A multiprotein signaling complex sustains AKT and mTOR/S6K activity necessary for the survival of cancer cells undergoing stress.

Cancer cells encounter stresses during tumor progression and metastatic spread, however, how they survive these challenges is not fully understood. We now identify a mechanism for cancer cell survival through the discovery of a multiprotein signaling complex that includes the GTPase Cdc42, the Cdc42 GEF/effector protein Dock7, AKT, mTOR and the mTORC1 regulatory partners TSC1, TSC2, and Rheb. This pro-survival signaling complex sustains the activated state of AKT by preventing its dephosphorylation at Ser473 during serum starvation, resulting in a low but critical activation of a Raptor-independent mTOR/S6K activity. We demonstrate that the Dock7 DHR1 domain, previously of unknown function, is responsible for preserving AKT phosphorylation through an interaction requiring its C2-like motif. Collectively, these findings help address long-standing questions of how Cdc42 signals mTOR activation by elucidating the unique functions of its signaling partner Dock7 as an AKT regulator necessary for resistance to anoikis and apoptosis in cancer cells.

Inhibiting Glutamine Metabolism Blocks Coronavirus Replication in Mammalian Cells.

Developing therapeutic strategies against COVID-19 has gained widespread interest given the likelihood that new viral variants will continue to emerge. Here we describe one potential therapeutic strategy which involves targeting members of the glutaminase family of mitochondrial metabolic enzymes (GLS and GLS2), which catalyze the first step in glutamine metabolism, the hydrolysis of glutamine to glutamate. We show three examples where GLS expression increases during coronavirus infection of host cells, and another in which GLS2 is upregulated. The viruses hijack the metabolic machinery responsible for glutamine metabolism to generate the building blocks for biosynthetic processes and satisfy the bioenergetic requirements demanded by the 'glutamine addiction' of virus-infected host cells. We demonstrate how genetic silencing of glutaminase enzymes reduces coronavirus infection and that newer members of two classes of small molecule allosteric inhibitors targeting these enzymes, designated as SU1, a pan-GLS/GLS2 inhibitor, and UP4, which is specific for GLS, block viral replication in mammalian epithelial cells. Overall, these findings highlight the importance of glutamine metabolism for coronavirus replication in human cells and show that glutaminase inhibitors can block coronavirus infection and thereby may represent a novel class of anti-viral drug candidates. Teaser: Inhibitors targeting glutaminase enzymes block coronavirus replication and may represent a new class of anti-viral drugs.

Apophysomyces variabilis as an emerging pathogen in Mainland China.

BACKGROUND: Mucormycosis is a rare form of invasive, rapidly progressive and lethal opportunistic fungal infection caused by Mucorales. Although Rhizopus arrhizus (R. arrhizus) is the most commonly isolated Mucorales worldwide, infections caused by Apophysomyces variabilis (A. variabilis) are increasing. OBJECTIVES AND METHODS: We present a case of necrotizing fasciitis caused by A. variabilis in an immunocompetent woman. In order to further understand the characteristics of the strain isolated from the patient, we identified the strain through ITS sequencing, assessed the ability to tolerate salt concentrations and temperature conditions, in addition to performing in vitro drug susceptibility testing against common antifungal agents. RESULTS: The strain showed 98.76% identity with A. variabilis in the NCBI database, and it was found to tolerate higher temperatures and salt concentrations than previously reported strains. The strain was sensitive to amphotericin B and posaconazole, but not to voriconazole, itraconazole, 5-fluorocytosine and echinocandins. CONCLUSIONS: This case indicates that Mucorales caused by A. variabilis should be recognised as an emerging pathogen that can cause a high mortality rate in the absence of prompt diagnosis and proper treatment in China, aggressive surgical debridement combined with prompt and appropriate antifungal treatment may improve outcomes.

Posterior long segment instrumentation combined with transpedicular impaction bone grafting or with bone cement augmentation in the treatment of stage III Kümmell disease: a comparison of efficacies / 中华创伤杂志

Objective:To compare the efficacies of posterior long segment instrumentation combined with transpedicular impaction bone grafting or with bone cement augmentation in treating stage III Kümmell disease.Methods:A retrospective cohort study was conducted to analyze the clinical data of 38 patients with stage III Kümmell disease who were admitted to Zhengzhou Orthopedic Hospital between January 2016 and December 2020. The study included 8 male and 30 female patients, with ages ranging from 59 to 81 years [(68.9±4.9)years]. The vertebral fractures occurred at T 8 in 1 patient, T 11 in 9 patients, T 12 in 10 patients, and L 2 in 10 patients. Seventeen patients underwent posterior long segment instrumentation combined with transpedicular impaction bone grafting (impaction bone grafting group), and 21 patients underwent posterior long segment instrumentation combined with bone cement augmentation (bone cement group). The surgical duration, intraoperative blood loss, and incidences of postoperative complications were compared between the two groups. Additionally, the visual analogue score (VAS), Japanese orthopedic association (JOA) score, and Cobb angle were compared before the operation, at 1 week and 3 months post-operation, and at the final follow-up for both groups. The study also compared bone healing at the last follow-up and postoperative complication rates between the two groups. Results:All the patients were followed up for 24-35 months [(28.7±2.9)months]. The impaction bone grafting group had a surgical duration of (150.7±25.4)minutes and intraoperative blood loss of (285.3±48.6)ml, significantly different from those in the bone cement group [(132.0±21.1)minutes, (251.4±44.8)ml] (all P<0.05). Before the operation, there were no significant differences in the VAS, JOA score, or Cobb angle between the two groups (all P>0.05).The VAS was (3.2±0.8)points, (2.7±0.5)points and (2.2±0.7)points in the impaction bone grafting group and was (2.7±0.6)points, (2.6±0.7)points and (2.4±0.8)points in the bone cement group at 1 week and 3 months post-operation and at the final follow-up, respectively. The VAS in the impaction bone grafting group was significantly higher than that in the bone cement group at 1 week post-operation ( P<0.05); however, no significant differences were found at 3 months post-operation or at the last follow-up (all P>0.05). There was no significant difference in the JOA score between the two groups at 1 week or 3 months post-operation, or at the final follow-up (all P>0.05). The Cobb angle in the impaction bone grafting group was (5.1±1.3)°, (5.9±1.8)° and (6.5±2.5)° at 1 week and 3 months post-operation, and at the final follow-up, significantly lower than that in the bone cement group [(8.4±1.6)°, (12.6±2.1)°, and (14.5±3.3)°] (all P<0.01). All the patients in the impaction bone grafting group achieved bone healing at the last follow-up. One patient in the impaction bone grafting group experienced delayed incision healing, whereas two patients in the bone cement group had poor bone healing. The complication rate was 5.9% (1/17) in the impaction bone grafting group and 9.5% (2/21) in the bone cement group ( P>0.05). Conclusions:Posterior long segment instrumentation combined with transpedicular impaction bone grafting or with bone cement augmentation are both effective in alleviating pain and improving the spinal function for stage III Kümmell disease. The former procedure is associated with longer surgical duration and increased intraoperative blood loss, but it can provide superior correction and maintenance of kyphosis deformity, promoting the healing of the injured vertebrae.

Precision targeted and traditional percutaneous vertebroplasty in treating refracture of injured vertebra after operation for Kümmell disease: a comparison of efficacies / 中华创伤杂志

Objective:To compare the clinical efficacies of precision targeted and traditional percutaneous vertebroplasty (PVP) in the treatment of refracture of injured vertebra after operation for Kümmell disease.Methods:A retrospective cohort study was conducted to analyze the clinical data of 23 Kümmell disease patients suffering from refracture of injured vertebra after PVP in Zhengzhou Orthopedic Hospital from October 2014 to October 2018. The patients included 7 males and 16 females, aged 53-89 years [(69.3±3.5)years]. There were 11 patients of stage I Kümmell disease and 12 patients of stage II Kümmell disease. The vertebral distribution of fracture was T 11 (3 patients), T 12 (9 patients), L 1 (8 patients) and L 2 (3 patients). Eleven patients received traditional PVP treatment (traditional PVP group) and 12 patients received precision targeted PVP treatment (targeted PVP group). The operation time, amount of bone cement injection and filling of bone cement in the fracture space were compared between the two groups. The visual analogue score (VAS) and Oswestry disability index (ODI) were also compared before operation, at 2 days, 1 month, 3 months, 6 months after operation, and at the last follow-up. The rates of bone cement leakage and re-collapse of injured vertebra were observed in the two groups. Results:The patients were followed up for 12-36 months [(24.2±2.6)months]. There were no significant differences in the operation time or amount of bone cement injection between the two groups (all P>0.05). All the fracture spaces in the targeted PVP group were fully filled with bone cement, while 4 patients in the traditional PVP group showed inadequate filling of the fracture area ( P<0.05). The VAS values in the targeted PVP group were (8.9±0.5)points, (1.6±0.2)points, (1.7±0.1)points, (1.8±0.1)points, (1.9±0.3)points, and (1.8±0.4)points before operation, at 2 days, 1 month, 3 months, 6 months after operation and at the last follow-up; and those in the traditional PVP group were (9.1±0.9)points, (1.8±0.4)points, (1.8±0.2)points, (2.0±0.4)points, (2.1±0.2)points, and (2.4±0.3)points, respectively. The VAS values of both groups were significantly decreased at 2 days, 1 month, 3 months, 6 months after operation, and at the last follow-up compared with those before operation (all P<0.05), but there was no significant difference between different time points after operation (all P>0.05). No significant differences were found in the VAS values between the two groups before operation and at 2 days, 1 month, 3 months and 6 months after operation (all P>0.05). However, the VAS value in the targeted PVP group was significantly lower than that in the traditional PVP group at the last follow-up ( P<0.05). The ODI values in the targeted PVP group were 38.5±4.3, 7.2±2.3, 7.3±2.0, 7.2±1.8, 7.3±2.4, and 7.4±2.5 before operation and at 2 days, 1 month, 3 months, 6 months after operation, and at last follow-up; and those in the traditional PVP group were 37.8±4.1, 7.5±2.5, 7.7±1.9, 7.9±2.4, 8.1±2.6, and 9.6±2.4, respectively. The ODI values of both groups were significantly decreased at 2 days, 1 month, 3 months, 6 months after operation and at the last follow-up compared with those before operation (all P<0.05), but there were no significant differences between different time points after operation (all P>0.05). The ODI values were not significantly different between the two groups before operation and at 2 days, 1 month, 3 months, 6 months after operation (all P>0.05), but the ODI value in the targeted PVP group was significantly lower than that in the traditional PVP group at the last follow-up ( P<0.05). There were no significant differences in the rates of bone cement leakage or re-collapse of injured vertebra between the two groups (all P>0.05). Conclusion:Compared with traditional PVP treatment for refracture of injured vertebra after operation for Kümmell disease, targeted PVP can make bone cement injection fully dispersed, greatly reduce pain and promote functional recovery.

Classified reduction based on CT two-dimensional images for the surgical treatment of single segment facet joint dislocation in subaxial cervical spine / 中华创伤杂志

Objective:To investigate the efficacy of the classified reduction based on CT two-dimensional images for the surgical treatment of single segment facet joint dislocation in subaxial cervical spine.Methods:A retrospective case series study was made on 105 patients with single segment facet joint dislocation in subaxial cervical spine admitted to Zhengzhou Orthopedic Hospital from January 2015 to October 2022. There were 63 males and 42 females, with the age range of 22-78 years [(47.5±3.6)years]. Preoperative American Spinal Cord Injury Association (ASIA) classification was grade A in 23 patients, grade B in 45, grade C in 22, grade D in 15 and grade E in 0. The classification of surgical approach was based on the presence or not of continuity between anterior and posterior subaxial cervical structures and the movability of the posterior cervical facet joint on CT two-dimensional images, including anterior cervical surgery if both were presented and posterior facet joint resection plus anterior cervical surgery if there was discontinuity between anterior and posterior subaxial cervical structures or posterior facet joint fusion. Reduction procedures were applied in accordance with the type of facet joint dislocation classified based on the position of the lower upper corner of facet joint, including skull traction or manipulative reduction for the dislocation locating at the dorsal side (type A), intraoperative skull traction and leverage technique for the dislocation locating at the top (type B) and intraoperative skull traction and leverage technique with boosting for the dislocation locating at the ventral side (type C). If the dislocation of two facet joints in the same patient was different, the priority of management followed the order of type C, type B and type A. The reduction success rate, operation time and intraoperative blood loss were recorded. The cervical physiological curvature was evaluated by comparing the intervertebral space height and Cobb angle before operation, at 3 months after operation and at the last follow-up. The fusion rate of intervertebral bone grafting was evaluated by Lenke grading at 3 months after operation. The spinal cord nerve injury was assessed with ASIA classification before operation and at 3 months after operation. Japanese Orthopedic Association (JOA) score was applied to measure the degree of cervical spinal cord dysfunction before operation and at 3 months after operation, and the final follow-up score was used to calculate the rate of spinal cord functional recovery. The occurrence of complications was observed.Results:All patients were followed up for 3-9 months [(6.0±2.5)months]. The reduction success rate was 100%. The operation time was 40-95 minutes [(58.6±9.3)minutes]. The intraoperative blood loss was 40 to 120 ml [(55.7±6.8)ml]. The intervertebral space height was (4.7±0.3)mm and (4.7±0.2)mm at 3 months after operation and at the last follow-up, significantly decreased from preoperative (3.1±0.5)mm (all P<0.01), but there was no significant difference in intervertebral space height at 3 months after operation and at the last follow-up ( P>0.05). The Cobb angle was (6.5±1.3)° and (6.3±1.2)° at 3 months after operation and at the last follow-up, significantly increased from preoperative (-5.4±2.2)° (all P<0.01), but there was no significant difference in Cobb angle at 3 months after operation and at the last follow-up ( P>0.05). The fusion rate of intervertebral bone grafting evaluated by Lenke grading was 100% at 3 months after operation. The ASIA grading was grade A in 15 patients, grade B in 42, grade C in 29, grade D in 12 and grade E in 7 at 3 months after operation. The patients showed varying degrees of improvement in postoperative ASIA grade except that 15 patients with preoperative ASIA grade A had partial recovery of limb sensation but no improvement in ASIA grade. The JOA score was (13.3±0.6)points and (13.1±0.6)points at 3 months after operation and at the last follow-up, significantly improved from preoperative (6.8±1.4)points (all P<0.01), but there was no significant difference in JOA score at 3 months after operation and at the last follow-up ( P>0.05). The rate of spinal cord functional recovery was (66.3±2.5)% at the last follow-up. All patients had no complications such as increased nerve damage or vascular damage. Conclusion:The classified reduction based on CT two-dimensional images for the surgical treatment of single segment facet joint dislocation in subaxial cervical spine has advantages of reduced facet joint dislocation, recovered intervertebral space height and physiological curvature, good intervertebral fusion and improved spinal cord function.

Study on the mechanisms of action of berberine combined with fluconazole against fluconazole-resistant strains of Talaromyces marneffei.

Talaromyces (Penicillium) marneffei (T. marneffei) is a thermally dimorphic fungus that can cause opportunistic systemic mycoses. Our previous study demonstrated that concomitant use of berberine (BBR) and fluconazole (FLC) showed a synergistic action against FLC-resistant T. marneffei (B4) in vitro. In this paper, we tried to figure out the antifungal mechanisms of BBR and FLC in T. marneffei FLC-resistant. In the microdilution test, the minimum inhibitory concentration (MIC) of FLC was 256 µg/ml before FLC and BBR combination, and was 8 µg/ml after combination, the partial inhibitory concentration index (FICI) of B4 was 0.28. After the treatments of BBR and FLC, the studies revealed that (i) increase reactive oxygen species (ROS), (ii) reduce ergosterol content, (iii) destroy the integrity of cell wall and membrane, (iv) decrease the expression of genes AtrF, MDR1, PMFCZ, and Cyp51B however ABC1 and MFS change are not obvious. These results confirmed that BBR has antifungal effect on T. marneffei, and the combination with FLC can restore the susceptibility of FLC-resistant strains to FLC, and the reduction of ergosterol content and the down-regulation of gene expression of AtrF, Mdr1, PMFCZ, and Cyp51B are the mechanisms of the antifungal effect after the combination, which provides a theoretical basis for the application of BBR in the treatment of Talaromycosis and opens up new ideas for treatment of Talaromycosis.

Experimental Phaeohyphomycosis of Curvularia lunata.

Originally considered to be a plant pathogen, reports of phaeohyphomycosis due to Curvularia lunata (C. lunata) in animals and humans are increasing. However, studies on the pathogenesis, virulence, and epidemiology of C. lunata have rarely been discussed. In the present study, BALB/c mice were experimentally inoculated with C. lunata suspension by different routes and the course of infection was evaluated. In addition, the in vitro antifungal susceptibility of C. lunata against six commonly used antifungals was evaluated using the microdilution method. Inoculation resulted in skin lesions in animals inoculated intraperitonially and subcutaneously. Infection was confirmed by both mycological and histopathologic examination. C. lunata spores and hyphae were detected in the histopathologic sections stained with hexamine silver staining. In addition, voriconazole (VRC) demonstrated greater activity against C. lunata when compared to the other antifungals, whereas fluconazole (FLC) was the least active antifungal with a minimum inhibitory concentration (MIC) range of 8-16 µg/mL. Further studies are necessary to understand the pathogenicity of C. lunata and uncover the mystery of this fungus.

Transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow in the treatment of stage III Kümmell′s disease / 中华创伤杂志

Objective:To compare the clinical effect of transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation and posterior subtotal vertebrectomy combined with posterior internal fixation in the treatment of stage III Kümmell′s disease.Methods:A retrospective cohort study was made on clinical data of 40 patients with stage III Kümmell′s disease admitted to Zhengzhou Orthopedic Hospital from June 2015 to December 2018. There were 10 males and 30 females, at age range of 57-79 years［(67.7±6.1)years］. A total of 19 patients were treated by transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation (impaction bone graft group), and 21 patients by posterior subtotal vertebrectomy combined with posterior internal fixation (subtotal vertebrectomy group). Operation time and intraoperative blood loss were compared between the two groups. Degree of pain, lumbar dysfunction and degree of kyphosis were evaluated by visual analogue scale (VAS), Japanese Orthopedic Association (JOA) score and kyphotic Cobb angle before operation, at 1 week after operation and at the last follow-up. Bone healing time was compared between the two groups. The complications of the two groups were observed.Results:All patients were followed up for 25-64 months［(40.6±10.4)months］. Operation time and intraoperative blood loss were (130.0±10.1)minutes and (284.5±43.5)ml in impaction bone graft group, lower than those in subtotal vertebrectomy group［(253.8±33.2)minutes, (889.1±95.7)ml］(both P<0.01). There were no significant differences in VAS, JOA score or kyphotic Cobb angle between the two groups before operation, at 1 week after operation and at the last follow-up (all P>0.05). Both VAS and JOA score showed significant differences within each group at any time point (all P<0.01). In both groups, the kyphotic Cobb angle reduced significantly at 1 week after operation when compared with that before operation (all P<0.01), and the angle showed a slight increase at the last follow-up, but remained significantly lower than that before operation (all P<0.01). There were no relapse of pain or aggravation of kyphosis. Bone healing time in impaction bone graft group［4.4(4.0, 5.0)months］was significantly shorter than that in subtotal vertebrectomy group［6.4(5.2, 8.1)months］( P<0.01). There were 2 patients with delayed healing of surgical incision in impaction bone graft group, with the complication rate of 11%. There were 2 patients with dural tear and 3 patients with delayed healing of surgical incision in subtotal vertebrectomy group, with the complication rate of 24%. The complication rate was not statistically significant between the two groups ( P>0.05). No loosening or breakage of internal fixation was observed during the follow-up. Conclusions:Transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation and posterior subtotal vertebrectomy combined with posterior internal fixation are effective in the treatment of stage III Kümmell disease. However, the former can shorten the operation time, reduce the intraoperative blood loss and accelerate the healing of injured vertebral bone, suggesting a relatively minimally invasive surgical method for reconstruction and maintenance of spinal biomechanical stability.

In vitro Antifungal Susceptibility Profiles of Cryptococcus neoformans var. grubii and Cryptococcus gattii Clinical Isolates in Guangxi, Southern China.

This study analyzed the in vitro drug sensitivity of Cryptococcus spp. from Guangxi, Southern China. One hundred three strains of Cryptococcus were recovered from 86 patients; 14 were HIV positive and 72 were HIV negative. Ninety-two strains were identified as Cryptococcus neoformans var. grubii, while 11 strains were identified as Cryptococcus gattii (5 C. gattii sensu stricto and 6 Cryptococcus deuterogattii). The recovered strains were tested against commonly used antifungal drugs (fluconazole, amphotericin B, 5-fluorocytosine, itraconazole, and voriconazole) and to novel antifungal drugs (posaconazole and isavuconazole) using CLSI M27-A4 method. The results showed that all isolates were susceptible to most antifungal drugs, of which the minimum inhibitory concentration (MIC) ranges were as follows: 0.05-4 µg/ml for fluconazole, 0.25-1 µg/ml for amphotericin B; 0.0625-2 µg/ml for 5-fluorocytosine, 0.0625-0.25 µg/ml for itraconazole, 0.0078-0.25 µg/ml for voriconazole, 0.0313-0.5 µg/ml for posaconazole, 0.0020-0.125 µg/ml for isavuconazole for C. neoformans var. grubii isolates, and 1-16 µg/ml for fluconazole, 0.125-1 µg/ml for 5-fluorocytosine, 0.25-1 µg/ml for amphotericin B, 0.0625-0.25 µg/ml for itraconazole, 0.0156-0.125 µg/ml for voriconazole, 0.0156-0.25 µg/ml for posaconazole, and 0.0078-0.125 µg/ml for isavuconazole for C. gattii isolates. Furthermore, some C. neoformans var. grubii isolates were found to be susceptible-dose dependent to 5-fluorocytosine and itraconazole. In addition, a reduction in the potency of fluconazole against C. gattii is possible. We observed no statistical differences in susceptibility of C. neoformans var. grubii and C. gattii in the tested strains. Continuous observation of antifungal susceptibility of Cryptococcus isolates is recommended to monitor the emergence of resistant strains.

Cdc42 functions as a regulatory node for tumour-derived microvesicle biogenesis.

Tumour-derived microvesicles (MVs) serve as critical mediators of cell-to-cell communication in the tumour microenvironment. So far, the underlying mechanisms of MV biogenesis, especially how key tumorigenesis signals such as abnormal EGF signalling regulates MV release, remain unclear. Here, we set out to establish reliable readouts for MV biogenesis and then explore the molecular mechanisms that regulate MV generation. We found that Rho family small G protein Cdc42 is a convergent node of multiple regulatory signals that occur in MV biogenesis. The binding of activated GTP-bound Cdc42 and its downstream effector, Ras GTPase-activating-like protein 1 (IQGAP1), is required for MV shedding. Activated Cdc42 maintains sustained EGF signalling by inhibiting the internalization of cell surface receptors, including EGFR and the VEGF oligomer, VEGF90K, and then facilitates MV release. Subsequently, we further demonstrated that blocking these signalling pathways using the corresponding mutants effectively reduced MV shedding and significantly inhibited MV-promoted in vivo tumour angiogenesis. These findings reveal a complex regulation of MV shedding by tumour cells, shedding light on the regulatory mechanism of MV biogenesis, and potentially contributing to strategies that target MVs in cancer therapy.

Stage I anterior cervical reduction, decompression, interbody fusion and internal fixation of single-segment lower cervical injury of AO type C subtype F4 / 中华创伤杂志

Objective:To investigate the efficacy of stage I anterior cervical reduction, decompression, interbody fusion and internal fixation of single-segment lower cervical injury of AO type C subtype F4.Methods:A retrospective case series study was made on 45 patients with single-segment lower cervical injury of AO C type F4 subtype admitted to Zhengzhou Orthopedic Hospital from January 2012 to December 2019. The study included 31 males and 14 females with the age of (48.5±3.7)years (range, 23-78 years). Segment of injury was located at C 4/5 in 11 patients, at C 5/6 in 19, and at C 6/7 in 15. Under general anesthesia, all patients (16 unilateral injury and 29 bilateral injury) underwent stage I anterior cervical reduction, decompression, interbody fusion and internal fixation within 24 hours after injury. The interbody fusion methods included autogenous iliac bone in 28 patients and cervical fusion cage in 17. The operation time, blood loss, reduction time and correction rate of zygapophysial joints, postoperative complications and incision healing were analyzed. The height of cervical intervertebral space and Cobb angle were measured through X-ray of lateral cervical vertebrae before operation and3 months after operation to assess the cervical physiological curvature. The type of injury was clarified according to Lenke classification through coronal and sagittal CT scanning to determine the intervertebral bone graft fusion rate. The intraspinal spinal cord decompression was observed through MRI. The nerve function was assessed before operation and 3 months after operation using American Spinal Injury Association (ASIA) scale and Japanese Orthopedic Association (JOA) score, and the improvement rate was measured. Results:All the patients were followed up for (6.1±3.6)months (range, 3-9 months). The operation time was (55.1±8.2)minutes (range, 40-75 minutes), and intraoperative blood loss was (45.2±5.3)ml (range, 40-80 ml). The Zygapophysial joint reduction took (2.1±0.5)minutes (range, 1.5-3.0 minutes), with a success rate of 100%. Surgical procedures were performed with no postoperative complications such as aggravated spinal cord injury, large vascular injury or esophageal lesion. All the patients obtained Class I incision healing at first stage. The height of cervical intervertebral space was improved from preoperative (3.3±0.6)mm to (4.9±0.8)mm at postoperative 3 months ( P<0.05). The Cobb angle was increased from preoperative (-4.6±3.6)° to (6.5±2.1)° at postoperative 3 months ( P<0.01). According to Lenke classification, the intervertebral body fusion was good at postoperative 3 months, including Grade A in 41 patients and Grade B in 4. The rest of the patients showed varying degrees of neurological recovery according to ASIA scale, except for 6 patients with ASIA Grade A. The JOA score was improved from preoperative (7.4±2.3)points to (15.0±3.2)points at postoperative 3 months ( P<0.05), with the improvement rate of (73.3±17.6)%. Conclusion:For stage I anterior cervical reduction, decompression, interbody fusion and internal fixation of single-segment lower cervical injury of AO type C subtype F4, early surgical decompression is needed so as to reduce the perched facet or dislocated zygapophyseal joints and effectively improve the cervical spinal cord function.

Efficacy comparison of long/short segment posterior instrumentation combined with transpedicular impaction bone grafting in treating stage III Kümmell's disease / 中华创伤杂志

Objective:To compare the clinical efficacy of long/short segment posterior instrumentation combined with transpedicular impaction bone grafting for stage III Kümmell's disease.Methods:A retrospective case control study was conducted to analyze the clinical data of 45 patients with stage III Kümmell's disease admitted to Zhengzhou Orthopaedics Hospital from June 2012 to June 2019, There were 8 males and 37 females, aged 58-84 years [(68.8±3.5)years]. Segment of injury was T 11 in 5 patients, T 12 in 18, L 1 in 15, and L 2 in 7. A total of 24 patients were treated with posterior long segment pedicle screw fixation combined with pedicle screw compression and bone grafting (long segment group), and 21 patients were treated with posterior short segment pedicle screw fixation combined with pedicle screw compression and bone grafting (short segment group). For patients with severe osteoporosis, pedicle screws were augmented with bone cement. The operation time and blood loss were compared between the two groups. The visual analogue scale (VAS) was used to evaluate the degree of low back pain, and the Japanese Orthopaedic Association (JOA) score was used to evaluate the lumbar function at postoperative 2 weeks and 1 year. According to X-ray film and CT examination, the bone healing of the grafted vertebral body and the changes of Cobb angle of injured vertebrae were further evaluated. The complications were observed. Results:All patients were followed up for 15-48 months[(31.2±2.3)months]. There were no significant differences between the two groups in operation time and intraoperative blood loss ( P>0.05). The bone grafts in the vertebrae healed well in both groups at 1 year after operation. The Cobb angle, VAS and JOA score in both groups improved at 2 weeks and 1 year after operation ( P<0.01). There was no significant difference in Cobb angle, VAS and JOA score between the two groups at 2 weeks after operation ( P>0.05). The Cobb angle in short segment group [(14.8±6.3)°] was significantly higher than that in long segment group [(8.5±3.3)°] at 1 year after operation ( P<0.01), but there was no significant difference in VAS and JOA scores between the two groups ( P>0.05). There was no loosening or breakage of internal fixation in both groups. Conclusions:For stage III Kümmell's disease, both long and short segment posterior instrumentation combined with transpedicular impaction bone grafting can effectively restore the vertebral height, improve kyphosis and reduce dysfunction. However, long segment can better maintain spine stability, prevent vertebral collapse and progression of local kyphosis Cobb angle when compared with short segment fixation.

Anti-IFN-γ autoantibodies underlie disseminated Talaromyces marneffei infections.

Talaromyces marneffei causes life-threatening opportunistic infections, mainly in Southeast Asia and South China. T. marneffei mainly infects patients with human immunodeficiency virus (HIV) but also infects individuals without known immunosuppression. Here we investigated the involvement of anti-IFN-γ autoantibodies in severe T. marneffei infections in HIV-negative patients. We enrolled 58 HIV-negative adults with severe T. marneffei infections who were otherwise healthy. We found a high prevalence of neutralizing anti-IFN-γ autoantibodies (94.8%) in this cohort. The presence of anti-IFN-γ autoantibodies was strongly associated with HLA-DRB1*16:02 and -DQB1*05:02 alleles in these patients. We demonstrated that adult-onset acquired immunodeficiency due to autoantibodies against IFN-γ is the major cause of severe T. marneffei infections in HIV-negative patients in regions where this fungus is endemic. The high prevalence of anti-IFN-γ autoantibody-associated HLA class II DRB1*16:02 and DQB1*05:02 alleles may account for severe T. marneffei infections in Southeast Asia. Our findings clarify the pathogenesis of T. marneffei infection and pave the way for developing novel treatments.

Lysine succinylation and SIRT5 couple nutritional status to glutamine catabolism.

The metabolic microenvironment of tumors is characterized by fluctuating and limited nutrient availability. To survive these conditions, cancer cell-intrinsic mechanisms sense and signal nutritional status. We describe how glutaminase (GLS) is destabilized by lysine succinylation and stabilized by the NAD+-dependent desuccinylase sirtuin 5 (SIRT5), coupling nutrient levels to metabolic flux.

SIRT5 stabilizes mitochondrial glutaminase and supports breast cancer tumorigenesis.

The mitochondrial enzyme glutaminase (GLS) is frequently up-regulated during tumorigenesis and is being evaluated as a target for cancer therapy. GLS catalyzes the hydrolysis of glutamine to glutamate, which then supplies diverse metabolic pathways with carbon and/or nitrogen. Here, we report that SIRT5, a mitochondrial NAD+-dependent lysine deacylase, plays a key role in stabilizing GLS. In transformed cells, SIRT5 regulates glutamine metabolism by desuccinylating GLS and thereby protecting it from ubiquitin-mediated degradation. Moreover, we show that SIRT5 is up-regulated during cellular transformation and supports proliferation and tumorigenesis. Elevated SIRT5 expression in human breast tumors correlates with poor patient prognosis. These findings reveal a mechanism for increasing GLS expression in cancer cells and establish a role for SIRT5 in metabolic reprogramming and mammary tumorigenesis.

Liver-Type Glutaminase GLS2 Is a Druggable Metabolic Node in Luminal-Subtype Breast Cancer.

Efforts to target glutamine metabolism for cancer therapy have focused on the glutaminase isozyme GLS. The importance of the other isozyme, GLS2, in cancer has remained unclear, and it has been described as a tumor suppressor in some contexts. Here, we report that GLS2 is upregulated and essential in luminal-subtype breast tumors, which account for >70% of breast cancer incidence. We show that GLS2 expression is elevated by GATA3 in luminal-subtype cells but suppressed by promoter methylation in basal-subtype cells. Although luminal breast cancers resist GLS-selective inhibitors, we find that they can be targeted with a dual-GLS/GLS2 inhibitor. These results establish a critical role for GLS2 in mammary tumorigenesis and advance our understanding of how to target glutamine metabolism in cancer.

Characteristics and Prognosis of Talaromyces marneffei Infection in Non-HIV-Infected Children in Southern China.

Knowledge about the clinical and laboratory characteristics and prognosis of Talaromyces marneffei infection in children is limited. A retrospective study was conducted on pediatric patients with disseminated T. marneffei infection in a clinical setting. Extracted data included demographic information (age and sex), clinical features, laboratory findings, treatment, and prognosis. Eleven HIV-negative children were enrolled. The male/female ratio was 8:3. The median age of onset was 17.5 months (3.5-84 months). The mortality rate in these children was 36.36% (4/11). Seven children had underlying diseases. All of the children had multiple immunoglobulin abnormalities and immune cell decline. Ten children received voriconazole treatment, and most of the children (7/10) had a complete response to therapy at primary and long-term follow-up assessment; only three children died of talaromycosis. One patient recovered from talaromycosis but died of leukemia. The child who received itraconazole treatment also showed clinical improvement. No adverse events associated with antifungal therapies were recorded during and after the treatment. Talaromycosis is an indicator disease for undiagnosed severe immunodeficiencies in children. Awareness of mycoses in children by pediatricians may prompt diagnosis and timely treatment. Voriconazole is an effective, well-tolerated therapeutic option for disseminated T. marneffei infection in non-HIV-infected children.

In Vitro Susceptibility of Berberine Combined with Antifungal Agents Against the Yeast Form of Talaromyces marneffei.

Talaromyces (Penicillium) marneffei can cause fatal disseminated infection in immunocompromised hosts. However, therapeutic strategies for the mycosis are limited. Reports of the other fungi suggest that berberine, a component of traditional herb, inhibitors interact with antifungal agents to improve the treatment outcomes. In the study, we evaluated the in vitro efficacy of berberine in combination with conventional antifungal agents against the pathogenic yeast form of T. marneffei. We demonstrate the synergistic effect of combination of berberine with fluconazole (52.38%), itraconazole (66.67%), voriconazole (71.43%), amphotericin B (71.43%) or caspofungin (52.38%) of T. marneffei strains, respectively. Time-kill curves confirmed the synergistic interaction, and no antagonistic was observed in all of the combinations. In conclusion, berberine could enhance the efficacy of conventional antifungal agents against the yeast form of T. marneffei in vitro. The results indicated berberine might have a potential role in combination therapy for talaromycosis.

Subdivision of Brillouin gain spectrum to improve the spatial resolution of a BOTDA system.

A pulse subdivision analysis method was developed to improve the spatial resolution of a conventional long pump pulse Brillouin optical time domain analysis (BOTDA) system. An exclusive photodetector was used to obtain the accurate energy distribution along the long pulse, based on which the long pulse was subdivided into several sub-pulses with certain energy weights. With these energy weights, the Brillouin spectrum generated by the long pulse was subdivided into equal numbers of sub-spectra. Each sub-spectrum could provide detailed sensing information about a fiber sub-segment related to a sub-pulse. Thus, the actual spatial resolution of the BOTDA system was determined by the sub-pulse instead of the long pulse. As a result, spatial resolution was increased by several times, depending on the subdivision multiples. The method was theoretically simulated and experimentally demonstrated. For experimental demonstration, the recognization capability of the melting point of two different fiber sections and the discrete strain distribution on a sensing fiber were respectively tested. For melting point recognization, thanks to five-multiple subdivision, a 1 m spatial resolution over 31 km sensing fiber was realized using a 50 ns pump pulse. For the strain sensing test, ten-multiple subdivision was performed to distinguish two 0.5 m stretched fiber sections with a 0.2 m interval using a 20 ns pump pulse. The spatial resolution is 0.2 m, which is a ten times' improvement compared with that before subdivision analysis. Due to its simplicity and cost-effectiveness, the method is believed to have extensive application prospects in distributed fiber sensing fields.