponding median MSTS score: 25 (23, 28), 26 (24 28), 26 (24,28), 26(24, 27)]. Conclusion:The risk factors for local recurrence after resection of thigh soft tissue malignant tumors include: pathological grade of the tumor, and whether the vastus intermedius or vastus medialis is involved. Anterior compartment tumors have a higher recurrence rate after surgery. If the tumor involves the above-mentioned anatomical structures, more attention should be paid to the risk of local recurrence after surgery.

Objective:To explore the clinical characteristics of dislocation after resection and reconstruction of tumors involving pelvic area II, and to try to propose intraoperative and postoperative techniques to prevent its occurrence.Methods:From March 2011 to March 2021, 122 patients with resection and reconstruction involving pelvic area II were retrospectively analyzed. Among them, 17 had postoperative dislocation, 32 had pelvic area II, and 31 had pelvic area I+II. There were 40 cases in the pelvic area II+III, and 19 cases in the pelvic area I+II+III. There were 49 female patients and 73 male patients; the mean age was 47 years (9-73 years). The Musculoskeletal Tumor Society (MSTS) score of lower extremity patients after reduction was evaluated, and the clinical characteristics of dislocation, such as dislocation direction, dislocation time and reduction method, were counted and analyzed, and feasible prevention measures were analyzed based on the imaging characteristics after resection and reconstruction. dislocation method.Results:Among the 122 patients, there were 17 cases of dislocation, and the dislocation rate was 13.7%. Among them, 12 cases were anterior dislocation, accounting for 70.6% of anterior dislocation; 5 cases were posterior dislocation, and the proportion of posterior dislocation was 29.4%, difference (χ 2=4.52, P=0.033). There were 12 cases of dislocation within 3 months after operation, accounting for 70.6%; 1 case of dislocation occurred in 5 months after operation, and the other 4 cases of dislocation occurred for more than one year. The MSTS score of the dislocation patients after reduction was 56.1±15.6% (20%-80%). Combined with postoperative imaging examinations, the feasible methods for preventing dislocation include: The anteversion angle of the acetabular cup can be appropriately reduced (0°-10°); The acetabular cup can be appropriately shifted to the rear of the rotation center; Appropriately reducing the anteversion angle of the femoral neck prosthesis, the combination of the three can reduce the occurrence of femoral-acetabular prosthesis impingement; at the same time, the affected limb is controlled in a neutral position with strict nail shoes after surgery to avoid the occurrence of external rotation. Conclusion:The dislocation after tumor resection and reconstruction in pelvic area II is mostly anterior dislocation. Postoperative CT scan of the pelvis shows that the space between the neck of the femoral stem prosthesis and the lower edge of the acetabulum is small, which is prone to impingement. If the anteversion angle of the acetabular cup is appropriately reduced, the acetabular prosthesis is appropriately displaced posteriorly, and the anteversion angle of the femoral neck is reduced by 5°, the possibility of femoral-acetabular impingement can be reduced, thereby reducing the risk of postoperative dislocation.

Objective:To explore the cognition of community health care workers and the elderly on frailty, and attitudes towards frailty management.Methods:The semi-structured interview was conducted with 14 community health care workers and 17 community elderly in 3 communities in Beijing with the method of phenomenological research. We interviewed respondents' cognition on frailty management and attitudes towards frailty management. Colaizzi 7-step method was used for data analysis.Results:Three themes of cognition on elderly frailty were extracted including that the elderly in the community had different cognition and understandings of elderly frailty; community health care workers had a comprehensive understanding of the concept of elderly frailty, but not much contact with this concept; there were several misunderstandings about the cognition of elderly frailty. Four themes of attitudes towards frailty management were extracted including that the elderly want scientific frailty management; the elderly in the early stage of frailty did not agree with frailty management; community health care workers affirmed the importance of frail management; there were many obstacles to the frailty management in Beijing communities.Conclusions:It is necessary to strengthen education and publicity for the elderly frailty, speed up the formulation of evidence-based frailty management plans, guide community practice, and improve the quality of life of the elderly.

BACKGROUNDProteasome subunits (PSMB) and transporter associated with antigen processing (TAP) loci are located in the human leukocyte antigen (HLA) Class II region play important roles in immune response and protein degradation in neurodegenerative diseases. This study aimed to explore the association between single nucleotide polymorphisms (SNPs) of PSMB and TAP and Parkinson's disease (PD).

METHODSA case-control study was conducted by genotyping SNPs in PSMB8, PSMB9, TAP1, and TAP2 genes in the Chinese population. Subjects included 542 sporadic patients with PD and 674 healthy controls. Nine identified SNPs in PSMB8, PSMB9, TAP1, and TAP2 were genotyped through SNaPshot testing.

RESULTSThe stratified analysis of rs17587 was specially performed on gender. Data revealed that female patients carry a higher frequency of rs17587-G/G versus (A/A + G/A) compared with controls. But there was no significant difference with respect to the genotypic frequencies of the SNPs in PSMB8, TAP1, and TAP2 loci in PD patients.

CONCLUSIONChinese females carrying the rs17587-G/G genotype in PSMB9 may increase a higher risk for PD, but no linkage was found between other SNPs in HLA Class II region and PD.

ATP-Binding Cassette Sub-Family B Member 2 ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; genetics ; Adult ; Aged ; Antigen Presentation ; Case-Control Studies ; Cysteine Endopeptidases ; genetics ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; immunology ; Polymorphism, Single Nucleotide ; Proteasome Endopeptidase Complex ; genetics

Objective:To evaluate the protective immunity by vaccination of BALB/c mice with rLV-HA-GCN4,a recombinant lentivirus expressing the trimeric HA of swine H1N1 influenza virus. Methods:The female mice were randomly divided into rLV-HA-GCN4,rLV-HA,LV and PBS groups. Mice were primed with plasmid and boosted with lentivirus by the administration of intramuscular thigh injections at an interval of two weeks. At day 28 post-prime immunization,mice were inoculated intranasally with 100TCID50 of swine H1N1 influenza virus in a 50 μl volume. The immune levels were assessed by the T lymphocyte transformation test, flow cytometry,indirect ELISA and the indexes of spleen and lung. Results:The spleen lymphocyte transformation rate was 0. 3±0. 11 in the rLV-HA-GCN4 group at day 14 post-boost immunization, showing a statistical significance ( P<0. 01 ) compared to the PBS group. Meanwhile,rLV-HA-GCN4 could cause T lymphocyte response mainly based on the Th1-type CD4+ T cells. The IgG antibody titer reached to 1:8 000 at day 14 post-boost immunization and approximately 1:7 000 at day 14 post challenge. After challenge,the spleen and lung indexes of rLV-HA-GCN4 group were significantly lower than those of PBS group (P<0. 05). The body weight of rLV-HA-GCN4 group demonstrated a slight decrease before 3 days post challenge and then a gradual increase compared to the LV and PBS groups (P<0. 05). Conclusion:rLV-HA-GCN4 can effectively induce cellular and humoral immune response in BALB/c mice against swine H1N1 influenza virus.

Objective To compare the effect of titanium elastic nailing (TEN) and plating in treatment of both radius and ulna fractures in children older than 10 years.Methods This retrospective analysis comprised 38 children (50 sides) of both radius and ulna fractures treated with TEN (30sides) or plate (20 sides) between 2009 and 2014.Parameters compared between the two fixation methods were operation time,ratio of open/close reduction,time to internal fixation removal,incidence of postoperative complications,and bone union time.Anderson score was used to evaluate the treatment results.Results Operation time [(94.3 ± 39.5) min vs (120.5 ± 45.4) min],time to internal fixation removal [(210.8 ± 84.1) d vs (264.4 ± 88.9) d],and ratio of open/close reduction (20/10 vs 20/0)varied significantly between TEN and plating groups (P ＜ 0.05 or 0.01).Whereas,no statistically significant differences were noted in regard to complication rate,bone union time,and effect evaluation.Conclusion For fracture of the radius and ulna in children older than 10 years,TEN should be used for closed type without obvious displacement,but plate fixation is associated with lower incidence of fracture malunion and better outcome in treatment of open type with obvious displacement.

OBJECTIVETo explore the technique and therapeutic effect of bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral shaft fractures in young children. Methods:From June 2006 to June 2012,45 young children with femoral shaft fractures were treated by bridge wire splint fixation with ankle dorsiflexion,which was designed according to arch bridge mechanical principle and structure. There were 31 males and 14 females with an average age of 3.2 years old ranging from 8 months to 5.5 years old; 14 cases were upper 1/3 femoral fractures,26 cases were middle 1/3 femoral fractures,5 cases were lower 1/3 femoral fractures; 20 cases were transverse fractures, 14 cases were oblique fractures,6 cases were spiral frac- tures, and 5 cases were comminuted fractures. X-ray, follow-up imaging changes,clinical curative effect and complications were assessed.

RESULTSForty-five patients were followed up for 6 to 21 months (averaged 12 months). All fractures were reached clinical bone healing after 5 to 7 weeks (averaged 6 weeks) fixation. Seven cases appearred limb soft tissue complications, including buttocks bedsore,dorsal foot and Achilles tendon epidermal necrosis, and healed after dressing and removal of external fixation. During follow-up,the original overlap angle and lateral displacement were remodeled, and limbs were restored to the normal line of force and bone structure. According to Flynn standard, 35 cases got excellent results, 8 cases good, 2 cases fair.

CONCLUSIONThe bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral. shaft fractures in young children (less than 6 years old) is safe,feasible, simple,and has raliable effect, which can be applied in primary hospitals.

Ankle ; surgery ; Bone Wires ; Child ; Child, Preschool ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; instrumentation ; Humans ; Infant ; Male ; Treatment Outcome