Changes of Mycobacterium tuberculosis specific antigen-stimulated CD27-CD38+IFN-γ+CD4+ T cells before and after anti-tuberculosis treatment.

BACKGROUND: The aim of the study was to investigate whether the expression of CD27-CD38+ in interferon (IFN)-γ+CD4+ T cells stimulated by the specific antigen early secreted antigenic target-6 (ESAT-6)/culture filter protein-10 (CFP-10) could be a potential new therapeutic evaluation indicator for anti-tuberculosis (TB) treatment. METHODS: Newly diagnosed active pulmonary TB patients, latent TB infection (LTBI) and healthy controls were enrolled from January 2021 to December 2021. PTB patients were treated by standard anti-TB regimen 2HREZ/4HR (2 months of isoniazid (H), rifampin (R), ethambutol (E), and pyrazinamide (Z) followed by 4 months of isoniazid (H) and rifampin (R)). The difference of CD27-CD38+ expression in IFN-γ+CD4+ T cells before treatment, 2 months after treatment, and 6 months after treatment were compared. RESULTS: Total 45 PTB patients, 38 LTBI cases and 43 healthy controls were enrolled. The expression of CD27-CD38+ decreased significantly after anti-TB treatment and was comparable with that in LTBI and healthy controls when the 6-month anti-TB treatment course was completed. The decline rate of CD27-CD38+ between 6 months after treatment and baseline was positively correlated with erythrocyte sedimentation rate (r = 0.766, P < 0.0001), C-reactive protein (r = 0.560, P = 0.003) and chest computerized tomography severity score (r = 0.632, P = 0.0005). The area under receiver operator characteristic curve of CD27-CD38+ in distinguish pulmonary TB patients before and after treatment was 0.779. CONCLUSION: The expression of CD27-CD38+ in ESAT-6/CFP-10 stimulated IFN-γ+CD4+T cells can well reflect the changes of the disease before and after anti-TB treatment, which is expected to be a potential new therapeutic evaluation index. Clinical Registry number chiCTR1800019966.

Eco-friendly approach for enhancing the floatability of non-metallic components in waste printed circuit boards: Adding gutter oil during dry grinding.

Waste printed circuit boards (WPCBs) are an attractive secondary resource that is challenging to dispose of due to its complexity. Reverse flotation is an effective method to remove non-metallic particles (NMPs) to obtain metals from WPCBs. Nevertheless, the removal of NMPs is usually inadequate in the present flotation practice. Thus, to provide a clean approach to improve the removal efficiency of NMPs, the method of adding gutter oil during dry grinding process was adopted to enhance the hydrophobic sites on the surface of NMPs to improve the floatability. The surface morphology of NMPs was analyzed by SEM, the results show that the rough morphology inhibited the adhesion of bubbles, while water occupied the cracks and pores, making it challenging for collector adsorption, which result in unstable particle-bubble adhesion. The results of FTIR indicate that both NMPs and gutter oil have -CH3, -CH2, -C = O, -C-O functional groups, which promotes the adsorption of gutter oil on the surface of NMPs. The contact angle (CA) results show that the adsorption of gutter oil on the particle surface is conducive to the formation of enhanced CA. Furthermore, the flotation enhancement effect was verified by flotation kinetic experiments. The accumulated floats yield of NMPs conditioned by gutter oil during grinding is increased from 67.05% (NMPs without conditioning) to 95.02%, and the resin recovery is increased by 31.10%. It is demonstrated that dry grinding with gutter oil can strengthen the floatability of NMPs, which provides a potential approach to increase the flotation efficiency of WPCBs.

[Effects of electroacupuncture on NLRP3/Caspase-1/GSDMD axis and neurological function in rats with cerebral ischemic reperfusion].

OBJECTIVE: To investigate the neuroprotective effect of electroacupuncture (EA) at "Quchi" (LI 11) and "Zusanli" (ST 36) in the rats with cerebral ischemic reperfusion and the potential mechanism of microglia pyroptosis. METHODS: Sixty SD rats were randomly divided into a sham-operation group, a model group and an EA group, with 20 rats in each group. The Zea Longa method was employed to establish the rat model of the middle cerebral artery occlusion and reperfusion (MACO/R) in the left brain. In the EA group, since the 2nd day of modeling, EA was given at "Quchi" (LI 11) and "Zusanli" (ST 36) of right side with disperse-dense wave, 4 Hz/20 Hz in frequency and 0.2 mA in current intensity, 30 min each time, once a day for lasting 7 consecutive days. The reduction rate of cerebral blood flow was measured with laser Doppler flowmetry during operation. The neurological function of rats was observed using Zea Longa neurobehavioral score. The cerebral infarction volume was detected by TTC staining method. The microglia positive expression in the ischemic side of the cortex was detected with the immunofluorescence method. Under transmission electron microscope, the ultrastructure of cell in the ischemic cortex was observed. The mRNA expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), cysteinyl aspartate specific proteinase-1 (Caspase-1) and gasdermin D (GSDMD) in the ischemic cortex were detected using real-time PCR. RESULTS: Compared with the sham-operation group, in the model group, the reduction rate of cerebral blood flow was increased during operation (P<0.001); Zea Longa neurobehavional score and the percentage of cerebral infarction volume were increased (P<0.001), the numbers of M1-type microglia marked by CD68+ and M2-type microglia marked by TMEM119+ were elevated in the ischemic cortex (P<0.001), the mRNA expression of NLRP3, ASC, Caspase-1 and GSDMD was increased (P<0.001, P<0.01); the cytomembrane structure was destroyed, with more cell membrane pores formed in the ischemic cortex. Compared with the model group, after intervention, Zea Longa neurobehavioral score and the percentage of cerebral infarction volume were reduced (P<0.05), the number of M1-type microglia marked by CD68+ was reduced (P<0.05) and the number of M2-type microglia marked by TMEM119+ was increased (P<0.05); and the mRNA expression of NLRP3, ASC, Caspase-1 and GSDMD was decreased (P<0.01, P<0.05) in the EA group. Even though the cytomembrane structure was incomplete, there were less membrane pores presented in the ischemic cortex in the EA group after intervention. CONCLUSION: The intervention with EA attenuates the neurological dysfunction and reduces the volume of cerebral infarction in the rats with cerebral ischemic reperfusion. The underlying mechanism is related to the inhibition of microglia pyroptosis through modulating NLRP3/Caspase-1/GSDMD axis.

High value of early secretory antigenic target-6 and culture filtrate protein-10 in diagnosis of IgA nephropathy with renal tuberculosis: a retrospective study.

BACKGROUND: This retrospective study aimed to evaluate the diagnostic value of early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) in immunoglobulin A nephropathy (IgAN) associated with renal tuberculosis (RT). METHODS: Forty patients with IgAN (IgAN group), 32 patients with RT (RT group), and 52 patients with IgAN associated with RT (IgAN + RT group) were retrospectively selected for this study. A tuberculin skin test (TST) was conducted, and Mycobacterium tuberculosis (MTB) antibody levels were measured. Immunohistochemistry and western blotting were used to determine the expression of ESAT-6 and CFP-10 proteins in renal tissues. RESULTS: The positive results of TST and levels of serum and urinary MTB antibodies were higher in the RT group than in the IgAN + RT group. The expression levels of ESAT-6 and CFP-10 proteins were the highest in the IgAN + RT group and lowest in the IgAN group. The receiver operating characteristic curves indicated that the area under curve (AUC) value of the ESAT-6 protein for the diagnosis of IgAN associated with RT was 0.907 and the cut-off value of the integral optical density (IOD) was 26.72. Diagnosis based on ESAT-6 protein levels showed 75% sensitivity and 94.2% specificity. The AUC value of the CFP-10 protein for the diagnosis of IgAN associated with RT was 0.8 and the cut-off value of IOD was 25.67. Detection based on CFP-10 protein levels showed 63.9% sensitivity and 84.6% specificity. CONCLUSIONS: Our study provides evidence for the potential of ESAT-6 and CFP-10 proteins as candidate markers in the diagnosis of IgAN associated with RT.

[Electroacupuncture promotes the repair of neurological function in ischemic stroke rats by inhi-biting cofilin rod formation].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on modified neurological severity score (mNSS), cerebral infarction volume, expression of Lim domain kinase-1 (LIMK1) and slingshot homolog-1 (SSH1) proteins, Cofilin rod formation and neural cell apoptosis in rats with ischemic stroke (IS), so as to explore its mechanisms underlying improvement of IS. METHODS: Male SD rats were randomly divided into normal, model and EA groups, with 13 rats in each group. The IS model was established by occlusion of the middle cerebral artery (MCAO) according to Zea Longa's method. EA was applied to "Quchi" (LI11) and "Zusanli" (ST36) for 30 min, once a day for 7 consecutive days. The behavioral changes of mNSS were observed before and after modeling. The volume of cerebral infarction was measured by using a small animal magnetic resonance imaging. The protein expressions of LIMK1 and SSH1 in the cerebral ischemic tissues were detected by Western blot. The density of Cofilin rod and neural cell apoptosis in cerebral ischemic area were determined by immunofluorescence staining and TUNEL staining, separately. RESULTS: After modeling, the mNSS score, cerebral infarction volume ratio, expression level of SSH1, density of Cofilin rod and the number of apoptotic cells were significantly increased (P<0.01), while the expression level of LIMK1 protein was obviously decreased in the model group relevant to the normal group (P<0.01). After 7 days' treatment, all the increased and decreased levels of the indexes mentioned above were reversed in the EA group relevant to the model group (P<0.01, P<0.05). CONCLUSION: EA of LI11 and ST36 can improve neurological function and reduce infarction range in MCAO rats, which may be related to its action in regulating the expression of LIMK1 and SSH1, inhibiting the formation of Cofilin rod and reducing apoptosis of neural cells.

Electroacupuncture Attenuates Ischemic Brain Injury and Cellular Apoptosis via Mitochondrial Translocation of Cofilin.

OBJECTIVE: To investigate the potential mechanisms of electroacupuncture (EA) to prevent ischemic stroke. METHODS: The method of middle cerebral artery occlusion (MCAO) was employed to establish a rat model of ischemic stroke. Seventy-eight Sprague-Dawley rats were divided into the sham group, MCAO + EA control (EC) group, and MCAO + EA (EA) group according to a random number table (n=26 per group). EA was applied to the acupoints of Baihui (DU 20) and Shenting (DU 24) 5 min and 6 h, respectively after the onset of MCAO. Rats in the sham and EC groups received only light isoflurane anesthesia for 30 min after MCAO. The neuroprotective effects of EA were evaluated by rota-rod test, neurological deficit scores and infarct volumes. Additionally, Nissl staining and immunostaining were performed to examine brain damage, rod formation, cellular apoptosis, and neuronal loss induced by ischemia. The activities of caspase-3, and expression levels of cofilin and p-cofilin in mitochondria and cytoplasm after ischemic injury were determined by Western blot. RESULTS: Compared with the EC group, EA significantly improved neuromotor function and cognitive ability after ischemic stroke (P<0.05 or P<0.01). Therapeutic use of EA also resulted in a significant decrease of cofilin rod formation and microtubule-associated protein-2 (MAP2) degradation in the cortical penumbra area compared with the EC rats (P<0.01). Furthermore, Western blot analysis showed that EA stimulation significantly inhibited mitochondrial translocation of cofilin and caspase-3 cleavage (P<0.05 or P<0.01). Additionally, brain damage (infarct volume and neuropathy), cellular apoptosis and neuronal loss induced by ischemia were remarkably suppressed by EA in the cortical penumbra of rats (P<0.05 or P<0.01). CONCLUSION: EA treatment after ischemic stroke may attenuate ischemic brain injury and cellular apoptosis through the regulation of mitochondrial translocation of cofilin, a novel mechanism of EA therapy.

Anti-proliferation and radiosensitization effects of chitooligosaccharides on human lung cancer line HepG2.

OBJECTIVE: To observe the anti-proliferation and radiosensitization effect of chitooligosaccharides (COS) on human lung cancer cell line HepG2. METHODS: CCK-8 assay was employed to obtain the inhibition ratio of COS on HepG2 cells at 24 h after treatment. The clonogenic assay was used to analyze the cell viability of RAY group and RAY + COS group with X-ray of 0, 1, 2, 4, 6 and 8 Gy, and the cell survival curve was used to analyze the sensitization ratio of COS. Flow cytometry was employed to detect cell cycle and apoptosis rate in control group, RAY group and RAY + COS group after 24 h treatment. RESULTS: COS inhibited the proliferation of HepG2 cells, and the inhibition rate positively correlated with the concentration of COS. The cell viability decreased with increasing exposure dose in RAY group and RAY + COS group. The cell viabilities of RAY + COS group were lower than those of RAY group at the dose of 4, 6 and 8 Gy (P < 0.05), and the sensitization ratio of COS was 1.19. There were higher percentage at G2/M phase and apoptosis rate, and lower percentage at S phase in RAY + COS group versus the other two groups (P < 0.01). CONCLUSIONS: COS can inhibit the proliferation of HepG2 cells, and enhance the radiosensitization of HepG2 cells, induce apoptosis and G2/M phase arrest.

Social defeat stress promotes tumor growth and angiogenesis by upregulating vascular endothelial growth factor/extracellular signal-regulated kinase/matrix metalloproteinase signaling in a mouse model of lung carcinoma.

Numerous epidemiological and experimental animal studies have indicated that chronic psychological stress may promote tumor development. However, the underlying molecular mechanisms by which chronic stress promotes tumorigenesis remain to be fully elucidated and animal models have not yet been well established. In the present study, an established mouse model of repeated social defeat stress (RSDS), was generated and used to investigate the effect of stress on tumor growth and metastasis. C57BL/6 mice were exposed to RSDS for 10 days, followed by subcutaneousl inoculation with Lewis lung carcinoma cells for seven days. The tumor weight and volume as well as the number of the lung metastatic nodules were then determined. Vascular endothelial growth factor (VEGF) serum levels were measured using ELISAs. In addition, expression levels of VEGF receptor (VEGFR) and L1 cell adhesion molecule (L1CAM) messenger (m)RNA were confirmed using reverse transcription quantitative polymerase chain reaction. Furthermore, protein expression levels of phosphorlyated extracellular signal-regulated kinase (pERK), matrix metalloproteinase (MMP)-2 and MMP-9 were examined using western blot analysis. The results showed that RSDS significantly increased the weight and the volume of the primary tumor as well as the number of the lung metastatic nodules. Serum VEGF levels were significantly higher in the tumor-stress group compared with those of the unstressed tumor mice. In addition, tumors in stressed animals demonstrated markedly enhanced expression of VEGFR-2 and L1CAM mRNA as well as pERK, MMP-2 and MMP-9 protein expression. In conclusion, these results suggested that RSDS contributed to lung cancer progression, angiogenesis and metastasis, which was partially associated with increased VEGF secretion and therefore the activation of the ERK signaling pathway, resulting in the induction of MMP-2 and MMP-9 protein expression.

[Clinical analysis of 81 cases of pulmonary cryptococcosis].

OBJECTIVE: To clarify the clinical feature, diagnosis and therapy of the pulmonary cryptococcosis (PC). METHODS: A retrospective study of cases with PC who were diagnosed by pathological examinations between January 1996 and December 2010 was conducted. Eighty-one cases were enrolled in the study (58 male and 23 female patients; mean age of (51±11) years). Forty-one cases were asymptomatic at the time of diagnosis. There were single pulmonary lesions in 50 cases, and multiple lesions in 31 cases. Fourteen lesions (17.3%) were located in left upper lobe, 27 (33.3%) in left lower lobe, 21 (25.9%) in right upper lobe, 3 (3.7%) in right middle lobe, 28 (34.6%) in right lower lobe, and 3 (3.7%) diffusely involved bilateral lungs. The tumors ranged from 0.8 to 10.0 cm in diameter with a mean of (2.9±1.8) cm. All the cases were misdiagnosis prior to the surgical resection, and histologically confirmed by postoperative pathological specimens. RESULTS: All the cases received surgical treatment including complete resection in 69 cases, and palliative resection in 12 cases. Resections were performed by means of video-assisted thoracoscopy in 31 cases and thoracotomy in 50 cases. Surgical resections included pulmonary wedge excisions in 42 cases, and lobectomies in 39 cases. After histological confirmation, 63 cases (77.8%) were treated with antifungal agents, which consisted of fluconazole in 38 cases, itraconazole in 18 cases, amphotericin B in 6 cases, and flucytosine in 4 cases. There were no intraoperative death, but two cases died for cryptococcal meningoencephalitis in the postoperative period. Operative morbidity occurred in 7 (8.6%) cases. The median follow-up was 42.5 months (6 to 84 months). There were 2 local relapses of PC, and 9 cases with complications of anti-fungal agents. CONCLUSIONS: The clinical manifestations of PC are mild and non-specific, with no characteristic radiographic manifestations. Surgical resection is usually indicated for definite diagnosis and treatment. Antifungal drug therapy is indispensable even after complete resection.

[Surgery treatment for pulmonary sclerosing hemangioma].

OBJECTIVE: To investigate the clinicopathological features and surgical treatment of pulmonary sclerosing hemangioma (PSH). METHODS: Clinic data of PSH patients admitted by surgical resection from January 1985 to December 2010 was analyzed retrospectively. One hundred and sixty-five patients were enrolled in the study. There were 27 male and 138 female patients with a mean age of (48 ± 13) years. Seventy-nine patients were asymptomatic at the time of diagnosis. Eighty-nine tumors arose in the right lung (27 in right upper lobe, 24 in right middle lobe, 34 in right lower lobe, 2 in right upper lobe with invasion of right middle lobe, 1 in right middle lobe with invasion of right lower lobe, and 1 case with multiple lobe lesions), 75 in the left (33 in left upper lobe, 42 in left lower lobe), and 1 in the bilateral. There were huge mass lesions in 2 cases, endobronchial lesions in 2 cases, and multiple lesions in 6 cases. The mean size of the lesion was (2.6 ± 0.9) cm (ranging from 0.9 to 10.0 cm). Forty-eight cases (29.1%) were misdiagnosed as malignancies preoperatively, and 41 cases (24.8%) were misdiagnosed intraoperatively. RESULTS: Resections were performed by means of video-assisted thoracoscopy (n = 53) and thoracotomy (n = 112). Surgical resection included pulmonary wedge excision in 61 patients, lobectomy in 89 patients, right bilobectomy in 5 patients, anatomic segmentectomy in 2 patient, enucleation in 6 patients, and synchronous bilateral pulmonary wedge resection in 1 patient. Operative mortality and morbidity occurred in 0 and 2 (4.3%) patients, respectively. Mean follow-up was 34.7 months (ranging from 6 to 62 months). There was no local recurrence or death from PSH. CONCLUSIONS: PSH is a rare benign lung tumor. It is difficult to make accurate diagnosis preoperatively, and sometimes even intraoperative frozen sections can't differentiate it from malignant tumors. Surgical resection is usually indicated for definite diagnosis and treatment. Partial resection is a sufficient treatment in view of uncommon tumor recurrence. Thoracoscopic surgery is recommended for PSH.

Performance of PAHs emission from bituminous coal combustion.

Carcinogenic and mutagenic polycyclic aromatic hydrocarbons (PAHs) generated in coal combustion have caused great environmental health concern. Seventeen PAHs (16 high priority PAHs recommended by USEPA plus Benzo[e]pyrene) present in five raw bituminous coals and released during bituminous coal combustion were studied. The effects of combustion temperature, gas atmosphere, and chlorine content of raw coal on PAHs formation were investigated. Two additives (copper and cupric oxide) were added when the coal was burned. The results indicated that significant quantities of PAHs were produced from incomplete combustion of coal pyrolysis products at high temperature, and that temperature is an important causative factor of PAHs formation. PAHs concentrations decrease with the increase of chlorine content in oxygen or in nitrogen atmosphere. Copper and cupric oxide additives can promote PAHs formation (especially the multi-ring PAHs) during coal combustion.