Unintrusive multi-cancer detection by circulating cell-free DNA methylation sequencing (THUNDER): development and independent validation studies.

BACKGROUND: Early detection of cancer offers the opportunity to identify candidates when curative treatments are achievable. The THUNDER study (THe UNintrusive Detection of EaRly-stage cancers, NCT04820868) aimed to evaluate the performance of enhanced linear-splinter amplification sequencing, a previously described cell-free DNA (cfDNA) methylation-based technology, in the early detection and localization of six types of cancers in the colorectum, esophagus, liver, lung, ovary, and pancreas. PATIENTS AND METHODS: A customized panel of 161 984 CpG sites was constructed and validated by public and in-house (cancer: n = 249; non-cancer: n = 288) methylome data, respectively. The cfDNA samples from 1693 participants (cancer: n = 735; non-cancer: n = 958) were retrospectively collected to train and validate two multi-cancer detection blood test (MCDBT-1/2) models for different clinical scenarios. The models were validated on a prospective and independent cohort of age-matched 1010 participants (cancer: n = 505; non-cancer: n = 505). Simulation using the cancer incidence in China was applied to infer stage shift and survival benefits to demonstrate the potential utility of the models in the real world. RESULTS: MCDBT-1 yielded a sensitivity of 69.1% (64.8%-73.3%), a specificity of 98.9% (97.6%-99.7%), and tissue origin accuracy of 83.2% (78.7%-87.1%) in the independent validation set. For early-stage (I-III) patients, the sensitivity of MCDBT-1 was 59.8% (54.4%-65.0%). In the real-world simulation, MCDBT-1 achieved a sensitivity of 70.6% in detecting the six cancers, thus decreasing late-stage incidence by 38.7%-46.4%, and increasing 5-year survival rate by 33.1%-40.4%, respectively. In parallel, MCDBT-2 was generated at a slightly low specificity of 95.1% (92.8%-96.9%) but a higher sensitivity of 75.1% (71.9%-79.8%) than MCDBT-1 for populations at relatively high risk of cancers, and also had ideal performance. CONCLUSION: In this large-scale clinical validation study, MCDBT-1/2 models showed high sensitivity, specificity, and accuracy of predicted origin in detecting six types of cancers.

[Synchronous robotic resection for colorectal liver metastasis].

Surgical resection remains the only curative therapy for colorectal adenocarcinoma and liver metastasis. Synchronous robotic resection for colorectal liver metastasis (CRLM) offers the advantage of avoiding double surgical stress, while providing the benefits of small incision, quicker recovery, shorter hospital stay and faster postoperative adjuvant therapy. Compared with the laparoscopic approach, robotic approach is mostly suitable for rectal cancer liver metastasis, which is associated with low conversion rate, good nerve protection, high success rate for major hepatectomy and resection of difficult segments. Appropriately selected patients, multidisciplinary cooperation and skillful robotic surgeons are the key to success. Current data have demonstrated the feasibility and safety of synchronous robotic resection for CRLM. With the coming randomized controlled trial data and evolution of robotic surgical system, the future of synchronous robotic resection for colorectal liver metastasis is promising.

[Experience of teaching and training for medical students at gastrointestinal surgery department under COVID-19 epidemic situation].

In hospitals and medical schools as densely populated sites with high risk of coronavirus disease 2019 (COVID-19), it is vital to adjust the teaching and training strategy for medical students to ensure curriculum completion with safety. This article aims to introduce the experience of teaching and training for medical students under the epidemic situation at Department of Surgery, Shanghai Medical College, Fudan University and Zhongshan Hospital. The content includes exploring diversified online teaching models for undergraduate surgery courses and clinical practice, carrying out online graduate education and dissertation plans, and strengthening comprehensive education of medical humanity combined with knowledge of COVID-19 prevention. Through implementation of the above teaching strategies, scheduled learning plans of medical students can be well completed in an orderly, safe and quality-ensured manner. Our experience provides practical solution of medical teaching and could be advisable for other medical colleges and teaching hospitals.

[Risk factors of anastomotic leakage after robotic surgery for low and mid rectal cancer].

Objective: To investigate the risk factors associated with anastomotic leakage after robotic surgery in mid-low rectal cancer. Methods: A retrospective case-control study method was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) pathologically confirmed rectal cancer; (3) distance <10 cm from tumor to anal margin; (4) robotic anterior rectal resection. Patients with previous history of colorectal cancer surgery, distant metastases or other malignant tumors, undergoing emergency surgery, with severe abdominal adhesions or those receiving combined organ resection were excluded. Based on the above criteria, 636 patients undergoing robotic radical sphincter-preserving surgery for mid-low rectal cancer in Zhongshan Hospital from January 2015 to December 2018 were included in this study, including 398 males (62.6%) and 238 females (37.4%) with a mean age of (61.9±11.3) years. Sixty-eight cases (10.7%) received neoadjuvant chemoradiotherapy. Amony the 636 included patients, 123(19.3%) underwent natural orifice specimen extraction surgery (NOSES) and 15 (2.3%) underwent preventive stoma. According to the cirteria developed by the International Rectal Cancer Research Group in 2010, the anastomotic leakage was classified as grade A (no requirement of intervention), B (requirement of intervention), and C (requirement of operation). Logistic regression was used to analyze the relationship between anastomotic leakage and clinicopathological factors. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results: Anastomotic leakage occurred in 38 cases (6.0%). The grading of anastomotic leakage was grade A in 13 cases (2.0%), grade B in 19 cases (3.0%), and grade C in 6 cases (0.9%). The 3-year disease-free survival rate of patients with anastomotic leakage and without anastomotic leakage was 83.5% and 83.6% respectively (P=0.862); the 3-year overall survival rate of the two group was 85.1% and 87.5% respectively (P=0.296). The results of univariate logistic regression analysis showed that male (P=0.011), longer operation time (P=0.042), distance ≤5 cm from tumor to anal margin (P=0.012), more intraoperative blood loss (P=0.048) were associated with anastomotic leakage (all P<0.05). NOSES was not associated with anastomotic leakage (P=0.704). Multivariate analysis confirmed that male (OR=3.03, 95%CI: 1.37 to 7.14, P=0.010), operation time ≥180 minutes (OR=2.04, 95%CI: 1.03 to 3.99, P=0.040), distance ≤5 cm from tumor to anal margin (OR=2.56, 95%CI:1.28 to 5.26, P=0.008) were independent risk factors for anastomotic leakage. Conclusion: Male, short distance from tumor to anal margin, and long operation time are independent risk factors for anastomotic leakage in patients undergoing robotic mid-low rectal cancer radical surgeries. These patients need to be cautiously treated during surgery.

LITAF, HHEX, and DUSP1 expression in mesenchymal stem cells from patients with psoriasis.

Psoriasis is a common chronic relapsing inflammatory skin disease, in which mesenchymal stem cells (MSCs) have been hypothesized to play an important role in abnormal localized inflammation and vascular proliferation observed in skin lesions. Previous studies have revealed abnormal gene expression patterns, DNA methylation status, and cytokine secretion of MSCs in psoriatic skin lesions, as well as some gene expression abnormalities related to inflammation and angiogenesis. We further verified the gene and protein expressions of inflammation-related lipopolysaccharide-induced tumor necrosis factor-alpha transcription factor (LITAF), dual-specificity protein phosphatase 1 (DUSP1), and angiogenesis-related hematopoietically expressed homeobox (HHEX) in MSCs derived from the skin lesions of psoriasis patients. The gene expression of LITAF, DUSP1, and HHEX in dermal MSCs was measured at the mRNA level using reverse transcription-polymerase chain reaction and the corresponding protein expression levels were analyzed by western blotting analysis. The gene and protein expression levels of LITAF, HHEX, and DUSP1 in dermal MSCs were significantly lower in psoriasis patients compared to controls. Amplification and western blotting results were consistent with our previously reported gene chip data. Our results suggest that dermal MSCs in psoriatic skin lesions may be involved in the development, progression, and regulation of localized inflammatory abnormalities by reducing the expression of LITAF, HHEX, and DUSP1, which are related to inflammation and angiogenesis.

Secreted protein acidic and rich in cysteine expression in human colorectal cancer predicts postoperative prognosis.

OBJECTIVE: Secreted protein acidic and rich in cysteine (SPARC) is an extracellular matrix glycoprotein involved in cell proliferation, migration and angiogenesis. The aim of this study was to assess its expression in colorectal cancer, see whether and how it correlates with clinicopathological features, and evaluate its potential prognostic significance. PATIENTS AND METHODS: SPARC expression was detected by microarrays containing 847 immunohistochemically stained specimens, and further correlated with the clinicopathological and prognostic data. The prognostic significance of its expression was assessed using Kaplan-Meier survival with log-rank tests. Multivariate regression utilizing Cox's proportional hazard model was used to evaluate prognostic factors. RESULTS: SPARC expression in the normal colorectal mucosa and colorectal cancer tissue was significantly different (p < 0.001). Low SPARC expression was found to be associated with poor prognosis, and it was unfavorably correlated with overall survival and disease-free survival in colorectal cancer patients. In addition, SPARC expression in surrounding mesenchymal and stromal cells, bowel wall invasion, lymph node metastasis, and distant metastasis were independent prognostic factors for overall survival and disease-free survival. CONCLUSIONS: Reduced expression of SPARC in colorectal cancer tissue is associated with poor prognosis and aggressive clinicopathological features. Therefore, SPARC expression could potentially be used as a prognostic predictor for colorectal cancer patients.

Liver cancer stem cells are selectively enriched by low-dose cisplatin

Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 μg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 μg/mL cisplatin, whereas 5 μg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.

Liver cancer stem cells are selectively enriched by low-dose cisplatin.

Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 µg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 µg/mL cisplatin, whereas 5 µg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.

Prospective analysis of quality of life in elderly patients treated with adjuvant chemotherapy for non-small-cell lung cancer.

BACKGROUND: Given the more comorbidities with a decline in physiologic reserve, it can be challenging to make appropriate treatment decisions in the elderly. PATIENTS AND METHODS: Here, we prospectively evaluated and compared the health-related quality of life (HRQOL) of patients aged ≥ 65 with aged <65 who were treated with a postoperative chemotherapy for completely resected stage Ib, II or IIIa non-small-cell lung cancer (NSCLC). Either four cycles of paclitaxel (Taxol)-carboplatin (PC) or vinorelbine-cisplatin (NP) was used. The HRQOL was assessed with EORTC QLQ-C30 and EORTC QLQ-LC13. RESULTS: Between October 2008 and October 2011, a total of 139 patients (aged <65, n = 73; ≥ 65, n = 66) were enrolled, and 127 (91.4%) completed the questionnaire. Overall, the quality of life (QOL) in elderly patients did not significantly deteriorate with adjuvant chemotherapy and the time trend of QOL in elderly patients was similar to that of younger patients. Although the elderly suffered from increased treatment-related adverse events involving sore mouth, peripheral neuropathy and alopecia compared with the baseline, the same time trends were also observed in younger group. The mean dose intensities (MDIs) for PC and NP regimen were not significantly different between the two age groups. CONCLUSIONS: Postoperative chemotherapy did not substantially reduce HRQOL in elderly NSCLC patients, and HRQOL during and after adjuvant chemotherapy did not significantly differ by age.

Cytotoxic flavonoids with isoprenoid groups from Morus mongolica.

A new pyranoflavanone, sanggenol L (1), a Diels-Alder type adduct regarded as a cycloaddition product of a dehydrogeranylflavanone and a prenylchalcone, sanggenol M (2), along with four new 2-arylbenzofurans with isoprenoid units, mulberrofurans W-Z (3-6), were isolated together with 10 known flavonoids from Chinese Morus mongolica. The structures of these novel compounds were elucidated by spectroscopic methods. All flavanones investigated here showed higher cytotoxicity against human oral tumor cell lines (HSC-2 and HSG) than against normal human gingival fibroblasts (HGF). Among them, the cytotoxicity of compound 2 and the Diels-Alder type flavanone sanggenon C (7) isolated from Morus cathayana were the most potent. On the other hand, seven 2-arylbenzofurans exhibited lower cytotoxicity and tumor specificity as compared with flavanones.

Diagnosis and management of allergic fungal sinusitis with orbital involvement.

PURPOSE: Allergic fungal sinusitis (AFS) is a noninvasive disease characterized by recurrent sinusitis. This condition is commonly treated with surgical debridement and several months of systemic corticosteroids. The treatment of AFS is examined in this study. METHODS: A retrospective case series of three patients with AFS. RESULTS: All three patients were treated with surgical debridement and less than one month of systemic corticosteroids. The patients then were treated with intranasal corticosteroids and monitored closely. Antifungal therapy was not used. All three patients remained disease-free during follow-up ranging from 12 months to 36 months. CONCLUSIONS: Surgical debridement and systemic corticosteroids for less than four weeks followed by intranasal corticosteroids may provide long-term control of AFS. Additional study is recommended to examine further the optimal treatment for AFS.

The effects of two different cisplatin-based chemotherapy regimens on advanced non-small cell lung cancer.

BACKGROUND: Many different cisplatin-based regimens have been used on advanced non-small cell lung cancer (NSCLC) in previous studies but there have been few such references in Taiwan. In this study, we evaluated the efficacy and toxicity of two different regimens including 5-Fluorouracil, Leucovorin, Etoposide and cisPlatin (FLEP) and cisPlatin, Etoposide and Mitomycin (PEM) in the treatment of patients with advanced NSCLC. METHODS: We retrospectively analyzed the records of 44 patients with NSCLC who met the selection criteria from February 1995 through April 1998. All of them were confirmed, using histologic tests, that they were in advanced stages, i.e. stage IIIB or IV. Twenty-two patients received FLEP and 22 patients received PEM. RESULTS: Three patients with FLEP therapy and 3 patients with PEM therapy had partial response. No patient had complete response. The response rate was 13.6% in both groups, respectively. The median survival was 160 +/- 30 (median + SD) days for patients with FLEP therapy and 263 +/- 104 days for patients with PEM therapy. The factors that were associated with longer survival in all patients included response (Stable Disease vs Disease Progression p = 0.004, Partial Response vs Disease Progression p = 0.047) and regimen of chemotherapy (PEM vs FLEP p = 0.008). The major clinically significant toxicity was myelosupression. CONCLUSION: The responses to regimens, FLEP and PEM, were low in our study groups that might be due to the low dose of cisplatin and etoposide in our regimens. The patients with response to chemotherapy and PEM therapy had longer median survival than those who underwent FLEP therapy.

Periocular atypical mycobacterial infections.

OBJECTIVE: To examine the clinical characteristics and management of periocular infections caused by atypical mycobacteria. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Six patients were identified with periocular atypical mycobacterial infections: four with Mycobacterium chelonae and two with Mycobacterium fortuitum. INTERVENTION: The treatment of these infections included removal of the foreign bodies with debridement of the lesions. Specimens were sent for histopathologic examination, routine cultures, and fresh tissue for culture after homogenization. MAIN OUTCOME MEASURES: A retrospective review of culture-proven atypical mycobacterial infections involving the periocular tissues was performed. Charts were reviewed for age, gender, infectious organism, medical history, surgical history, presenting symptoms, clinical features, and treatment. RESULTS: Four associations with infection were identified in these patients: immunosuppression, nasolacrimal duct obstruction, the presence of a foreign body, and a history of recent surgery. All six of the patients had at least one of these associations and five of the patients had at least two. Clinical characteristics that may distinguish atypical mycobacterial infections from acute bacterial infections include subacute presentation, firm nodular lesions, mild erythema, mild tenderness, and minimal purulent discharge. All patients had resolution of their infections after debridement and several weeks of systemic antibiotic therapy guided by susceptibility testing. CONCLUSIONS: Periocular atypical mycobacterial infections are uncommon. The clinical history and examination can raise the suspicion of this infection by revealing the clinical characteristics of these infections. Treatment includes removal of foreign bodies, debridement, and long-term systemic antibiotic therapy.

Skeletal muscle contractions stimulate cGMP formation and attenuate vascular smooth muscle myosin phosphorylation via nitric oxide.

Nitric oxide generated by neuronal nitric oxide synthase in contracting skeletal muscle fibers may regulate vascular relaxation via a cGMP-mediated pathway. Neuronal nitric oxide synthase content is greatly reduced in skeletal muscles from mdx mice. cGMP formation increased in contracting extensor digitorum longus muscles in vitro from C57 control, but not mdx mice. The increase in cGMP content was abolished with NG-nitro-L-arginine. Sodium nitroprusside treatment increased cGMP levels in muscles from both C57 and mdx mice. Skeletal muscle contractions also inhibited phenylephrine-induced phosphorylation of smooth muscle myosin regulatory light chain. Arteriolar dilation was attenuated in contracting muscles from mdx but not C57 mice. NO generated in contracting skeletal muscle may contribute to vasodilation in response to exercise.

Delay of estradiol-induced surge secretion of LH in gilts by intracerebroventricular injection of morphine.

In Experiment 1, ovariectomized (OVX) gilts, 143 d old and 58.5 +/- 1.8 kg BW, received 10 micrograms estradiol benzoate (EB)/kg BW i.m. and either 500 micrograms of the endogenous opioid peptide (EOP) agonist, morphine (MOR), in saline (SAL; n = 5), or SAL (n = 4) intracerebro-ventricularly at 40 and 48 hr after EB. With the exception of one MOR-treated gilt, which was deleted from Experiment 1, LH secretion was suppressed for at least 50 hr in all gilts. Timing of the LH surge was similar among gilts. However, total LH secreted was greater (P < 0.05) after SAL than MOR. The experiment was repeated at 179 d of age and 78.6 +/- 1.2 kg BW, except that treatments were reversed among gilts. Emergence of the LH surge was delayed (P < 0.005) by 10.8 hr and time to maximum LH concentration (P < 0.05) by 6.8 hr after MOR than after SAL. Magnitude and total LH secreted were not different among gilts. In Experiment 2, gilts which had displayed estrous cycles of 18-22 d were OVX and treated as in Experiment 1, except MOR (n = 3) or SAL (n = 4) were injected 10 hr later than in Experiment 1, i.e., at 50 and 58 hr after EB. Secretion of LH was suppressed for at least 60 hr in both groups. Time to emergence of the LH surge was delayed by 27 hr (P < 0.05) after MOR compared to after SAL. However, other parameters of the surge were not different among gilts. Thus, EOP modulate LH surge secretion negatively in the pig.

Intensive chemotherapy plus recombinant human granulocyte-colony stimulating factor support for distant metastatic nasopharyngeal carcinoma. A preliminary report.

Nasopharyngeal carcinoma (NPC) has been shown to be highly responsive to chemotherapy. The major limiting toxicity was myelotoxicity. Recently, the role of granulocyte colony-stimulating factor (G-CSF) in reducing chemotherapy-induced neutropenic sepsis has been well established. In this study, we tested whether recombinant human G-CSF (rhG-CSF) could effectively support the bone marrow function in both previously untreated and pretreated metastatic NPC patients receiving intensive chemotherapy. Twelve patients with distant metastatic disease, 5 newly diagnosed (group A) and 7 pretreated patients (group B), were enrolled to receive BEC (bleomycin, epirubicin and cisplatin), followed by rhG-CSF support (50 microg/m2 s.c. daily for 10 days) every 4 weeks for two cycles. Four patients in group A completed the treatment as scheduled while only 2 patients in group B did. After the first treatment cycle, 6 patients (50%) had grade III-IV myelosuppression. Five of the patients were from group B. The mean values of the white cell count nadir were 2,680 (range 1,200-3,700) in group A and 1,343 (range 400-2,900) in group B (p = 0.0386). Neutropenia-associated fever occurred in 7 patients, 6 of whom had received previous treatment. There were 2 deaths due to toxicity, and both patients had liver metastases within 6 months following radiation. After 24 months of follow-up, only 1 patient is still alive. Our preliminary results suggest that in previously treated metastatic NPC patients, bone marrow suppression is still the major limiting toxic side effect of aggressive chemotherapy, especially for those patients with liver recurrences within 6 months after irradiation and despite rhG-CSF support.

Opioid modulation of gonadotropin releasing hormone release from the hypothalamic preoptic area in the pig.

Two experiments (Exp) were conducted to examine in vitro the release of gonadotropin releasing hormone (GnRH) from the hypothalamus after treatment with naloxone (NAL) or morphine (MOR). In Exp 1, hypothalamic-preoptic area (HYP-POA) collected from 3 market weight gilts at sacrifice and sagittally halved were perifused for 90 min prior to a 10 min pulse of morphine (MOR; 4.5 x 10(-6) M) followed by NAL (3.1 x 10(-5) M) during the last 5 min of MOR (MOR + NAL; n = 3). The other half of the explants (n = 3) were exposed to NAL for 5 min. Fragments were exposed to KCl (60 mM) at 175 min to assess residual GnRH releasability. In Exp 2, nine gilts were ovariectomized and received either oil vehicle im (V; n = 3); 10 micrograms estradiol-17 beta/kg BW in 42 hr before sacrifice (E; n = 3); .85 mg progesterone/kg BW in twice daily for 6 d prior to sacrifice (P4; n = 3). Blood was collected to assess pituitary sensitivity to GnRH (.2 microgram/kg BW) on the day prior to sacrifice. On the day of sacrifice HYP-POA explants were collected and treated as described in Exp 1 except tissue received only NAL. In Exp 1, NAL increased (P < .05) GnRH release. This response to NAL was attenuated (P < .05) by coadministration of MOR. Cumulative GnRH release after NAL was greater (P < .05) than after MOR + NAL. All tissues responded similarly to KCl with an increase (P < .05) in GnRH release.(ABSTRACT TRUNCATED AT 250 WORDS)