1.Mechanisms of Mycobacterium tuberculosis-induced primary cilia degra-dation in monocytes-macrophages and promotion of osteoclast differentia-tion
Chengran ZHANG ; Qianfei LIU ; Nanjun XIONG ; Junbao CHEN ; Qile GAO ; Mingxing TANG ; Shaohua LIU
Chinese Journal of Infection Control 2025;24(9):1208-1214
Objective To explore the effect of Mycobacterium tuberculosis infection on the primary cilia of mono-cytse-macrophages and its potential mechanisms of promoting osteoclast differentiation.Methods Bone marrow-de-rived mononuclear cells(BMMCs)isolated from patients in control group and spinal tuberculosis group(TB group)were performed in vitro culture,and then cultured with Mycobacterium tuberculosis,infection model(Rv group)was constructed.Changes in cilia were observed by fluorescence staining and scanning electron microscopy tech-nique,a mouse spinal TB model was constructed for validating.Results Compared with the control group,the ex-pression of primary cili markers in the lesion of bone tissue of patients in TB group decreased significantly;After co-culturing with Mycobacterium tuberculosis,the ratio(48.56%±7.77%vs 9.58%±5.59%)and length(4.050[3.289,4.666]μm vs 0[0,0.676]μm)of primary cilia of monocytes-macrophages in the Rv group decreased sig-nificantly;The infiltration of osteoclasts in the bone marrow cavity of spinal TB mice was obvious,and the propor-tion and length of primary cilia decreased significantly.Conclusion Intracellular infection of Mycobacterium tuberculosis can induce degradation of primary cilia in monocytes-macrophages,promote osteoclast differentiation,and exacerbate vertebral bone resorption.
2.Mechanisms of Mycobacterium tuberculosis-induced primary cilia degra-dation in monocytes-macrophages and promotion of osteoclast differentia-tion
Chengran ZHANG ; Qianfei LIU ; Nanjun XIONG ; Junbao CHEN ; Qile GAO ; Mingxing TANG ; Shaohua LIU
Chinese Journal of Infection Control 2025;24(9):1208-1214
Objective To explore the effect of Mycobacterium tuberculosis infection on the primary cilia of mono-cytse-macrophages and its potential mechanisms of promoting osteoclast differentiation.Methods Bone marrow-de-rived mononuclear cells(BMMCs)isolated from patients in control group and spinal tuberculosis group(TB group)were performed in vitro culture,and then cultured with Mycobacterium tuberculosis,infection model(Rv group)was constructed.Changes in cilia were observed by fluorescence staining and scanning electron microscopy tech-nique,a mouse spinal TB model was constructed for validating.Results Compared with the control group,the ex-pression of primary cili markers in the lesion of bone tissue of patients in TB group decreased significantly;After co-culturing with Mycobacterium tuberculosis,the ratio(48.56%±7.77%vs 9.58%±5.59%)and length(4.050[3.289,4.666]μm vs 0[0,0.676]μm)of primary cilia of monocytes-macrophages in the Rv group decreased sig-nificantly;The infiltration of osteoclasts in the bone marrow cavity of spinal TB mice was obvious,and the propor-tion and length of primary cilia decreased significantly.Conclusion Intracellular infection of Mycobacterium tuberculosis can induce degradation of primary cilia in monocytes-macrophages,promote osteoclast differentiation,and exacerbate vertebral bone resorption.
3.Initial dose and safety of cadmium-antidote GMDTC for intravenous infusion
Qile ZHAO ; Yuting GAO ; Wei HU ; Zhiyong ZHONG ; Xuefeng REN ; Xiaojiang TANG
China Occupational Medicine 2024;51(3):257-264
Objective To investigate the initial dose and safety of intravenous infusion of sodium (s)-2-(dithiocarboxylato((2R,3R,4R,5R,6R)-2,3,4,5,6-pentahydroxyhexyl) amino)-4-(methylthio) butanoate (GMDTC) for the displacement of cadmium. Methodsi) Efficacy test. The New Zealand male rabbits were randomly divided into model group, calcium disodium edetate (EDTA) group and GMDTC low-, medium- and high-dose groups after cadmium poisoning using 2.5 cadmium chloride dihydrate. Rabbits in EDTA group were intravenously injected with EDTA dipotassium at a dose of 93.5 mg/kg body weight, rabbits in the three doses groups were intravenously injected of GMDTC at doses of 12.0, 36.0, and 108.0 mg/kg body weight, respectively. The rabbits in the control group (separate set) and model group were intravenously injected with equal volumes of 0.9% sodium chloride solution, administered for five consecutive days per week for 1, 2, and 4 weeks. ii) Toxicity test. Specific pathogen free SD rats were randomly divided into solvent control group and low-, medium- and high-dose groups. In the acute toxicity test, the rats in the three-dose groups were intravenously injected of GMDTC at doses of 200.0, 800.0 and 3 000.0 mg/kg body weight, respectively. In the long-term toxicity test, the rats in the three-dose groups were intravenously injected GMDTC at doses of 100.0, 500.0 and 2 000.0 mg/kg body weight, respectively, once a day for four consecutive weeks, with a recovery period of four weeks. The rats in the solvent control group were given an equal volume 0.9% sodium chloride solution intravenously at the same time. The maximum tolerated dose (MTD) and no observable adverse effect level (NOAEL) were detected. Resultsi) In the one week treatment experiment, the 24 hours urinary cadmium levels of rabbits in the three doses groups were higher than those in the model group at the same time point (all P<0.05). In the two weeks treatment experiment, the 24 hours urinary cadmium levels of rabbits in medium-dose and high-dose groups at the three time points were higher than those in the model group at the same time point (all P<0.05). In the four weeks treatment experiment, the 24 hours urinary cadmium level on the 19th day of rabbits in the low-dose group was higher than that in the model group at the same time point (P<0.05), and the 24 hours urinary cadmium levels of rabbits in medium- and high-dose groups at the five time points were higher than those in the model group at the same time point (all P<0.05), except for the rabbits of fifth day of the medium-dose group. The kidney cadmium levels of rabbits in the low-dose group after four week of treatment and in the medium- and high-dose groups after one, two, and four weeks of treatment decreased compared with the model group (all P<0.05). No obvious adverse effects were observed during the treatment. ii) The MTD of GMDTC in rats administered intravenously in a single dose was 3 000.0 mg/kg body weight. During the period of intravenous infuseion with GMDTC for four consecutive weeks, the blood drug level reached the peak at the end of the first and last administrations (eight min), and no clinical adverse reactions were observed during this period of time, nor was there any apparent accumulation. The NOAEL for intravenous infusion of GMDTC for four consecutive weeks in rats was 500.0 mg/kg body weight. Conclusion The initial dose of the GMDTC injection in the cadmium poisoning rabbit was 36.0 mg/kg body weight, and the recommended initial dose for human is 480.0 mg/person. Intravenous infusion of GMDTC is characterized by rapid absorption, rapid elimination, and no accumulation.
4.Polymorphisms of host tropism relating amino acid sites in influenza A virus
Xiuliang LIU ; Yanjiao LI ; Weijie CHEN ; Yuxi WANG ; Qile GAO ; Jingjing HU ; Zhijie ZHANG ; Chenglong XIONG
Shanghai Journal of Preventive Medicine 2023;35(7):626-633
ObjectiveTo discover and analyze single or several correlative key amino acid sites that influence the host tropism during the influenza A virus (IAV) infection based on complete internal protein gene segments of IAV strains, and to provide evidence for the study of human host-adaptive mutations of IAV. MethodsThe full-length nucleotide sequences of 43 671 IAV strains containing 6 complete internal gene segments were downloaded from the GISAID EpiFluTM database, and 698 human-tropic (HU) and 1 266 avian-tropic (AV) representative strains were included. The consensus coding sequences of the representative strains from the amphitropic category were compared by R script, and the differential amino acid sites and their polymorphisms were then obtained. The multi-site combination analysis of differential sites was conducted with R script. ResultsA total of 49 and 57 conserved differential sites were obtained from the consensus sequence comparison between AV and H1N1 (subtype from HU), and comparison between AV and H3N2 (another subtype from HU), separately. 79 and 65 multi-site combinations were found between HU and AV strains through 3 and 4 sites combination analysis, respectively, and a total of 11 conserved sites were involved: site 271 and 684 in PB2; site 336, 486, 581 and 621 in PB1; site 204 and 356 in PA; site 33, 305 and 357 in NP. No eligible differential sites were found in M1 and NS1. ConclusionSeveral conserved amino acid differential sites, between HU and AV strains of IAV, are found in PB2, PB1, PA and NP proteins. Instead of working as single units, these sites may have interactions, forming specific amino acid combinations that determine the host tropism of IAV collectively.
5.Clinical study of deformed complex vertebral osteotomy (DCVO) in the treatment of angular kyphosis of cured spinal tuberculosis
Hongqi ZHANG ; Mingxing TANG ; Lige XIAO ; Qile GAO ; Chaofeng GUO ; Shaohua LIU ; Yuxiang WANG ; Ang DENG ; Jinyang LIU
Chinese Journal of Orthopaedics 2021;41(12):744-754
Objective:To evaluate the feasibility and clinical efficacy of deformed complex vertebral osteotomy (DCVO) technique on the treatment of angular kyphosis of cured spinal tuberculosis.Methods:A retrospective study was performed on patients with angular kyphosis of cured spinal tuberculosis who underwent the DCVO technique or posterior vertebral column resection (PVCR) technique from Jan, 2007 to Jan, 2019. 33 patients were included, 18 males and 15 females, the average age was 39.5±15.0 years old (ranged 9-78 years old). The vertebral deformity in thoracic vertebrae 14 cases, thoracolumbar vertebrae 16 cases, and lumbar vertebrae 3 cases. 20 cases underwent the DCVO technique, while 13 cases underwent PVCR technique. For DCVO group, the multiple malformed vertebrae were considered a malformed complex, and a larger range and angle wedge osteotomy was performed within the complex using the DCVO technique. PVCR technique would resect the whole deformed vertebrae, and subsequently brought the two separated spinal columns together with instruments and titanium mesh. The intro-operative blood loss, operating time and complications were recorded. The radiological measurements included preoperative and postoperative spinopelvic parameters, which including thoracic kyphosis (TK), lumbar lordosis (LL), sagittal vertical axis (SVA), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS) and segmental kyphosis. The recovery of neurological function was evaluated by Frankle classification.Results:All patients were followed up for 7-72 months. Comparing with the cases underwent PVCR technique, the DCVO group has a significantly lower blood loss (1315.00±462.57 ml), operating time (293.00±83.86 min) and complications rate (1.5%). At the time of preoperation, postoperation and last follow-up, the deformity angle of DCVO group was 96.80°±6.32°, 29.10°±6.96°and 29.05°±6.49°, which gained an average 69.9% correction rate. The statistical analysis suggested that deformity angle was enormously corrected. And there was an insignificant difference between DCVO group and PVCR group. Meanwhile, the preoperative, postoperative and follow-up TK of DCVO group was 96.96°±29.13°, 37.15°±4.88° and 37.00°±3.89°respectively, whosecorrection rate was 67.1%; LL was 66.70°±21.21°, 42.25°±5.53° and 41.90°±4.98°, which have a significant difference between pre-operation and post-operation/follow-up ( F=23.997, P<0.001) ; SVA was 75.95±18.63 mm, 16.30±6.88 mm and 16.55±7.30 mm. PI was 47.50°±6.12°, 47.35°±5.54°and 47.90°±5.93°, PT was 37.25°±9.63°, 18.50°±1.99° and 19.00°±1.65°; SS was 10.25°±8.27°, 29.15°±5.91° and 28.85°±5.77°. The sagittal and spinopelvic parameters of two groups improved significantly at postoperation and follow-up. No obviously difference of spinal parameters was found between two groups at preoperation and postoperation. Both groups have cases with dysneuria. And all of these cases achieved different degrees of recovery at follow-up. Conclusion:The use of DCVO technique for the treatment of post-tubercular angular kyphosis is safe and efficiency. DCVO leads a better clinical outcomes and lower complication rate than VCR technique.
6.Refractive development during the past 5 years among primary school students in Jinshan District of Shanghai
GAO Qile, ZHOU Xiaodong, QI Huihong, CHEN Xuefeng, LI Tao
Chinese Journal of School Health 2019;40(2):268-269
Objective:
To explore refractive development of primary school students in Jinshan district of Shanghai during the past five years.
Methods:
A total of 201 first grade pupils enrolled in 2013 were followed up for 5 years. Annual examination of non-cycloplegic refraction and axial length (AL) was implemented and analyzed.
Results:
The mean spherical equivalents (SEs) of boys at each grade was (0.22±0.53)(-0.04±0.64)(-0.36±0.92)(-0.74±1.23)(-1.14±1.67)D, respectively; for girls, (0.26±0.88)(-0.03±1.02)(-0.28±1.02)(-0.64±1.32)(-1.13±1.65)D, respectively. The mean ALs of boys at each grade was (22.94±0.60)(23.13±0.68)(23.45±0.69)(23.65±0.81)(24.03±0.93)mm, respectively and was (22.40±0.67)(22.67±0.70)(22.95±0.74)(23.14±0.79)(23.59±0.90)mm for girls at each grade, respectively. There were negative correlations between dioptres and ALs in each grade(r=-0.26, -0.35, -0.41, -0.53, -0.59, P<0.05).
Conclusion
The dioptre and AL among primary school students in Jinshan district of Shanghai increased gradually and results in developing into myopia. The dioptre negatively associates with AL, which should be both paid attention to among primary school students.
7.Expanding PSO technique for the treatment of congenital severe thoracic angular kyphotic deformity
Hongqi ZHANG ; Lige XIAO ; Chaofeng GUO ; Mingxing TANG ; Jinyang LIU ; Jianghuang WU ; Ang DENG ; Yuxiang WANG ; Qile GAO
Chinese Journal of Orthopaedics 2017;37(22):1377-1384
Objective To evaluate the clinical efficacy and feasibility of using the expanding pedicle subtraction osteotomy (E-PSO) technique for the treatment of congenital severe thoracic angular kyphotic deformity.Methods We retrospectively reviewed a cohort of 13patients with congenital severe kyphosis admitted to our hospital from January 2010 to June 2015 including 5 males and 8 females,the average age is (34.9±20.5) years old (ranged 15-55 years old).The vertebral deformity in T7~83 cases,T8~93 cases,T9~102 cases,T10~114 cases,T9~111 case.All cases were treated by E-PSO technique.The multi-malformed vertebrae are considered as a complexus.And the osteotomy was performed within the complexus.The superior and inferior endplate of the complexus were reserved.After the osteotomy was completed,alternately pressed tightly closed the upper and lower parts.Results All cases were followed up for 10-42 months,with an average of 32 months.At the time of preoperation,postoperation andthe last follow-up,the deformity angle was 107.0°±3.5°,23.5°± 1.5° and 23.5°±0.2°;TK was 98.1°±7.6°,28.9°±3.0° and 29.5°±0.1 °,LL was 94.1 °± 1.5°,43.7°± 1.3° and 44.1 °±5.3°;SVA was (-0.6±39) mm,(1.6±7.9) mm and (6±0.7) mm,respectively;PI was 28.9°±1.6°,31.7±12.3°and 31.9°±2.1°;PT was 17.7°±1.9°,13.4°±3.4°and 13.1°±4.2°,SS was 11.3°±0.4°,18.2°±1.1° and 18.7°±2.1°,respectively.The sagittal parameters and spinopelvic parameters except SVA were significantly improved in the post-operation and the last follow-up compared with the pre-operation according to the image data.No significant loss of correction occurred during the follow-up,and there was no statistical difference.The preoperative VAS score was (5.7± 1.4) points,ODI score was (19.8±12.7) points.The last follow-up VAS score was (1.9±0.7) points,the ODI score was (9.2±0.7) points.No case of nerve damage,infection and other complications,and no dissection,displacement and rupture of internal fixation were found during the follow-up.Conclusion The use of E-PSO technique for the treatment of congenital severe thoracic kyphotic kyphosis is feasible and can achieve better curative effect.
8.Posterior only approaches versus anterior only approaches and combined posterior and anterior approaches for lumbar tuberculosis in adults:minimum 5-year follow-up
Hongqi ZHANG ; Qiang GUO ; Chaofeng GUO ; Jianhuang WU ; Jinyang LIU ; Qile GAO ; Yuxiang WANG ; Xiyang WANG
Chinese Journal of Orthopaedics 2016;36(11):651-661
Objective To compare the minimum 5?year follow?up outcomes of surgical management by posterior only, anterior only and combined posterior and anterior approaches for lumbar tuberculosis in adults, evaluate the mid?term follow?up results of the surgery for the treatment of lumbar tuberculosis and explore its advantages and indications. Methods From Jun 2004 to Jan 2010, 311 adult patients with lumbar tuberculosis were treated surgically. The clinical data of 137 cases that met the enrolled criteria and had integrity following?up data was analyzed retrospectively. It included the patients who had the surgical indication of the posterior only surgery but underwent the anterior only or the combined posterior and anterior ap?proaches before 2008. There were 83 cases of male and 54 cases of female. The age ranged from 20 to 75 years, with a mean of 65.6 years. Among these patients, 63 cases were treated with single?stage posterior debridement, interbody fusion and instru?mentation (the posterior group); 42 cases were treated with posterior instrumentation, and anterior debridement and bone graft in a single or two?stage procedures (the combined group) and 32 cases were treated with anterior debridement and strut graft?ing with instrumentation (the anterior group). Trauma index (the operation time, blood loss, the length of hospital stay, compli?cations);imaging parameters (Segment kyphotic angle, corrective rate, loss angle, bone fusion time) and the quality?of?life indi?cators (Oswestry Disability Index、Frankle grade、visual analogue scale、Macnab score) were compared among three groups. Re?sults The mean operation time, mean blood loss and the complications rate were (207.9 ± 30.9) min, (409.5 ± 107.9) ml and 12.95%in the posterior group;(270.7±32.0) min, (649.0±120.0) ml and 30.95%in the anterior group;(349.7±38.9) min, (840.0± 168.7) ml and 25%in the combined group. The operation time, blood loss and the complications rate of the posterior group were less than the anterior group and the combined group, and the difference was significant;The combined group consumed the longest operation time, associated with the most intraoperative blood loss, the highest complication rate and the longest hospital stay among the three groups, and the difference was significant. The correction rate of kyphosis achieved of the anterior group ( 52%± 5.45%) was significantly inferior to the posterior group (74%±5.04%) and the combined group (69%±7.95%), while the loss of cor?rection in the anterior group (2.5°) was higher than both the posterior group (0.8°) and the combined group (1.1°), and the differ?ence was significant. The average follow?up was(6.5±1.96)years (range, 5-11). The mean bone fusion time of the posterior group, the anterior group and the combined group were (6.0±1.5) months, (6.2±1.3) months and (6.5±1.6) months respectively, and there was no statistic difference. After the surgery, the quality of life was improved obviously in all patients. At the time of the latest fol?low?up, the improvement rate of the ODI,VAS and the excellent and good rate according to the Macnab score were 80.6%±2.1%, 81.7%± 1.6%and 95.24%in the posterior group;79.8%± 1.5%, 79.7%± 2.0%and 92.95%in the anterior group;81.3%± 1.1%, 79.9%±0.8%and 90.63%in the combined group. There was no significant difference among the groups in the improvement rates of the ODI, VAS, Frankel grade and the excellent and good rate of the Macnab score. Conclusion The Mid?term follow?up of the different surgical procedures for the treatment of the lumbar tuberculosis in adults were basically satisfactory. Compared with the traditional surgery, the posterior?only surgery is a safe, minimally invasive and effective method in the management of monoseg?ment lumbar tuberculosis in adults.
9.Posterior only approaches versus anterior only approaches and combined posterior and anterior approaches for thorac-ic tuberculosis in adults:minimum 5-year follow-up
Hongqi ZHANG ; Longjie WANG ; Mingxing TANG ; Qile GAO ; Jinyang LIU ; Jianhuang WU ; Jianzhong HU
Chinese Journal of Orthopaedics 2016;36(11):641-650
Objective To compare the minimum 5?year follow?up outcomes of surgical management by posterior only ap?proaches, anterior only approaches and combined posterior and anterior approaches for thoracic tuberculosis in adults, and evalu?ate the mid term follow?up results of posterior only approaches. Methods All of 184 patients with monosegment thoracic tubercu?losis between January 2003 and November 2010 were studied retrospectively. Among these patients, 62 cases were treated with posterior debridement combine with interbody fusion (PO group), 65 cases were treated by posterior instrumentation, anterior de?bridement and bone graft in one or two?stage procedures (AP Group ), and 57 cases were treated by anterior only approach (AO Group). The operation time, blood loss, Visual Analogue Scale, complications, recovery of neurological function, kyphosis angle, correction rate and loss angle were respectively compared between each group. Results Comparison of postoperative curative ef?fects showed:mean operation time and blood loss:PO group (260.05±30.75 min,735.95±161.43 ml) was better than AP group (411.65 ± 55.61 min, 1178.65 ± 184.50 ml)and AO group (343.65 ± 24.74 min, 965.35 ± 122.59 ml);corrective angle and correction rate:PO group (6.78°±1.13°, 72.48%±12.97%) and AP group (6.97°±1.05°, 73.10%±11.42%) were better than AO group (13.98°± 1.73°, 44.95%±16.84%);bed time:PO group and AO group were shorter than AP group. Mid term follow?up outcomes showed:ky?phosis angle and loss angle:PO group (8.56°±1.09°, 1.89°±1.41°) and AP group (8.55°±1.65°, 1.63°±1.11°) were better than AO group (16.39°±1.59°, 2.80°±1.29°);bone fusion time, VAS and recovery of neurological function:there were no statistically differ?ence in all groups. Conclusion The mid term follow?up outcomes of posterior debridement combined with interbody fusion is sat?isfied in the management of monosegment thoracic tuberculosis. It is a safe and effective method.
10.Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts
Ang DENG ; Hongqi ZHANG ; Chaofeng GUO ; Yuxiang WANG ; Qile GAO ; Mingxing TANG ; Shaohua LIU ; Jinyang LIU
Chinese Journal of Tissue Engineering Research 2016;20(29):4261-4268
BACKGROUND:There are few studies concerning estrogen receptorβgene, and its mechanism of regulating the bone metabolism is stil unclear now. OBJECTIVE:To analyze the effect of estrogen receptorβ(ERβ) silencing on the expressions of transforming growth factorβ1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS:There were three groups:blank control group (hFOB 1.19 uninfected with any retrovirus);negative control group (containing invalid interference fragment ERβ-shRNA-nc);optimal RNAi group (ERβ-shRNA-3). ERβ-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts fol owed by resistance screening and cel expansion. MTT assay was used to detect the proliferative activity of ERβ-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ERβwas determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ERβsilencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION:Human osteoblasts that were stably transfected by ERβ-shRNA-3 retroviral vector was selected successful y, and ERβsilencing had no significant influence on the cel proliferation (P>0.05). Under the interference of estrogen, the silencing efficiency of ERβprotein was (93.11±0.57)%(P<0.05), and after ERβsilencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)%and (16.62±1.71)%at mRNA level, and increased by (23.79±3.76)%and (18.08±3.20)%at protein level (both P<0.05). In conclusion, ERβmay play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2.


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