[Value of surgery combined chemotherapy and radiation therapy in locally advanced neuroendocrine carcinoma of the cervix: a single-center retrospective cohort study].

Objective: To evaluate the surgery combined chemotherapy and radiation in locally advanced neuroendocrine carcinoma of the cervix (NECC) . Methods: This is a single-center retrospective cohort study. Locally advanced NECC patients admitted to Peking Union Medical College Hospital, Chinese Acadmy of Medical Sciences from January 2011 to April 2022 were enrolled. They were divided into concurrent chemoradiotherapy group, and surgery combined with chemotherapy and radiation group. The Kaplan-Meier method was used to analyze the progression free survival (PFS), overall survival (OS), recurrence rate, and mortality rate. Results: (1) Forty-six cases were included, 22 in concurrent chemoradiotherapy group, 24 in surgery combined chemotherapy and radiation group. With 16 patients (35%, 16/46) received neoadjuvant chemotherapy (NACT), the NACT effective rate was 15/16. (2) The median follow-up time was 27.5 months (range: 10-106 months), with 26 (57%, 26/46) experienced recurrences. There were 4 (9%, 4/46) pelvic recurrences and 25 (54%, 25/46) distant recurrences, and 3 (7%, 3/46) both pelvic and distant recurrences. Compared with concurrent chemoradiotherapy group, surgery combined chemotherapy and radiation group had lower pelvic recurrence rate [14% (3/22) vs 4% (1/24); χ2=1.296, P=0.255] but without statistic difference. Both groups had similar distant recurrence rate [55% (12/22) vs 54% (13/24); χ2=0.001, P=0.979] and overall recurrence rate [59% (13/22) vs 54% (13/24); χ2=0.113, P=0.736]. (3) During the follow-up period, 22 cases (48%, 22/46) died, with 11 cases (50%, 11/22) in concurrent chemoradiotherapy group and 11 cases (46%, 11/24) in surgery combined chemotherapy and radiation group, without significant difference (χ2=0.080, P=0.777). The postoperative 3-year and 5-year OS rates were 62.3% and 36.9%. Compared with concurrent chemoradiotherapy group, the patients in surgery combined chemotherapy and radiation group showed an extended trend in PFS (17.0 vs 32.0 months) and OS (37.0 vs 50.0 months) but without statistic differences (P=0.287, P=0.125). Both groups had similar 3-year OS rate (54.2% vs 69.9%; P=0.138) and 5-year OS rate (36.1% vs 38.8%; P=0.217). Conclusions: Our study supports the multi-modality treatment strategy (including surgery, chemotherapy and radiation) as an important component in the treatment of locally advanced NECC. The combination of surgery, chemotherapy and radiation seems to have advantages in the treatment of locally advanced NECC, but needs to be confirmed by further multicenter studies.

[Value of postoperative radiotherapy and analysis of prognostic factors in early-stage neuroendocrine carcinoma of cervix].

Objective: To evaluate the effect of postoperative radiotherapy and high-risk pathological factors on the prognosis of early-stage neuroendocrine carcinoma of cervix (NECC). Methods: A single-center retrospective cohort study of early-stage NECC in Peking Union Medical College Hospital from January 2011 to April 2022 were enrolled. The patients were treated with radical hysterectomy±adjuvant treatment. They were divided into postoperative non-radiation group and postoperative radiation group. The possible postoperative recurrence risk factors identified by univariate analysis were assessed using multivariate logistic regression. The Kaplan-Meier method was used to analyze the progression free survival (PFS), overall survival (OS), recurrence rate, and mortality rate. Results: (1) Sixty-two cases were included in the study, including 33 cases in postoperative non-radiation group and 29 cases in postoperative radiation group. (2) The median follow-up time was 37 months (ranged 12-116 months), with 23 cases (37%) experienced recurrences. There were 7 cases (11%) pelvic recurrences and 20 cases (32%) distant recurrences, in which including 4 cases (6%) both pelvic and distant recurrences. Compared with postoperative non-radiation group, the postoperative radiation group had a lower pelvic recurrence rate (18% vs 3%; P=0.074) but without statistic difference, a slightly elevated distant recurrence rate (24% vs 41%; P=0.150) and overall recurrence rate (33% vs 41%; P=0.513) without statistically significances. Univariate analysis showed that lymph-vascular space invasion and the depth of cervical stromal invasion≥1/2 were risk factors for postoperative recurrence (all P<0.05). Multivariate analysis showed lymph-vascular space invasion was an independent predictor for postoperative recurrence (OR=23.03, 95%CI: 3.55-149.39, P=0.001). (3) During the follow-up period, 18 cases (29%, 18/62) died with tumor, with 10 cases (30%, 10/33) in postoperative non-radiation group and 8 cases (28%, 8/29) in postoperative radiation group, without significant difference (P=0.814). The postoperative 3-year and 5-year survival rate was 79.2%, 60.8%. The depth of cervical stromal invasion≥1/2 was more common in postoperative radiation group (27% vs 64%; P=0.011), and postoperative radiation in such patients showed an extended trend in PFS (32.3 vs 53.9 months) and OS (39.4 vs 73.4 months) but without statistic differences (P=0.704, P=0.371). Compared with postoperative non-radiation group, the postoperative radiation did not improve PFS (54.5 vs 37.3 months; P=0.860) and OS (56.2 vs 62.4 months; P=0.550) in patients with lymph-vascular space invasion. Conclusions: Postoperative radiation in early-stage NECC patients has a trend to reduce pelvic recurrence but not appear to decrease distant recurrence and overall recurrence, and has not improved mortality. For patients with the depth of cervical stromal invasion≥1/2, postoperative radiation has a trend of prolonging OS and PFS but without statistic difference. Lymph-vascular space invasion is an independent predictor for postoperative recurrence, but postoperative radiation in such patients does not seem to have any survival benefits.

[Improvement effect of Lycium barbarum polysaccharide on the intestinal flora of pregnant rats and their offspring under chronic stress].

Objective: To understand the improvement effect of Lycium barbarum polysaccharide (LBP) on the intestinal flora of mother mice during pregnancy and their offspring who experienced chronic stress, and provide new ideas for improving the effect of stress on the intestinal tract. Methods: From July to October 2019, 24 SPF-grade female SD rats were selected and divided into control group, stress group, and stress+LBP group, with 8 rats in each group. A chronic unpredictable mild stimulation model during pregnancy was established (21 days) , and 40 mg/kg LBP solution was administered by gavage on the 8th day of stress. Venous blood from the medial canthus of the female mice was collected on the 1st day before stress and on the 1st, 7th, 14th and 21st days, respectively. Cortisol was measured and corticosterone concentration was calculated. The fresh feces of famale mice after stress and 20-day postnatal offspring mice were collected, and Illumina Miseq sequencing technology, alpha diversity and community composition were used to analyze the diversity and structure of intestinal flora. Results: On the 7th and 14th days of stress, the plasma corticosterone concentration of female mice in the stress group and stress+LBP group was higher than that in the control group (P<0.05) . In the Alpha diversity of female mice, the Ace index of the stress group was lower than that of the control group (P<0.05) . The analysis of intestinal flora structure showed that at the species level, the proportions of Lachnospiraceae and Lactobacillus in the stress+LBP group were higher than those in the stress group and control group. At the order level, the proportion of Clostridiales in the stress+LBP group was higher than that in the stress group and lower than that in the control group, while the proportion of Lactobacillales was higher than that in the stress group and control group. In the Alpha diversity of the offspring group, the Shannon index, Ace index and Chao index of the stress+LBP offspring group were higher than those of the stress offspring group (P<0.05) . The proportion of Lactobacillus in the stress+LBP offspring group was higher than that in the control offspring group and stress offspring group, and the proportions of Lachnospiraceae and Ruminococcaceae in the stress+LBP offspring group were higher than those in the stress offspring group, the proportion of Bacteroidales in the stress+LBP offspring group was lower than that in the stress offspring group, and the proportion of Clostridiales in the stress+LBP offspring group was higher than that in the stress and control offspring groups. Conclusion: The intervention of LBP may improve the changes in the intestinal flora diversity, abundance and flora structure of mother mice and offspring caused by pregnancy stress, thereby maintaining the balance of intestinal flora.

LncRNA XIST promotes proliferation and cisplatin resistance of oral squamous cell carcinoma by downregulating miR-27b-3p.

Chemotherapy resistance has become a major obstacle to effective treatment of human cancer. This study aimed to investigate the effect of lncRNA XIST on cell proliferation and cisplatin (CDDP) of oral squamous cell carcinoma (OSCC). RT-qPCR and Western blot analysis were used to detect mRNA and protein expression. CCK-8 and flow cytometry assays were explored to evaluate CDDP sensitivity in OSCC cells. The relationship between lncRNA XIST and miR-27b-3p was confirmed by luciferase reporter assay. The results showed that lncRNA XIST was upregulated in OSCC tissues, cell lines, and CDDP-resistant OSCC cells. Functionally, upregulation of lncRNA XIST promoted cell proliferation, enhanced CDDP resistance, and inhibited apoptosis in OSCC cells. In addition, lncRNA XIST acts as a molecular sponge for miR-27b-3p in OSCC. Downregulation of miR-27b-3p partially reversed the tumor suppression effect and CDDP chemosensitivity of XIST knockdown in CDDP-resistant OSCC cells. In conclusion, lncRNA XIST promotes cell proliferation and enhances resistance to CDDP in OSCC by downregulating miR-27b-3p.

[A prospective cohort study about the screening tests of mismatch repair protein and clinical criteria for Lynch syndrome associated endometrial carcinoma].

Objectives: Currently, the commonly used screening methods for Lynch syndrome in patients with endometrial cancer (EC) are clinical diagnostic criteria and immunohistochemical testing. Our study compared the accuracy of the two methods in this prospective cohort study. Methods: Mismatch repair (MMR) protein was detected by immunohistochemical in the pathological tissues of newly diagnosed EC patients in Peking Union Medical College Hospital, during December 2015 and June 2018. Lynch syndrome related mutation gene was detected in patients with MMR protein deficiency. At the same time, all the patients were evaluated by the clinical diagnostic criteria (Amsterdam Criteria Ⅱ and the revised Bethesda criteria). Results: A total of 121 newly diagnosed EC patients were enrolled in this study, and 41 cases (33.9%) were MMR protein deficient. All of them received Lynch syndrome related mutation gene detection, and 7 cases were finally diagnosed with Lynch syndrome. Only 6 cases of Lynch syndrome, however, were diagnosed by the clinical diagnostic criteria, with 1 case misdiagnosed and 2 cases missed diagnosed. Conclusion: The incidence of Lynch syndrome in endometrial cancer patients is 5.8%. And the clinical diagnostic criteria for Lynch syndrome in patients with EC will result in miss diagnosis and misdiagnosis.

TLR2 regulates allergic airway inflammation through NF-κB and MAPK signaling pathways in asthmatic mice.

OBJECTIVE: To investigate the role of toll-like receptor 2 (TLR2) in asthmatic mouse model and its possible signal transduction pathways. MATERIALS AND METHODS: Mice were divided into three groups: TLR2-/- asthma mouse model group (n=10), C57BL/6 asthma mouse model group (n=10) and control group (n=10). Mice were sensitized and stimulated with ovalbumin (OVA) to establish the asthmatic mouse model. The unilateral bronchoalveolar lavage fluid (BALF) was collected and centrifuged to separate cells, and the cells were classified and counted via smear test under a microscope. Part of the lung tissues on the other side was taken for hematoxylin-eosin (HE) staining to observe the histopathological change in lung tissues. The remaining lung tissues on the other side were taken to detect the messenger ribonucleic acid (mRNA) expression levels of interleukin-4 (IL-4), IL-5 and IL-13 via reverse transcription-polymerase chain reaction (RT-PCR). The levels of nuclear factor-κB (NF-κB) p65, phosphorylated (p)-NF-κB p65, p-IκBα, extracellular signal-regulated kinase (ERK)1/2, p-ERK1/2, Jun N-terminal kinase (JNK), p-JNK, p38 mitogen-activated protein kinase (MAPK), p-p38 MAPK, IL-4, IL-5 and IL-13 were detected via enzyme-linked immunosorbent assay (ELISA). The protein expressions of NF-κB p65, p-NF-κB p65, p-IκBα, ERK1/2, p-ERK1/2, JNK, p-JNK, p38 MAPK, and p-p38 MAPK were detected using the immunohistochemical method. RESULTS: HE staining showed that the infiltration degree of inflammatory cells in perivascular tissues in TLR2-/- asthma group was reduced compared with that in C57BL/6 asthma group. Results of electron microscopy showed that the ultrastructural changes in alveolar type I epithelial cells in mice in TLR2-/- asthma group was significantly alleviated. In BALF in TLR2-/- asthma group, the numbers of eosinophils and lymphocytes were significantly decreased, but the number of macrophages was significantly increased compared with those in C57BL/6 asthma group. Results of RT-PCR and ELISA revealed that the mRNA and protein expression levels of IL-4, IL-5, and IL-13 in lung tissues of mice in TLR2-/- asthma group were significantly decreased compared with those in C57BL/6 asthma group. Besides, results of ELISA and immunohistochemistry revealed that the protein expressions of NF-κB p65, p-NF-κB p65, p-IκBα, ERK1/2, JNK, p38 MAPK, p-ERK1/2, p-JNK, and p-p38 MAPK in lung tissues of mice in TLR2-/- asthma group were significantly decreased compared with those in C57BL/6 asthma group. CONCLUSIONS: TLR2 is involved in the occurrence and development of experimental asthmatic airway inflammation. TLR2 gene knockout in asthmatic mice can alleviate the airway inflammation, whose mechanism may be that the allergic airway inflammation of asthmatic mice is alleviated through inhibiting NF-κB and MAPK signaling pathways.

The lack of Raf-1 kinase feedback regulation enhances antiapoptosis in cancer cells.

Raf-1 has an important role in cellular antiapoptosis. So far, there is no solid evidence that shows that Raf-1 mutation is associated with cancer development. In the course of further study of Raf-1 signaling, we have reported that Raf-1 hyperphosphorylation inhibits its kinase activity toward its downstream mitogen-activated protein kinase kinase 1/2 (MEK1/2) and proposed a model for negative feedback regulation of Raf-1. Here, we show that there is no hyperphosphorylation in some cancer cells, which results in increased kinase activity and enhances the antiapoptotic ability. Inhibition of either Raf-1 or ALG-2 (apoptosis-linked gene 2) expression results in apoptosis signal-regulating kinase 1/c-Jun N-terminal kinase (ASK1/JNK) signaling activation, and cell sensitivity to chemotherapeutic reagents, indicating that inhibition of ASK1/JNK apoptotic signaling by Raf-1 is mediated by ALG-2. A previous report indicated that extracellular signal-regulated kinase 1/2 (ERK1/2) were responsible for Raf-1 hyperphosphorylation. However, our evidence shows that when ERK1/2 are activated and the Raf-1 gene is not mutated, Raf-1 is not hyperphosphorylated in these cells, indicating that ERK1/2 are not responsible for the Raf-1 hyperphosphorylation in these cancer cell lines. Surprisingly, we also found that Raf-1 is not a necessary kinase for MEK1/2 activation under normal tissue culture conditions, but is required for MEK1/2 activation under apoptosis-inducing conditions. Our research demonstrates that although Raf-1 gene is not mutated, an abnormality of Raf-1 kinase feedback regulation enhances its antiapoptotic function, and Raf-1 can still be a pharmaceutical target to increase chemotherapy or radiotherapy sensitivity in these cancer cells.

Expression of platelet-derived growth factor B-chain and platelet-derived growth factor beta-receptor in fibroblasts of scleroderma.

Scleroderma is a fibrotic disease occurring in a localized or systemic form. The pathogenesis of scleroderma remains poorly understood. Recent studies revealed that various cytokines and growth factors were involved in the development of scleroderma fibrosis. Platelet-derived growth factor (PDGF) is a potent growth factor for mesenchymal cells, especially fibroblasts. It can promote fibroblasts proliferation, enhance extracellular matrix synthesis. It is also a chemoattractant to fibroblasts. To better understand the role of PDGF in pathogenesis of scleroderma, we performed both in vivo studies on the expression of PDGF beta-receptor protein in scleroderma tissue and in vitro studies on the expression of PDGF B-chain and PDGF beta-receptor mRNA in cultured fibroblasts derived from both lesions of scleroderma and normal skin. Immunohistochemistry staining showed that PDGF beta-receptor expression was greatly elevated in the dermis of scleroderma lesion whereas PDGF beta-receptor were expressed at low levels in normal skin. Northern blot analysis showed that cultured fibroblasts from scleroderma had higher expression of PDGF B-chain and PDGF beta-receptor mRNA than those from normal control. Two PDGF B-chain mRNA transcripts, 2.8 and 4.0 kb, were expressed. The 2.8 kb transcripts which had more efficient translation ability was the more predominantly expressed one. These results indicate that PDGF B-chain/PDGF beta-receptor signal pathway might be involved in the development of fibrosis in scleroderma, and that the 2.8 kb PDGF B-chain mRNA transcript may be the main modulation gene.

Effects of aminoglutethimide on hypothalamic-pituitary-adrenal functions.

This study was performed on four groups of subjects, including 10 patients with Cushing's disease, 10 patients with simple obesity, 8 patients with hypopituitarism and 13 normal subjects. The study was conducted by measuring the sequential changes of plasma ACTH, serum cortisol, 24-h UFC, 24-h 17 KS and 24-h 17 KGS following aminoglutethimide (AG) administration. The results suggest that normal subjects showed sequential changes of hypothalamic-pituitary-adrenal hormone concentrations with normal feedback regulation of the axis following AG administration. Patients with Cushing's disease had obvious autonomy in the production of ACTH from the pituitary. Patients with simple obesity might display abnormality to some degree in the production from the pituitary. Patients with hypopituitarism lost the capacity of ACTH production in various degrees because of pituitary lesions.