Anaplastic thyroid carcinoma (ATC) is the most malignant tumor of endocrine system, which is an urgent medical problem to be solved. At present, immunotherapy studies on ATC mainly include cutting off the recruitment of tumor-associated macrophage (TAM), inducing the reprogramming of TAM and restoring its phagocytic function, targeting related immune checkpoints on T cells and natural killer cells, tumor vaccines based on oncolytic viruses and dendritic cells, and adoptive immunotherapy. Among them, immunotherapy strategies represented by targeted blocking of programmed death-1/programmed death ligand-1 at immune checkpoint have been preliminarily confirmed to benefit ATC patients, especially the combination of molecular targeted inhibitors and immunotherapy has shown excellent therapeutic effects. Due to the great heterogeneity of ATC, it is expected to provide more therapeutic strategies for patients of ATC by carrying out various immunotherapy studies including biological, immune and cellular therapies and exploring the therapeutic potential of the next generation of immune checkpoint inhibitors. This article reviews the potential immunotherapeutic targets of ATC and the progress of immunotherapy.

Anaplastic thyroid carcinoma (ATC) is a highly malignant and aggressive thyroid malignancy with rapid onset and poor prognosis. There is no effective treatment for ATC yet. Molecular targeted therapy provides a new idea for ATC treatment. Tyrosine kinase inhibitor lenvatinib has potential in treating ATC patients with favorable efficacy in clinical trials. The effectiveness of the v-raf murine sarcoma viral oncogene homologue B1 (BRAF) gene inhibitor dabrafenib in combination with trametinib for the treatment of BRAF^V600E positive ATC patients has been demonstrated in clinical trials. The NCCN clinical practice guidelines in oncology has proposed dabrafenib in combination with trametinib as the preferred modality for the treatment of patients with BRAF^V600E positive ATC. The immune checkpoint inhibitor pembrolizumab can be applied to treat thyroid cancer with high tumor mutational load and may be considered as the preferred modality for the treatment of ATC patients with high programmed death ligand-1 expression. The mammalian target of rapamycin pathway inhibitors, peroxisome proliferators-activated receptor γ agonists, endothelial growth factor receptors-targeting monoclonal antibody cetuximab and novel vascular blocker fosbretabulin are still in the clinical research stage, which are expected to provide new directions for the development of novel targeted drugs. This article reviews the current research progress on targeted drugs for the treatment of ATC.

Gasless endoscopic thyroidectomy through unilateral axillary approach has advantages of clear vision, simple manipulation, short learning curve, hidden surgical incision, no postoperative neck scar, and less swallowing discomfort. During the procedure the separation path goes through thoracic muscle surface, sternocleidomastoid gap and jugular vein, which may meet various variations of neck muscles, blood vessels and nerves. With the “three-propulsion” suspension cavity construction method the procedure advances the dissection from the axillary incision to clavicle, from the clavicle to sternocleidomastoid gap and from the sternocleidomastoid gap to thyroid. Combined with intraoperative hanging upward hook it can establish a good cavity for the subsequent surgical operation. This article introduces the main steps, key points and attentions of the “three-propulsion”suspension cavity construction method in gasless endoscopic thyroidectomy through unilateral axillary approach.

The thyroid surgery with lateral cervical incision uses the sternocleidomastoid intermuscular approach through the gap between band muscles and carotid sheath to reach the surgical field. The recurrent laryngeal nerve and upper and lower parathyroid glands are first identified, the upper pole vessels are severed; then the Berry ligament is separated; the isthmus is severed, and the thyroid gland is finally removed. This approach can avoid the trauma of the skin and muscle tissue in the anterior neck region, to relieve the pressure on the neck and swallowing stretch feeling for patients after surgery. The surgical modality is effective and less time-consuming. This article gives a detailed introduction to the standardized procedures and some key points of thyroid surgery with lateral cervical incision.

Objective: To investigate the prognostic factors of patients with persistent/recurrent differentiated thyroid carcinoma (DTC) especially with external invasive persistent recurrent DTC after comprehensive treatment. Methods: The clinical data of 525 patients with persistent/recurrent DTC who underwent surgical treatment from August 2011 to June 2021 in the Department of Head and Neck Surgery of Jiangsu Cancer Hospital were retrospectively analyzed. The prognostic factors affecting overall survival (OS) and relapse-free survival (RFS) of persistent/recurrent DTC, especially external invasive persistent/recurrent DTC were analyzed. Results: Among 525 patients, 318 patients underwent thyroidectomy, 359 patients underwent central lymph node dissection, and 409 patients underwent lateral cervical lymph node dissection. Among 493 followed-up patients, 5-year OS and RFS were 95.10% and 89.60%, 8-year OS and RFS were 91.80% and 81.30%. Cox regression analysis showed that in patients with persistent/recurrent DTC after comprehensive treatment, age ≥55?years at reoperation after recurrence, male gender and distant metastasis were independent risk factors of OS (all P<0.05); while the simultaneous invasion of thyroid and lymph nodes, multiple organ invasion and the number of previous operations ≥2 were independent risk factors of RFS (allP<0.05). In patients with external invasive persistent/recurrent DTC after comprehensive treatment, age ≥55?years at reoperation after recurrence and male gender were independent risk factors of OS (bothP<0.05); while multiple organ invasion and the number of previous operations ≥2 were independent risk factors of RFS (bothP<0.05).Conclusions: Male patients aged 55?years old and above, with distant metastasis have a higher risk of poorer prognosis in persistent/recurrent DTC; while patients with simultaneous external invasion of thyroid and lymph nodes, multiple organ invasion and the number of previous operations ≥2 are more likely to relapse. For external invasive persistent/recurrent DTC, male patients aged 55?years old and above have a higher risk of poorer prognosis; while patients with multiple organ invasion and the number of previous operations ≥2 are more likely to have recurrence.

Objective: To evaluate the application of endoscopic thyroidectomy using gasless axillary approach (ET-GA) for low-risk papillary thyroid carcinoma (PTC). Methods: Patients with T1N0M0 Ⅰ PTC undergoing unilateral thyroid lobectomy with central neck dissection in Taizhou Cancer Hospital during January 2019 to June 2021 were enrolled in the study, including 35 cases treated with ET-GA (ET-GA group) and 35 cases treated with conventional open thyroidectomy (COT group). The surgical treatment effect, cosmetic effect and the effect on neck function were compared between two groups. Neck function was evaluated by neck pain score, neck injury index and dysphagia index. Cosmetic effect was evaluated by cosmetic effect satisfaction score. Results: In the ET-GA group, all unilateral thyroid lobectomy with central neck dissection were successfully completed, and no case was converted to open surgery. The number of central lymph nodes dissected in the ET-GA was not statistically different from that in the COT group (P>0.05), but the operation time was longer than that of the COT group (P<0.01). In the ET-GA group, 2 cases (5.7%) had transient vocal cord paralysis and 1 case (2.9%) had postoperative bleeding. In the COT group, 1 case (2.9%) had transient vocal cord paralysis, no postoperative bleeding. There was no significant difference in the complication rate between two groups (P>0.05). At 3?d and 3?months postoperatively, there was no significant difference in neck pain score and neck injury index between two groups (bothP>0.05); dysphagia index was lower in ET-GA group, but the difference was not statistically significant (P>0.05). The cosmetic effect satisfaction score of ET-GA group was higher than that in the COT group at 3?months postoperatively (4.3±0.6 vs.3.2±1.0,P<0.01).Conclusion: ET-GA has the same efficacy and safety as conventional open thyroidectomy in the treatment of low-risk PTC, and it improves the satisfaction of postoperative cosmetics.

Objective:To establish a risk probability model for residual metastatic lymph nodes in patients with papillary thyroid microcarcinoma (PTMC) after cervical central lymph node dissection (CLND). Methods:The clinical data of patients with PTMC treated in the First Affiliated Hospital of Kunming Medical University from 2007 to 2020 were retrospectively reviewed. All patients underwent thyroidectomy with CLND, and at least one lymph node was examined. Based on the distribution characteristics of metastatic lymph nodes from this retrospective cohort, a probabilistic model for the risk of residual metastatic lymph node was established. β-Binomial distribution was used to estimate the probability of residual metastatic lymph node as a function of the number of lymph nodes examined. Results: Among 5399 patients included in the probabilistic model, central lymph node metastases were observed in 1664 cases (30.8%). After model correction, the real lymph node metastasis rate increased from 30.8% to 38.9%. The probability of false negative of central lymph node was estimated to be 31.3% for patients with a single node examined, while decreased to 10.0% and 4.9% when 7 and 12 nodes were examined, respectively. In the sensitivity analysis limited to patients with or without Hashimoto thyroiditis, the performance of probability model was also satisfactory. Conclusions: The established risk probability model in this study quantifies the risk of residual metastatic lymph nodes after CLND in patients with PTMC, which can be used as complementary indicators for the risk of recurrence/persistence disease at postoperative evaluation. The study also provides a new method to evaluate the impact of residual metastatic lymph nodes on the prognosis of tumor patients through retrospective data.

Objective: To investigate risk factors of lateral cervical lymph node metastasis (LLNM) in patients with medullary thyroid carcinoma (MTC). Methods: Published studies regarding clinicopathological factors of LLNM in MTC were searched in PubMed, Web of Science, Embase, Cochrane library, Wanfang date and CNKI. Statistical analysis was performed using Stata 14.0 software. The mean and standard deviation from the sample size, range, median, and interquartile range was estimated. Odds ratio (OR) or standard mean difference (SMD) with 95% confidence interval (CI) of related factors were analyzed by fixed/random-effects models. Egger’s test and Begg’s test were applied to assess the publication bias of the literature. This study was registered with PROSPERO (CRD42021254955). Results: Fifteen studies involving 1424 patients were included in the analysis, among whom 543 cases had LLNM (38.13%). Meta-analysis revealed that an increased risk of LLNM was associated with male gender (OR=1.64, 95%CI: 1.29–2.09, Z=4.06, P<0.01), tumor diameter≥1cm (OR=5.09, 95%CI: 2.43–10.67, Z=4.31, P<0.01), multifocality (OR=2.55, 95%CI: 1.79–3.61, Z=5.22, P<0.01), capsule invasion (OR=7.80, 95%CI: 4.84–12.55, Z=8.46, P<0.01), extracapsular extension (OR=9.46, 95%CI: 5.66–15.81, Z=8.58, P<0.01), cervical central lymph node metastasis (OR=23.58, 95%CI: 9.44–58.87, Z=6.77, P<0.01), elevated preoperative calcitonin (SMD=1.17,95%CI: 0.67–1.67, Z=4.56, P<0.01), spiculated margin on ultrasonography (OR=4.32, 95%CI: 2.43–7.68, Z=4.99, P<0.01), irregular shape on ultrasonography (OR=6.81, 95%CI: 3.64–12.73, Z=6.01, P<0.01); while age ≥ 45 years (OR=1.22, 95%CI: 0.65–2.29, Z=0.62, P>0.05), elevated preoperative carcinoembryonic antigen (SMD=0.95, 95%CI: –0.48–2.38, Z=1.30, P>0.05) and calcification on ultrasonography (OR=1.28, 95%CI: 0.75–2.18, Z=0.92, P>0.05) were not associated with LLNM.Conclusion: Male gender, tumor diameter≥1?cm, multifocality, capsule invasion, extracapsular extension, central lymph node metastasis, elevated preoperative calcitonin, spiculated margin and irregular shape on ultrasonography are risk factors for LLNM in MTC, when these clinical and ultrasonic features are present, lateral neck lymph node dissection is recommended.

Objective: To explore the association between napping status and depressive symptoms in urban residents during the coronavirus disease 2019 (COVID-19) epidemic. Methods: The survey was embedded in the Wellness Living Laboratory-China (WELL China) cohort study. Health and lifestyle information during the COVID-19 epidemic were obtained via the telephone interview from April 8, 2020 to May 29, 2020. A total of 3075 residents aged 18 to 80?years from Gongshu district of Hangzhou city with complete data were included in the analyses. The World Health Organization-Five Well-being Index (WHO-5) was used to measure depressive symptoms. Multiple logistic regression model was used to assess the association between napping status and depressive symptoms in the participants. Results: The prevalence of depressive symptoms was 20.6% in the participants during the COVID-19 epidemic. Daytime napping behavior, especially napping time ≤30?min, was associated with a lower risk of prevalent depressive symptoms (OR=0.61, 95%CI: 0.47–0.79, P<0.01) and incident depressive symptoms in the population (OR=0.66, 95%CI: 0.50–0.88, P<0.01). Among those with depressive symptoms at baseline, napping time ≤ 30?min was beneficial for the outcome of depressive symptoms (OR=0.42, 95%CI: 0.21–0.82, P<0.05).Conclusion: One in five urban residents have depressive symptoms during the COVID-19 epidemic, and a short nap during the day may be a protective factor against depressive symptoms.

Objective:To explore the application value of nanopore sequencing technique in the diagnosis and treatment of secondary infections in patients with severe coronavirus disease 2019 (COVID-19). Methods:A total of 77 clinical specimens from 3 patients with severe COVID-19 were collected. After heat inactivation, all samples were subjected to total nucleic acid extraction based on magnetic bead enrichment. The extracted DNA was used for DNA library construction, then nanopore real-time sequencing detection was performed. The sequencing data were subjected to Centrifuge software database species matching and R program differential analysis to obtain potential pathogen identification. Nanopore sequencing results were compared with respiratory pathogen qPCR panel screening and conventional microbiological testing results to verify the effectiveness of nanopore sequencing detection. Results: Nanopore sequencing results showed that positive pathogen were obtained in 44 specimens (57.1%). The potential pathogens identified by nanopore sequencing included Klebsiella pneumoniae, Staphylococcus epidermidis, Enterococcus faecium and Dolosigranulum pigrum, et al. Klebsiella pneumoniae, Staphylococcus epidermidis, Enterococcus faecium were also detected in clinical microbiological culture-based detection; Klebsiella pneumoniae was detected in respiratory pathogen screening qPCR panel; Dolosigranulum pigrum was only detected by the nanopore sequencing technique. Comprehensive considerations with the clinical symptoms, the patient was treated with antibiotics against Dolosigranulum pigrum, and the infection was controlled. Conclusion:Nanopore sequencing may assist the diagnosis and treatment of severe COVID-19 patients through rapid identification of potential pathogens.

Objective: To investigate the expression of low-density lipoprotein receptor-related protein 6 (LRP6)/β-catenin pathway related proteins in adipose tissue of rats with intrauterine growth restriction with catch-up growth (CG-IUGR). Methods: SD rats were randomly divided into nutrition-restriction rats and normal feed rats during pregnancy. CG-IUGR model was established by reducing the number of offspring in the nutrition-restriction rats (CG-IUGR group); while the rats in the control group were offspring of the normal feed pregnant rats. In order to exclude the interference of gender, male offspring mice were selected in both the CG-IUGR group and the control group in the following studies. The CG-IUGR group and the control group were subjected to glucose tolerance test at 12 weeks of age, and the perirenal adipose tissue samples were taken to observe the adipose structure by HE staining. Expression of LRP6, β-catenin and insulin receptor substrate 1 (IRS-1) in adipocytes were examined by confocal microscopy. Protein expression of LRP6, β-catenin and IRS-1 were measured by Western blotting. Results: Blood glucose level and the area under the cure of CG-IUGR group were significantly higher than that of control group (both P<0.05). Adipocyte size in the CG-IUGR group was significantly larger than that of control group, and the expression of LRP6, β-catenin and IRS-1 protein in adipose tissue of the CG-IUGR group was significantly lower than that of control group (allP<0.05).Conclusion: The expression of LRP6/β-catenin pathway related proteins is reduced in the adipose tissue in CG-IUGR rats, probably contributing to the insulin resistance in these rats.

Objective:To investigate the effects of θ–γ neural oscillation stimulation in hippocampal CA3 area on spatial cognition ability in rats. Methods:According to the results of Y maze shock avoidance training, the rats were divided into fast avoidance response group and general avoidance response group. Using endogenous θ–γ neural oscillations from the fast avoidance response rats to perform deep brain stimulation in vivo to the left and right hippocampal CA3 region of rats with general avoidance response, then the spatial cognition was tested by Y maze shock avoidance training. The variation of θ oscillation and low-γ neural oscillation phase-amplitude coupling (PAC) in CA3 area was analyzed by wavelet packet extraction technique. Western blotting was used to detect the expression of N-methyl-D-aspartate receptor 2B subunit (NR2B) and postsynaptic density(PSD)-95 in hippocampal tissues of rats to explore its molecular mechanism. Results: Compared with the general avoidance response rats, the days to reach the standard, the training number, the correct response time and the error reaction number in simulated stimulus avoidance response rats were significantly reduced, but the correct response rate was significantly increased (all P<0.01); the θ–γ neural oscillations PAC in the hippocampal CA3 region in the simulated stimulus avoidance response rats (3–5?Hz and 30–34, 38–42, 44–48?Hz; 5–7?Hz and 42–46, 44–48, 54–58?Hz) were significantly higher than that in the general avoidance response rats (allP<0.05). Meanwhile, the protein expressions of NR2B and PSD-95 in hippocampal tissues were significantly increased (bothP<0.05) in simulated stimulus avoidance response rats.Conclusion: The spatial cognition of normal avoidance response rats can be significantly improved by endogenous θ–γ neural oscillation stimulation to hippocampal CA3 region, which may be caused by the enhancement of synaptic plasticity mediated by NR2B and PSD-95.

Objective: To investigate the feasibility, methods and efficacy of ultrasound-guided nitinol stent implantation for the treatment of early recurrent stenosis of arteriovenous fistula (AVF). Methods: Thirty patients with early recurrent stenosis after percutaneous transluminal angioplasty (PTA) who received ultrasound-guided nitinol stent implantation in Sir Run Run Shaw Hospital of Zhejiang University from April 2018 to July 2020 were followed up. The imaging features of the procedure and the interventional devices were observed under ultrasonography. The technical success rate and the clinical success rate as well as the incidence of complication were assessed. The post-interventional primary patency rates of access circuit, primary patency rates of target lesion and secondary patency rates were estimated. Results: Ultrasonography was able to demonstrate the operation process and the interventional devices clearly. The technical and clinical success rates were both 100.0%. Eight patients had in-stent restenosis, which were treated by PTA. The post-interventional primary patency rates of the access circuit after 3, 6, 9 and 12?months were 91.3%, 86.2%, 86.2% and 64.2%, respectively; the post-interventional primary patency rates of target lesion were 100.0%, 100.0%, 86.4% and 69.3%, respectively; the post-interventional secondary patency rates were 100.0%, 100.0%, 100.0% and 94.4%, respectively. Compared with previous PTA in these cases, stent implantation had a higher post-interventional primary patency rates of target lesion and a lower cost-effectiveness (both P<0.05). No other complications such as vascular rupture, pseudohemangioma, stent infection, stent displacement and stent exposure were observed during the follow-up.Conclusion: Ultrasonography can accurately guide the nitinol stent implantation in AVF, and the technique is feasible in treatment for the early recurrent stenosis after PTA with good short- and medium-term efficacy.

As a novel genetic marker, microhaplotype can be applied in the field of forensic genetics. Microhaplotype has the advantages of high polymorphism, low mutation rate, no stutter products and short amplification fragments. Microhaplotype can effectively detect mixture, and quantitatively analyze the contributors of mixture. DNA with severe fragmentation can be successfully genotyped by microhaplotype. It can be used as ancestry informative marker to effectively divide the global continental population according to genetic structure. Microhaplotype system can provide more information than traditional short tandem repeat and help to identify complex relationships. It can provide new ideas for tumor source identification, cell line identification and prenatal paternity testing. Here we review the applications of microhaplotype, intending to provide references for forensic practice.

Primary Sj?gren syndrome is an autoimmune disease, in which a large number of lymphocytes infiltrate the exocrine glands and cause gland dysfunction. Its pathogenesis is related to the chronic inflammation of the exocrine glands caused by genetic factors, immunodeficiency or viral infection. Long-term inflammation leads to accelerated apoptosis of epithelial cells, disordered gland structure, increased expression of proinflammatory cytokine such as CXC subfamily ligand (CXCL) 12, CXCL13, B cell-activating factor (BAF), interleukin (IL)-6, interferon (IFN)-γ and tumor necrosis factor (TNF)-α in submandibular gland. With the action of antigen-presenting cells such as dendritic cells and macrophages, lymphocytes (mainly B cells) are induced to mature in secondary lymphoid organs and migrate to the submandibular gland to promotes the formation of germinal centers and the synthesis of autoantibodies. Meanwhile, innate lymphocytes, vascular endothelial cells and mucosa-associated constant T cells as important immune cells, also participated in the inflammatory response of the submandibular gland in primary Sj?gren syndrome through different mechanisms. This process involves the activation of multiple signal pathways such as JAK/STAT, MAPK/ERK, PI3K/AKT/mTOR, PD-1/PD-L1, TLR/MyD88/NF-κB, BAF/BAF-R and IFN. These signaling pathways interact with each other and are intricately complex, causing lymphocytes to continuously activate and invade the submandibular glands. This article reviews the latest literature to clarify the mechanism of submandibular gland inflammation in primary Sj?gren syndrome, and to provide insights for further research.

A child with methylmalonic acidemia and homocysteinemia cblX type presented focal seizures and epileptic spasms in early infancy, but the tandem mass spectrometry tests showed negative results during neonatal screening or acute attack. Despite treated with a variety of antiepileptic drugs, the child died at age of 4?months. The blood spot sample of the patient was retrospectively tested with ultrahigh performance liquid chromatography-tandem mass spectrometry, and the increased levels of methylmalonic acid and homocysteine were revealed. Whole exome sequencing showed that the proband had a c.202C>G(p.Q68E) hemizygous mutation inHCFC1 gene, which was inherited from his mother.