Objective:To assess the utility of whole-genome sequencing (WGS) in predicting Mycobacterium tuberculosis resistance to fluoroquinolones (FQs) and to establish a quantitative relationship between resistant gene mutations and resistance levels. Methods:A total of 296 drug-resistant tuberculosis surveillance strains with various resistance profiles, preserved by the National Tuberculosis Reference Laboratory of the Tuberculosis Prevention and Control Center at the Chinese Center for Disease Control and Prevention between 2013 and 2020, were included as study subjects. The Sensititre? MYCOTBI microplate method and WGS were used to assess the phenotypic and genotypic drug sensitivity of Mycobacterium tuberculosis to ofloxacin and moxifloxacin. Sensitivity, specificity, and concordance (Kappa value) of WGS in predicting fluoroquinolone sensitivity were calculated using phenotypic drug susceptibility testing (DST) results as the gold standard. A summary analysis was conducted on the distribution of drug resistance mutation sites and resistance levels. The paired χ 2 test was used to compare the detection rates between the two methods, with P<0.05 indicating statistical significance. Results:Among the 296 Mycobacterium tuberculosis strains with different resistance profiles, 196 were rifampicin-resistant, 50 were resistant to other drugs, and 50 were fully sensitive. WGS identified 81 strains carrying FQs resistance-related mutations, primarily at gyrA codons 94, 90, and 91. Sensitivity, specificity, and consistency (Kappa value) of WGS in predicting ofloxacin resistance were 86.5%, 98.1%, and 0.87, respectively. For moxifloxacin resistance prediction, these values were 80.0%, 99.5%, and 0.83, respectively. There was no statistically significant difference between the phenotypic DST and WGS detection rates for ofloxacin resistance (30.1% vs 27.4%, χ 2=3.06, P=0.08). However, the phenotypic DST detection rate for moxifloxacin resistance (33.8%, 100/296) was significantly higher than that of WGS (27.4%, 81/296) (χ 2=15.43, P<0.01). Analysis of the distribution of resistance mutation sites and resistance levels showed that different mutation sites corresponded to different minimum inhibitory concentrations (MICs). Multiple mutation combinations, including gyrA_D94G, gyrA_D94Y, and gyrA_D94N were mainly associated with high-level resistance, while gyrA_D94A, gyrA_A90V, and gyrA_S91P were primarily linked to low-level resistance. Conclusion:WGS demonstrates favorable sensitivity, specificity, and consistency in predicting FQs resistance and can partially predict resistance levels.

Cyclodipeptide (CDP) composed of two amino acids is the simplest cyclic peptide. These two amino acids form a typical diketopiperazine (DKP) ring by linking each other with peptide bonds. This characteristic stable ring skeleton is the foundation of CDP to display extensive and excellent bioactivities, which is beneficial for CDPs' pharmaceutical research and development. The natural CDP products are well isolated from actinomycetes. These bacteria can synthesize DKP backbones with nonribosomal peptide synthetase (NRPS) or cyclodipeptide synthase (CDPS). Moreover, actinomycetes could produce a variety of CDPs through different enzymatic modification. The presence of these abundant and diversified catalysis indicates that actinomycetes are promising microbial resource for exploring CDPs. This review summarized the pathways for DKP backbones biosynthesis and their post-modification mechanism in actinomycetes. The aim of this review was to accelerate the genome mining of CDPs and their isolation, purification and structure identification, and to facilitate revealing the biosynthesis mechanism of novel CDPs as well as their synthetic biology design.
Actinobacteria/metabolism* ; Actinomyces/metabolism* ; Biological Products/metabolism* ; Bacteria/metabolism* ; Diketopiperazines/metabolism* ; Amino Acids

Objective:To evaluate the efficacy and safety of mitoxantrone hydrochloride liposome injection in the treatment of peripheral T-cell lymphoma (PTCL) in a real-world setting.Methods:This was a real-world ambispective cohort study (MOMENT study) (Chinese clinical trial registry number: ChiCTR2200062067). Clinical data were collected from 198 patients who received mitoxantrone hydrochloride liposome injection as monotherapy or combination therapy at 37 hospitals from January 2022 to January 2023, including 166 patients in the retrospective cohort and 32 patients in the prospective cohort; 10 patients in the treatment-na?ve group and 188 patients in the relapsed/refractory group. Clinical characteristics, efficacy and adverse events were summarized, and the overall survival (OS) and progression-free survival (PFS) were analyzed.Results:All 198 patients were treated with mitoxantrone hydrochloride liposome injection for a median of 3 cycles (range 1-7 cycles); 28 cases were treated with mitoxantrone hydrochloride liposome injection as monotherapy, and 170 cases were treated with the combination regimen. Among 188 relapsed/refractory patients, 45 cases (23.9%) were in complete remission (CR), 82 cases (43.6%) were in partial remission (PR), and 28 cases (14.9%) were in disease stabilization (SD), and 33 cases (17.6%) were in disease progression (PD), with an objective remission rate (ORR) of 67.6% (127/188). Among 10 treatment-na?ve patients, 4 cases (40.0%) were in CR, 5 cases (50.0%) were in PR, and 1 case (10.0%) was in PD, with an ORR of 90.0% (9/10). The median follow-up time was 2.9 months (95% CI 2.4-3.7 months), and the median PFS and OS of patients in relapsed/refractory and treatment-na?ve groups were not reached. In relapsed/refractory patients, the difference in ORR between patients with different number of treatment lines of mitoxantrone hydrochloride liposome injection [ORR of the second-line, the third-line and ≥the forth-line treatment was 74.4% (67/90), 73.9% (34/46) and 50.0% (26/52)] was statistically significant ( P = 0.008). Of the 198 PTCL patients, 182 cases (91.9%) experienced at least 1 time of treatment-related adverse events, and the incidence rate of ≥grade 3 adverse events was 66.7% (132/198), which was mainly characterized by hematologic adverse events. The ≥ grade 3 hematologic adverse events mainly included decreased lymphocyte count, decreased neutrophil count, decreased white blood cell count, and anemia; non-hematologic adverse events were mostly grade 1-2, mainly including pigmentation disorders and upper respiratory tract infection. Conclusions:The use of mitoxantrone hydrochloride liposome injection-containing regimen in the treatment of PTCL has definite efficacy and is well tolerated, and it is a new therapeutic option for PTCL patients.

Objective:To investigate the efficacy of 3D-printed quantitative bone implants assisting second-stage Masquelet technique for the treatment of long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures.Methods:A retrospective case series analysis was made on 26 patients with long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures treated in Wuxi Ninth People′s Hospital from July 2015 to December 2020, including 20 males and 6 females; aged 19-63 years [(46.5±4.5)years]. Gustilo classification was type IIIB in 23 patients and type IIIC in 3. In the first stage, all patients had thoroughly emergent debridement, removal of all free bone pieces, restoration of the length and force line plus externally fixion, and vacuum sealing drainage (VSD) of the residual wound. After 2-7 days, the external fixation was removed and replaced by internal fixation, with the bone cement filling in the defect area and the free flap covering the wound. The length of tibial bone defect was 5-14 cm [(6.3±0.4)cm], and the tibial defect volume was 12.2-73.1 cm 3 [(33.6±9.2)cm 3]. In the second stage (6-19 weeks after injury), the bone cement was removed, followed by autologous bone grafting. Prior to bone grafting, digital technology was used to accurately calculate the bone defect volume, and an equal volume of bone harvesting area was designe to produce the 3D printed osteotomy template. Bone grafting was conducted after bone removal according to the osteotomy template during operation. The success rate of one-time iliac bone extraction, bone harvesting time, and bleeding volume were recorded. Pain in the bone extraction area was evaluated by visual analogue score (VAS) at 1 day and 1 month after operation and at the last follow-up. Wound healing, complications, and bone healing were observed. Life quality was evaluated by health survey brief form (SF-36) including scores of physical component summary (PCS) and mental component summary (MCS) before bone grafting and at the last follow-up. Results:All the patients were followed up for 13-53 months [(32.3±12.5)months]. One-time iliac bone extraction was successful in all the patients. Bone harvesting time was 15-30 minutes [(21.0±2.5)minutes]. The bleeding volume was 50-120 ml [(62.3±29.0)ml]. The VAS was 1-4 points [(1.2±0.9)points] at 1 day after operation, higher than these (0.0±0.0)points at 1 month after operation and at the last follow-up (all P<0.01). Totally, 25 patients obtained wound healing after operation, except for 1 patient with superficial wound infection after bone grafting that was healed by dressing change. There was 1 patient with bone infection after 3 months of bone grafting that was healed by repeated surgery with Masquelet technique in the first and second stage. Besides, 2 patients had symptoms of cutaneous nerve injury in the iliac donor area. The time of bone healing was 4-7 months [(5.8±0.8)months]. The scores of PCS and MCS in SF-36 at the last follow-up were (73.6±12.8)points and (83.6±13.2)points, significantly higher than those before bone grafting [(46.8±0.5)points, (60.7±2.0)points] (all P<0.01). Conclusion:Second-stage Masquelet technique with 3D printed quantitative bone implants for the treatment of long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures is associated with shortened bone harvesting time, attenuated pain, reduced complications, accelerated bone healing and improved function.

Objective:To compare the efficacy of concurrent and asynchronous radiochemotheray for early extranodal nasal natural killer/T-cell lymphoma (NKTCL).Methods:From 2007 to 2020, 278 patients with early NKTCL treated with comprehensive treatment in the Affiliated Tumor Hospital of Guizhou Medical University were recruited. According to the adjusted Nomogram-revised risk index (NRI) prognostic model, there were 49 cases in the good prognostic group without adverse prognostic factors (age>60 years old, increased serum lactate dehydrogenase (LDH), ECOG score ≥2, primary tumor invasion (PTI), Ann Arbor stage Ⅱ, and 229 cases in the poor prognostic group with any adverse prognostic factors. 145 of these cases were treated with concurrent radiochemotherapy, and 133 of them were treated with asynchronous radiochemotherapy.Results:The 5-year overall survival (OS) rate of the whole group was 71.0%, and the progression-free survival (PFS) rate was 67.6%. The 5-year OS rate in the good prognostic group was 95.6%, and 65.4% in the poor prognostic group ( P<0.001). In the poor prognostic group, the 5-year OS rates of patients with NRI=1(low-and moderate-risk group), NRI=2(moderate-and high-risk group), NRI≥3(high-risk group) were 72.1%, 61.1% and 47.7%, respectively ( P=0.007). There was no significant difference in curative effect between the concurrent and asynchronous radiochemotherapy groups. The 5-year OS rates were 70.6% and 69.8%( P=0.783), and the 5-year PFS rates were 67.6% and 65.2%( P=0.631). Further stratified analysis showed that the 5-year OS rates of patients with NRI=1 receiving concurrent and asynchronous radiochemotherapy were 73.1% and 76.5%( P=0.576), 62.6% and 69.3%( P=0.427) for those with NRI=2, and 58.1% and 42.3% for those with NRI≥3( P=0.954). Conclusions:Comprehensive treatment can significantly improve the prognosis of early NKTCL in the poor prognostic group. In the sequence of radiotherapy and chemotherapy, there is no significant difference in 5-year OS and PFS rates between concurrent and asynchronous radiochemotherapy. Sequential treatment with better tolerance can be adopted for early NKTCL with poor prognosis.

β-thalassemia is a single-gene genetic disease caused by β globin gene mutations leading to the fact that red blood cells are unable to form normal adult hemoglobin, and then patients develop hemolytic anemia. Current treatment regimens mainly include allogenetic hematologic stem cell transplantation, symptomatic regular blood transfusions and the use of iron removers to reduce iron load. Some severe patients have quite poor prognoses and deadly consequences if not treated timely. Genetically modified autohematopoietic stem cells can provide a new treatment option for patients with β thalassemia, which may achieve a long-term and stable increase in hemoglobin level through a single dose, making one-time cure β-thalassemia possible. This paper reviews the key elements of clinical trial design for β-thalassemia gene therapy from the aspects of efficacy evaluation endpoints, clinical trial design, enrollment population, and subject monitoring in order to provide a reference for pharma-therapeutic research and development enterprises.

Objective To investigate the pollution levels of six indicator polychlorinated biphenyls (PCBs) in animal-derived foods in Hebei Province. Methods A total of 96 food samples (including beltfish, snakehead, egg, pork liver, pork and beef meat) were collected from Hebei Province in 2019. The contents of six indicator PCBs (PCB 28, PCB 52, PCB 101, PCB 138, PCB 153 and PCB180) in different food samples were determined using isotope dilution-high resolution gas chromatography-high resolution mass spectrometry (HRGC-HRMS). Results The contents of indicative PCBs in fish samples ranged from 0.12 to 0.32 ngg-1 fresh weight, while the content of PCBs in other samples ranged from 0.11 to 1.10 ng g-1 fat. PCB 138 and PCB 153 were the main compounds detected in fish samples, while PCB 28 and PCB 52 were the main compounds detected in other samples. The average estimated daily dietary intake of PCBs for residents in Hebei Province was 0.14 ng kg^-1·bw^-1·d^-1, and the highest contribution rate was beltfish (61.52%). Conclusion The pollution levels of indicator PCBs in animal-derived foods in Hebei Province are relatively low, and the residents has lower health risks through dietary intake of indicator PCBs.

Objective:To compare the clinical effect of buttress plate fixation and cannulated screw fixation of Regan-Morrey type II ulnar coronoid fractures.Methods:A retrospective case control study was made on 53 patients with Regan-Morrey type II ulnar coronoid fractures admitted to Wuxi No.9 People 's Hospital from April 2015 to January 2018，including 36 males and 17 females，aged from 21 to 63 years［（36.3±7.1）years］. Among them，24 patients were treated using buttress plates（plate group），and 29 patients using cannulated screws（screw group）. The operation time and fracture healing time were documented. The visual analogue score（VAS），range of motion of elbow flexion and extension and forearm rotation and Mayo elbow performance score（MEPS）were assessed at postoperative 1，3，6 months and at the last follow-up. The incidence of complications was observed as well. Results:All patients were followed up for 15-18 months［（15.9±1.3）months］. The operation time in plate group［（150.6±24.2）minutes］was longer than that in screw group［（126.8±18.3）minutes］（ P<0.05）. There was no significant difference in fracture healing time or VAS between the two groups（ P>0.05）. After 1，3，6 months and during the last follow-up，the range of motion of elbow joint flexion and extension in plate group［（87.2±5.8）°，（109.2±7.1）°，（121.3±6.2）°，（127.3±5.4）°］was higher than that in screw group［（70.5±9.1）°，（90.2±4.5）°，（108.3±5.1）°，（116.2±4.6）°］，the range of motion of forearm rotation in plate group［（78.3±9.1）°，（98.7±8.6）°，（130.2±7.1）°，（139.2±6.7）°］was higher than that in screw group［（60.1±5.1）°，（80.6±8.7）°，（116.1±5.5）°，（127.3±4.1）°］，and the MEPS in plate group［（30.6±7.6）points，（68.1±6.1）points，（90.2±4.3）points，（95.2±2.1）points］was higher than that in screw group［（27.2±8.1）points，（54.1±7.1）points，（82.1±5.3）points，（88.2±5.2）points］（all P<0.05）. In plate group，one patient sustained superficial wound infection at postoperative 1 week，which was healed uneventfully after surgical debridement and antibiotic therapy；two patients had heterotopic ossification without addition surgery. In screw group，three patients presented screw loosening and fracture redisplacement during early movement，which was healed by reducing the intensity of elbow functional exercise and prolonging the protection time of brace；four patients had heterotopic ossification，among which one combined with elbow stiffness showed improved range of motion of the elbow after elbow release at postoperative 12 months. The incidence of complications in plate group［13%（3/24）］was lower than that in screw group［26%（7/29）］（ P<0.05）. Conclusion:For Regan-Morrey type II ulnar coronoid fractures，the buttress plate fixation is superior to the cannulated screw fixation in fixation strength，recovery of elbow function and incidence of complications in regardless of longer operation time.

Objective:To analyze the efficacy and prognostic factors of radiotherapy combined with asparaginase/peaspartase-based chemotherapy regimen in the treatment of early stage extranodal natural-killer/T cell lymphoma of the upper aerodigestive tract (UADT ENKTCL).Methods:267 early stage UADT ENKTCL patients were treated in Guizhou Cancer Hospital from October 2003 to February 2020. Among them, 229 patients received radiotherapy or radiotherapy combined with menpartaminase/permenidase-based chemotherapy regimen and 38 patients were treated with radiotherapy or chemotherapy alone. The overall survival (OS) and progression-free survival (PFS) were calculated by Kaplan- Meier method, log-rank test was conducted for univariate analysis and Cox regression model was performed for multivariate analysis. Results:The 5-year OS and PFS were 67.2% and 61.5% in all patients. The 5-year OS and PFS in patients treated with radiotherapy combined with chemotherapy, radiotherapy alone and chemotherapy alone were 71.7%, 35% and 49%(all P<0.001), and 66.4%, 35% and 28%(all P<0.001), respectively. According to the NRI risk stratification, 246 patients treated with radiotherapy and chemotherapy were divided into the favourable and the unfavourable prognosis groups. The 5-year OS was 93.3% and 64.3%( P<0.001) and the 5-year PFS was 91.1% and 56.7%( P<0.001) in two groups. For patients receiving radiotherapy with a dose ≥50 Gy and<50 Gy, the 5-year OS was 72.4% and 55.7%( P<0.001), and the 5-year PFS was 68.3%, and 36.5%( P<0.001). In the unfavourable prognosis group, the 5-year OS of patients receiving ≥ 4 and<4 cycles of chemotherapy was 65.5% and 59.2%( P=0.049), and the 5-year PFS was 60.7% and 50.6%( P=0.018). Univariate analysis showed that stage Ⅱ, ECOG≥2, primary tumor invasion, radiotherapy alone, NRI≥1(Nomogram-revised risk index), EBV-DNA≥2 750 copies/ml, radiotherapy dose < 50 Gy, and<4 cycles of chemotherapy were associated with unfavorable 5-year OS and PFS (all P<0.05), and CHOP-like regimen was the risk factor of unfavorable 5-year PFS ( P<0.05). Multivariate analysis demonstrated that primary tumor invasion, ECOG≥2, and radiotherapy dose <50 Gy were associated with unfavorable OS and PFS (all P<0.05), and stage Ⅱ was the risk factor of unfavorable 5-year OS ( P<0.05). Conclusions:The prognosis of early stage low-risk UADT ENKTCL of is favourable. Sufficient dose of extended involved-field radiotherapy is an important curative modality in early stage UADT ENKTCL. Compared with radiotherapy alone, radiotherapy combined with chemotherapy can significantly improve the prognosis of early stage UADT ENKTCL patients in the unfavourable prognosis group. Full-course chemotherapy can significantly prolong the long-term survival in the unfavorable prognosis group. The chemotherapy containing asparaginase can significantly enhance the prognosis of patients with early stage UADT ENKTCL.

Objective:To analyze the efficacy and prognostic factors of induced chemotherapy combined with radiotherapy in the treatment of early stage extranodal natural-killer/T cell lymphoma (ENKTCL).Methods:Two hundred and eighty-seven early stage NKTCL patients were treated in Affiliated Cancer Hospital of Guizhou Medical University from October 2003 to October 2021. All patients were aD ministrated with short courses of induced chemotherapy combined with radiotherapy. Clinical prognostic factors of early stage NKTCL were analyzed. The overall survival (OS) and progression-free survival (PFS) were calculated by Kaplan- Meier method, log-rank test was conducted for univariate analysis and Cox models were performed for multivariate analysis. Results:The 5-year OS and PFS were 72.8% and 68.9% in all patients. According to Nomogram risk index (NRI) prognostic model, 286 patients were divided into the low risk (NRI=0), intermediate low risk (NRI=1), intermediate high risk (NRI=2), high risk (NRI=3) and very high risk (NRI≥4) groups. In these 5 groups, the 5-year OS were 95.6%, 76.3%, 69.5%, 61.0% and 23.3%(all P<0.001), and the 5-year PFS were 93, 2%, 69.8%, 64.6%, 60.2% and 23.3%(all P<0.001), respectively. In the radiotherapy with a dose of ≥50 Gy and<50 Gy groups, the 5-year OS were 73.8% and 65.9%( P=0.123) and the 5-year PFS were 72.8% and 45.3%( P=0.001). According to the response to induced chemotherapy of complete response (CR), partial response (PR), stable disease (SD) and progressive disease (PD), the 5-year OS were 85.4%, 74.0%, 61.8% and 28.5%(all P<0.001), and the 5-year PFS were 83.7%, 66.8%, 65.7% and 27.4%(all P<0.001), respectively. Univariate analyses showed that stage Ⅱ, ECOG≥2, primary tumor invasion, radiotherapy dose<50 Gy and short-term efficacy of induced chemotherapy were poor prognostic factors for the 5-year OS and PFS (all P<0.05). Multivariate analyses demonstrated that primary tumor invasion, ECOG≥2 and stage Ⅱ were poor prognostic factors for OS (all P<0.05), and primary tumor invasion and ECOG≥2 were poor prognostic factors for PFS (all P<0.05). Conclusions:Early stage NKTCL patients can obtain high efficacy after induced chemotherapy combined with radiotherapy. Complete response to induced chemotherapy is associated with favorable prognosis.