Neoadjuvant sintilimab plus chemotherapy in EGFR-mutant NSCLC: Phase 2 trial interim results (NEOTIDE/CTONG2104).

The clinical efficacy of neoadjuvant immunotherapy plus chemotherapy remains elusive in localized epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC). Here, we report interim results of a Simon's two-stage design, phase 2 trial using neoadjuvant sintilimab with carboplatin and nab-paclitaxel in resectable EGFR-mutant NSCLC. All 18 patients undergo radical surgery, with one patient experiencing surgery delay. Fourteen patients exhibit confirmed radiological response, with 44% achieving major pathological response (MPR) and no pathological complete response (pCR). Similar genomic alterations are observed before and after treatment without influencing the efficacy of subsequent EGFR-tyrosine kinase inhibitors (TKIs) in vitro. Infiltration and T cell receptor (TCR) clonal expansion of CCR8+ regulatory T (Treg)hi/CXCL13+ exhausted T (Tex)lo cells define a subtype of EGFR-mutant NSCLC highly resistant to immunotherapy, with the phenotype potentially serving as a promising signature to predict immunotherapy efficacy. Informed circulating tumor DNA (ctDNA) detection in EGFR-mutant NSCLC could help identify patients nonresponsive to neoadjuvant immunochemotherapy. These findings provide supportive data for the utilization of neoadjuvant immunochemotherapy and insight into immune resistance in EGFR-mutant NSCLC.

Dynamic 18 F-FDG PET/CT can predict the major pathological response to neoadjuvant immunotherapy in non-small cell lung cancer.

Major pathological response (MPR) is a potential surrogate for overall survival. We determined whether the dynamic changes in 18 F-labeled fluoro-2-deoxyglucose positron emission tomography/computed tomography (18 F-FDG PET/CT) were associated with MPR in patients receiving neoadjuvant immunotherapy. Forty-four patients with stage II-III non-small cell lung cancer (NSCLC) who received neoadjuvant immunotherapy and radical surgery were enrolled. Moreover, 18 F-FDG PET/CT scans were performed at baseline and within 1 week before surgery to evaluate the disease. All histological sections were reviewed to assess MPR. The detailed clinical features of the patients were analyzed. The reliability of the clinical variables was assessed in differentiating between MPR and non-MPR using logistic regression. Receiver-operating characteristic (ROC) curve analysis identified the SUVmax changes threshold most associated with MPR. Most of the patients were pathologically diagnosed with squamous cell carcinoma and received anti-PD-1 antibodies plus chemotherapy. The immunotherapy regimens included nivolumab, pembrolizumab, and camrelizumab. MPR was observed in more than half of lesions. Tumors with MPR had a higher decrease in the longest dimension on dynamic PET/CT than those without MPR. Furthermore, the decline in SUVmax was significantly different between MPR and non-MPR diseases, and MPR lesions had a prominent mean reduction in SUVmax. SUVmax reduction was independently associated with MPR in the multivariate regression. On ROC analysis, the threshold of SUVmax decrease in 60% was associated with MPR. Dynamic changes in SUVmax were associated with MPR. The tumors with MPR showed a greater PET/CT response than those without MPR. A SUVmax decrease of more than 60% is more likely to result in an MPR after receiving neoadjuvant immunotherapy.

[The effectiveness of different respiration models to the amplitude of waveform information in arterial blood gas].

Objective: The objective is to find the characteristics of arterial blood sample waveform in different respiration models. Methods: Six post-operative patients with normal heart function and negative Allen test, were 4 male and 2 female, (59.00±16.64)year, (71.67±0.37)kg, left ventricular ejection fraction(LVEF) (61.33±2.16)%, had been placed the arterial catheterization and central venous catheterization for continuous collecting arterial in 3 different kinds of respiration models: normal breathing, no breathing and deep breathing. We selected two breaths cycles of waveform from each patient for data calculations of magnitudes and time interval. Compare the adjacent highest and lowest values of patients to verify whether there are periodic wave-like signal changes in arterial and venous blood gas in the three breathing states. In addition, statistical t-test analysis was performed on the change amplitude of the periodic wave-like signal of the patient's arterial and venous blood gas to compare whether there is a difference. Results: The heart beat numbers for drawing blood into pipe were 15-16, and all covered more than 2 breathing cycles. There were significant changes of arterial PaO2 (i.e. the highest high values compare to the next lowest values, P<0.05) in three different breathing models(normal, no breathing and high breathing), the magnitudes of which were (9.96±5.18)mmHg, (5.33±1.55)mmHg and (13.13±7.55)mmHg, with (8.09±2.43)%, (5.29±2.19)% and (10.40±2.68)% from their mean respectively. PO2 in venous blood gas did not show wavy changes under normal breathing, 20 s breath holding and high tidal volume ventilation. The amplitudes were (1.63 ± 0.41) mmHg, (1.13 ± 0.41) mmHg and (1.31 ± 0.67) mmHg, which were (3.91 ± 1.22)%, (2.92 ± 1.12)%, (3.33 ± 1.81)%, respectively, which were significantly lower than that of arterial blood gas under the same state, but there was no significant difference between groups. Conclusion: With continuous beat-by-beat arterial blood sampling and ABG analyzing method in three different breathing models, We obtain a clear evidence of the biggest periodic parameters ABG waveform in high breathing models, which followed by normal breathing models, no breathing was the smallest, and the wave variation amplitude of venous oxygen partial pressure was not obvious in the three respiratory states, which implies the oscillatory information of the arterial blood with comes from the gas exchanging in the lung.

[The experimental evidence of waveform information in arterial blood gas by beat-by-beat sampling method in human body-the differences of waveform information between arterial and venous blood samples].

Objective: The arterial blood with the oscillatory information comes from the right heart system after gas exchanging in the lung. However, the evidence of the waveform of venous ABG is lack. The objectives of this article are to compare the different information between arterial and venous beat-by-beat blood sample at the same time. Methods: Six post-operative patients with normal heart function and negative Allen test, had been placed the arterial catheterization and central venous catheterization directly connected to pre-heparin plasticpipes for continuous collecting arterial and venous blood. We twisted the 2 pipes into helix formation. After drawing arterial and venous blood with syringes in one heart beat with one helix at the same time, totally 15 heart beats, clipping the pipes with forceps, we put the helix pipe into icedwater at once and analyses PaO2, PaCO2, pH and SaO2 as soon as possible. We selected two breathscycles of waveform from each patient for data calculations of magnitudes and time interval. Results: The heart beat numbers for drawing blood into pipe were 15~16, and all covered more than 2 breathing cycles. There were significant changes of arterial PaO2(i.e. the highest high values compare to the next lowestvalues, P<0.05), but no significant changes in venous blood(P>0.05). The magnitudes of changing PaO2 in arterial and venous blood sample were (9.96±5.18)mmHg and (1.63±0.41)mmHg with significant variance(P=0.010), and they were (8.09±2.43)% and (3.91±1.22)%from their mean with significant variance(P=0.009) respectively. Conclusion: With continuous beat-by-beat arterial and venous blood sampling and ABG analyzing method at the same time, we obtain a clear evidence of periodic parameters ABG waveform, which following breathing cycle, but no clear ABG waveform of the periodic parameters in the venous blood samples, which implies the oscillatory information of the arterial blood with comes from the gas exchanging in the lung.

[Evidence of waveform information in arterial blood gas by beat-by-beat sampling method in patients with normal heart function].

OBJECTIVE: Since 2011 EB-APS conference, we hypotheses that phase switching of inspiration-expiration is dominantly initiated by oscillatory information PaO2, PaCO2 and [H+] via fast peripheral chemical receptors. However, the evidence of the waveform of ABG is lack. METHODS: Six surgery patients with normal heart function and negative Allen test, had been placed the arterial catheterization directly connected to 3 x 1 000 mm pre-heparin plastic pipe for continuous collecting arterial blood. We counted the number of heart beat for the blood collecting time, and separated the blood pipe into the heart beat numbers' short pieces using haemostatic forceps, then put pipe into iced water at once fir analyzing PaO2, PaCO2, pH and SaO2 as soon as possible. We selected two breaths cycles of waveform from each patient for data calculations of magnitudes and time interval. RESULTS: The heart beat numbers for filling blood into pipe were 16 ± 2, and all covered more than 2 breathing cycles. Each breathing cycle is cover 5 ± 0.6 heart beat. There were significant changes of PaO2, PaCO2, [H+] a and SaO2 (i.e. the highest high values compare to the next lowest values, P < 0.05). The time interval of changing PaO2, PaCO2, [H+]a and SaO2 magnitudes were 11.28 ± 1.13 mmHg, 1.77 ± 0.89 mmHg, 1.14 ± 0.35 nmol/L and 0.52% ± 0.44% respectively. CONCLUSION: This simple continuous beat-by-beat arterial blood sampling and ABG analyzing method is new and practicable. We obtain a clear evidence of periodic parameters ABG waveform, which following breathing cycle.

[Evidence of waveform information in arterial blood gas by beat-by-beat sampling method in patients with heart failure].

OBJECTIVE: We investigate the magnitudes of waveform changes of arterial blood gas (ABG) in patients with heart failure. METHODS: Five patients with heart failure were selected, continuous collecting radial artery blood and measured PaO2, PaCO2, pHa and Sao2. We selected two typical breaths cycles of waveform changes of ABG from each patient for data analysis. Comparison of the adjacent highest and lowest values to verify the presence of a periodic waveform changes of ABG, and in addition, we used t test to analysis the range of waveform changes of ABG in patients with heart failure and patients with normal cardiac function and compared whether the difference between them. RESULTS: The 5 patients (2 surgical and 3 ICU) with heart failure, were 4 male and 1 female, (69 ± 7)year, (169 ± 10) cm, (75 ± 19)kg, LVEF = (38 ± 3)%. The heart beat numbers for full blood into the blood sampling pipe were 17 ± 2, and all covered more than 2 breath cycles. There were significant changes of PaO2, PaCO2, [H+]a and SaO2 (P < 0.05). The magnitudes of changing PaO2, PaCO2, [H+]a and Sao2 were (7.94 ± 2.02)mmHg, (1.18 ± 0.56)mmHg, (0.54 ± 0.17)nmol/L and (0.21 ± 0.07)%, and they were (6.1 ± 1.5)%, (3.2 ± 1.5)%, (1.5 ± 0.5)% and (0.2 ± 0.1)% from their mean respectively. Even these magnitudes fo all ABG parameters were trendily lower than those of patients with normal cardiac function, but only PaO2 and [H+]a were significant (P < 0.05). CONCLUSION: Using this simple continuous beat-by-beat arterial blood sampling method, we obtained a clear evidence of periodic waveform of ABG parameters following by breath cycle in patients with heart failure, but the magnitude trendily be decreased.

[Effect of bone morphogenetic protein-7 on monocyte chemoattractant protein-1 induced epithelial-myofibroblast transition and TGF-beta1-Smad 3 signaling pathway of HKC cells].

OBJECTIVE: To examine the effect of Bone Morphogenetic protein-7 (BMP-7) on Monocyte chemoattractant protein-1 (MCP-1) induced epithelial-myofibroblast transition (EMT) in cultured renal proximal tubular cells (HK-2) and the relationship between TGF-beta1-smad 3 expressions and MCP-1 induced EMT. METHODS: The cultured HK-2 cells were divided into six groups: a, negative control, b, treated with TGF-beta1 (5 ng/ml) as positive control, c, treated with MCP-1 (0.1, 1, 10, 50 ng/ml), d, treated with BMP-7 (0.1, 1, 10, 50 ng/ml), e. co-treated with MCP-1 (1 ng/ml) and MCP-1 neutralized antibody (1 ng/ml), f. co-treated with MCP-1 (1 ng/ml) and BMP-7 (50 ng/ml). alpha-Smooth Muscle Actin (alpha-SMA) mRNA expression of HK-2 cells was assessed with RT-PCR. Secretion of type I collagen was assessed with RT-PCR and ELISA, respectively. TGF-beta1 and Smad 3 expressions were assessed with Western blot. RESULTS: alpha-SMA mRNA expression significantly increased in HK-2 cells treated with MCP-1 (0.1, 1 ng/ml) compared with negative controls (5.97 +/- 0.35, 23.36 +/- 1.37 vs. 0.59 +/- 0.38, P < 0.01). alpha-SMA mRNA expression of HK-2 cells concomitantly treated with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) significantly decreased than that in cells treated with MCP-1 (1 ng/ml) alone (1.93 +/- 0.34, 13.59 +/- 0.38 vs. 36.36 +/- 1.37, P < 0.01). Secretion of type I collagen of the cells treated with MCP-1 (0.1, 1 ng/ml) markedly increased compared with negative control (1751 +/- 34, 1876 +/- 45 vs. 1450 +/- 62; P < 0.01). The secretion of type I collagen of the supernatant were also significantly lower than that in cells treated with MCP-1 (1 ng/ml) alone (1462 +/- 56, 1596 +/- 34 vs. 1876 +/- 45, P < 0.05). The expression of TGF-beta1 and Smad 3 of HK-2 cells treated with MCP-1 (1 ng/ml) were markedly higher than that of negative controls, respectively (36.31 +/- 1.37 vs. 0.75 +/- 0.16, P < 0.01; 56.98 +/- 2.61 vs. 23.05 +/- 1.82, P < 0.01). The expressions of TGF-beta1 and Smad 3 in HK-2 cells treated concomitantly with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) were markedly decreased than that treated with MCP-1 alone, respectively. (4.61 +/- 0.74, 23.74 +/- 2.14 vs. 36.31 +/- 1.37, P < 0.01; 19.63 +/- 1.65, 37.06 +/- 1.82 vs. 56.98 +/- 2.61, P < 0.01). The expressions of TGF-beta1 and Smad 3 in HK-2 cells treated concomitantly with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) were markedly decreased than that treated with MCP-1 alone, respectively. (4.61 +/- 0.74, 23.74 +/- 2.14 vs. 36.31 +/- 1.37, P < 0.01; 19.63 +/- 1.65, 37.06 +/- 1.82 vs. 56.98 +/- 2.61, P < 0.01). CONCLUSIONS: The results documented that MCP-1 may induce EMT of HK-2 cells in vitro, and this effect is related to up-regulated expression of TGF-beta1 and Smad 3. BMP-7 may partially inhibit MCP-1-induced EMT and this effect is related to the downregulated expression of TGF-beta1 and Smad 3 of the cells. The results also suggest that MCP-1 induced EMT may involve the TGF-beta1-independent pathway of the cells.

[Altered expression of vascular endothelial growth factor and its receptors in transdifferentiated human proximal tubular epithelial cells induced by transforming growth factor beta1].

OBJECTIVE: To examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1). METHODS: The transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot. RESULTS: The immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners. CONCLUSIONS: Increased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

[Effect of BMP-7 on the transdifferentiation of cultured human tubular epithelial cell induced by TGF-beta1].

OBJECTIVE: To observe the effect of bone morphogenetic protein-7 (BMP-7) on the transdifferentiation of cultured human tubular epithelial cell (HKC) induced by TGF-beta1 and to elucidate its possible mechanism. METHODS: The cultured HKC cells were divided into 5 groups: serum-free group (negative control); single TGF-beta1 treated group (positive control); single BMP-7 treated group; combined TGF-beta1 and BMP-7 treated group; and BMP-7 pre-treated group. Expression of keratin of HKC cells was assessed by indirect enzyme immunohistochemistry (IEI), expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin by immunohistological method, percentage of alpha-SMA positive HKC cells by flow cytometry, and mRNA expression of alpha-SMA, TGF-beta1, and TGF-beta type II receptor by reverse transcription PCR. RESULTS: The expression of alpha-SMA and the percentage of alpha-SMA positive HKC cells markedly increased after having been treated by TGF-beta1 while the expression of E-cadherin and keratin decreased. In the group pre-treated with BMP-7 (50 ng/ml) and then added with TGF-beta1 (8 ng/ml), expression of alpha-SMA was significantly lower than in the positive control group, while expression of E-cadherin and keratin significantly higher than in the positive control group. Measurement of the percentage of alpha-SMA positive HKC found significant deference between the combined TGF-beta1 and BMP-7 treated group and the positive control group (9.7% vs 19.8%; 5.8% vs 19.8%; P < 0.05). Significant difference existed between the BMP-7 (50 ng/ml) pre-treated group and the positive control group (8.7% vs 19.8%, P < 0.05). mRNA expression of alpha-SMA was measured by RT-PCR and the results showed that it significantly decreased in the group treated or pre-treated with BMP-7 (50 ng/ml) (15% and 12% of the results in the positive control group, respectively). The mRNA expression levels of both TGF-beta1 and its type II receptor significantly decreased (28% and 19%; 47% and 36%, compared with the positive control group, respectively). CONCLUSION: Transdifferentiation of cultured renal epithelial cell induced by TGF-beta1 can be inhibittd by certain levels of BMP-7, cultured together with TGF-beta1 or pretreated. BMP-7 can prevent and inhibit the mRNA expression of TGF-beta1 and its type II receptor, which may be an important mechanism by which BMP-7 inhibit the transdifferentiation of renal tubular epithelial cell.