Objective:To report the long-term outcomes of Chinese rectal cancer patients after adopting a Watch and Wait (W&W) strategy following neoadjuvant therapy (NAT).Methods:This multicenter, cross-sectional study was based on real-world data. The study cohort comprised rectal cancer patients who had achieved complete or near complete clinical responses (cCRs, near-cCRs) after NAT and were thereafter managed by a W&W approach, as well as a few patients who had achieved good responses after NAT and had then undergone local excision for confirmation of pathological complete response. All participants had been followed up for ≥2 years. Patients with distant metastases at baseline or who opted for observation while living with the tumor were excluded. Data of eligible patients were retrospectively collected from the Chinese Wait-and-Watch Data Collaboration Group database. These included baseline characteristics, type of NAT, pre-treatment imaging results, evaluation of post-NAT efficacy, salvage measures, and treatment outcomes. We herein report the long-term outcomes of Chinese rectal cancer patients after NAT and W&W and the differences between the cCR and near-cCR groups.Results:Clinical data of 318 rectal cancer patients who had undergone W&W for over 2 years and been followed up were collected from eight medical centers (Peking University Cancer Hospital, Fudan University Shanghai Cancer Center, Sun Yat-sen University Cancer Center, Shanghai Changhai Hospital, Peking Union Medical College Hospital, Liaoning Cancer Hospital, the First Hospital of Jilin University, and Yunnan Cancer Hospital.) The participants comprised 221 men (69.4%) and 107 women (30.6%) of median age 60 (26-86) years. The median distance between tumor and anal verge was 3.4 (0-10.4) cm. Of these patients, 291 and 27 had achieved cCR or near-cCR, respectively, after NAT. The median duration of follow-up was 48.4 (10.2-110.3) months. The 5-year cumulative overall survival rate was 92.4% (95%CI: 86.8%-95.7%), 5-year cumulative disease-specific survival (CSS) rate 96.6% (95%CI: 92.2%-98.5%), 5-year cumulative organ-preserving disease-free survival rate 86.6% (95%CI: 81.0%-90.7%), and 5-year organ preservation rate 85.3% (95%CI: 80.3%-89.1%). The overall 5-year local recurrence and distant metastasis rates were 18.5% (95%CI: 14.9%-20.8%) and 8.2% (95%CI: 5.4%-12.5%), respectively. Most local recurrences (82.1%, 46/56) occurred within 2 years, and 91.0% (51/56) occurred within 3 years, the median time to recurrence being 11.7 (2.5-66.6) months. Most (91.1%, 51/56) local recurrences occurred within the intestinal lumen. Distant metastases developed in 23 patients; 60.9% (14/23) occurred within 2 years and 73.9% (17/23) within 3 years, the median time to distant metastasis being 21.9 (2.6-90.3) months. Common sites included lung (15/23, 65.2%), liver (6/23, 26.1%), and bone (7/23, 30.4%) The metastases involved single organs in 17 patients and multiple organs in six. There were no significant differences in overall, cumulative disease-specific, or organ-preserving disease-free survival or rate of metastases between the two groups (all P>0.05). The 5-year local recurrence rate was higher in the near-cCR than in the cCR group (41.6% vs. 16.4%, P<0.01), with a lower organ preservation rate (69.2% vs. 88.0%, P<0.001). The success rates of salvage after local recurrence and distant metastasis were 82.1% (46/56) and 13.0% (3/23), respectively. Conclusion:Rectal cancer patients who achieve cCR or near-cCR after NAT and undergo W&W have favorable oncological outcomes and a high rate of organ preservation. Local recurrence and distant metastasis during W&W follow certain patterns, with a relatively high salvage rate for local recurrence. Our findings highlight the importance of close follow-up and timely intervention during the W&W process.

Objective:To explore the relationship of anxiety,perceived stress and forbearance in the college students.Methods:Totally 3 056 college students(1 102 males and 1 954 females)in Guangdong Province were re-cruited and assessed with the Generalized Anxiety Disorder 7-item Scale(GAD-7 score of ≥10 indicates positive anxiety symptoms),Perceived Stress Scale(PSS-10),and Forbearance Scale(FS).SPSS Process model 2 was used to test the moderating effect of different types of forbearance.Results:Totally 486(15.9％)college students had anxiety.The PSS-10 scores were positively correlated with the GAD-7 scores(β=0.63),and FS scores of repres-sive forbearance and active forbearance played a moderating role between the PSS-10 scores and the GAD-7 scores respectively(β=0.05,-0.04).Conclusion:It suggests that anxiety and perceived stress are correlated with for-bearance in college students,and repressive forbearance and active forbearance may moderate the relationship be-tween perceived stress and anxiety.

Objective:To report the long-term outcomes of Chinese rectal cancer patients after adopting a Watch and Wait (W&W) strategy following neoadjuvant therapy (NAT).Methods:This multicenter, cross-sectional study was based on real-world data. The study cohort comprised rectal cancer patients who had achieved complete or near complete clinical responses (cCRs, near-cCRs) after NAT and were thereafter managed by a W&W approach, as well as a few patients who had achieved good responses after NAT and had then undergone local excision for confirmation of pathological complete response. All participants had been followed up for ≥2 years. Patients with distant metastases at baseline or who opted for observation while living with the tumor were excluded. Data of eligible patients were retrospectively collected from the Chinese Wait-and-Watch Data Collaboration Group database. These included baseline characteristics, type of NAT, pre-treatment imaging results, evaluation of post-NAT efficacy, salvage measures, and treatment outcomes. We herein report the long-term outcomes of Chinese rectal cancer patients after NAT and W&W and the differences between the cCR and near-cCR groups.Results:Clinical data of 318 rectal cancer patients who had undergone W&W for over 2 years and been followed up were collected from eight medical centers (Peking University Cancer Hospital, Fudan University Shanghai Cancer Center, Sun Yat-sen University Cancer Center, Shanghai Changhai Hospital, Peking Union Medical College Hospital, Liaoning Cancer Hospital, the First Hospital of Jilin University, and Yunnan Cancer Hospital.) The participants comprised 221 men (69.4%) and 107 women (30.6%) of median age 60 (26-86) years. The median distance between tumor and anal verge was 3.4 (0-10.4) cm. Of these patients, 291 and 27 had achieved cCR or near-cCR, respectively, after NAT. The median duration of follow-up was 48.4 (10.2-110.3) months. The 5-year cumulative overall survival rate was 92.4% (95%CI: 86.8%-95.7%), 5-year cumulative disease-specific survival (CSS) rate 96.6% (95%CI: 92.2%-98.5%), 5-year cumulative organ-preserving disease-free survival rate 86.6% (95%CI: 81.0%-90.7%), and 5-year organ preservation rate 85.3% (95%CI: 80.3%-89.1%). The overall 5-year local recurrence and distant metastasis rates were 18.5% (95%CI: 14.9%-20.8%) and 8.2% (95%CI: 5.4%-12.5%), respectively. Most local recurrences (82.1%, 46/56) occurred within 2 years, and 91.0% (51/56) occurred within 3 years, the median time to recurrence being 11.7 (2.5-66.6) months. Most (91.1%, 51/56) local recurrences occurred within the intestinal lumen. Distant metastases developed in 23 patients; 60.9% (14/23) occurred within 2 years and 73.9% (17/23) within 3 years, the median time to distant metastasis being 21.9 (2.6-90.3) months. Common sites included lung (15/23, 65.2%), liver (6/23, 26.1%), and bone (7/23, 30.4%) The metastases involved single organs in 17 patients and multiple organs in six. There were no significant differences in overall, cumulative disease-specific, or organ-preserving disease-free survival or rate of metastases between the two groups (all P>0.05). The 5-year local recurrence rate was higher in the near-cCR than in the cCR group (41.6% vs. 16.4%, P<0.01), with a lower organ preservation rate (69.2% vs. 88.0%, P<0.001). The success rates of salvage after local recurrence and distant metastasis were 82.1% (46/56) and 13.0% (3/23), respectively. Conclusion:Rectal cancer patients who achieve cCR or near-cCR after NAT and undergo W&W have favorable oncological outcomes and a high rate of organ preservation. Local recurrence and distant metastasis during W&W follow certain patterns, with a relatively high salvage rate for local recurrence. Our findings highlight the importance of close follow-up and timely intervention during the W&W process.

Objective:To explore the appropriate time of intravenous immunoglobulin(IVIG)in standard initial treatment of Kawasaki disease in children.Methods:To consult the data of children with Kawasaki disease admitted to Department of Pediatrics,Sanming First Hospital Affiliated to Fujian Medical University from January 2016 to March 2021.A total of 163 eligible cases were re-trieved during the study period,of which 53 cases in early group were treated with initial IVIG 1～4 days after onset;initial IVIG treat-ment time was 5～7 days after onset in medium-term group,with a total of 54 cases;initial IVIG treatment time was 8～10 days after on-set in late group,with a total of 56 cases.Immune function,biological markers,echocardiography,initial IVIG non-response,hospi-talization days and main clinical manifestations after treatment were compared.Results:After treatment,levels of lymphocyte subsets in three groups were improved,among which the levels of CD4+T,CD4+T/CD8+T and CD19+T were decreased,while levels of CD3+T,CD8+T and NK were increased.Levels of biological markers were lower than those before treatment;there were significant differences in levels of lymphocyte subsets,biological markers,incidence of IVIG inactive Kawasaki disease,incidence of coronary artery lesion and coronary aneurysm among the three groups after treatment(P<0.05).The differences between the early group and the medium-term group and the late group were statistically significant(P<0.05).There was no significant difference between the medium-term group and the late group(P>0.05);and the early group had the best major clinical manifestations and the shortest hospital stay.Conclu-sion:In standard initial treatment of Kawasaki disease in children,the use of IVIG within 5 days after onset of immune function recov-ery is the best,reduce level of inflammation and protein markers is the best,but also can quickly improve clinical symptoms,shorten the length of hospital stay,and has the lowest incidence of IVIG no-response to Kawasaki disease,minimal coronary artery disease and coronary aneurysm complications,may be suitable for use.

Objective:To investigate the effect of transforming growth factor (TGF- β) signal in muscle fiber itself during inflammation/immunity response on intramuscular inflammation. Methods:Sixteen wild C57BL/6 mice (wild group) and sixteen mice with skeletal muscle-specific deficiency of T βRⅡ (knock-out group) between 4-8 weeks of age were selected for this study. Acute muscle injury in mice was induced by injection of myotoxin cardiotoxin (CTX) into gastrocnemius. The differences in intramuscular inflammation were compared between the wild and knock-out groups on 0, 4, 7 and 10 d after CTX injection by observing exudation of mononuclear phagocytes, macrophages, M1 type macrophages, CD4 +T cells and helpers T cells (Th1, 2&17). Two newborn C57BL/6 wild mice and 2 SM TGF- βr2-/- knock-out mice were selected to culture primary myoblasts in vitro which were divided into 2 groups: an interferon group subjected to interferon simulation and a control group subjected to addition of an equal amount of solvent. The differences in expression of IL-6, IL-10, MCP-1, MIP-1α, H-2K b, H2-Ea, Toll-like receptor (TLR)3 and TLR7 were compared between the interferon and control groups, as well as between the wild and knock-out groups. Results:On 4&7 d after CTX injection, the ratios of mononuclear/macrophage (75.73%±3.62%, 45.27%± 2.32%), macrophages (38.67%±2.76%, 24.87%±2.19%), M1 macrophages (43.21%±0.11%, 30.43%±2.19%), CD4 +T cells (20.13%±1.62%, 5.67%±0.32%) in the muscle tissue from the knock-out mice were significantly higher than those from the wild mice (58.52%±2.43%, 29.21%±2.45%; 20.63%±2.32%, 16.23%±1.25%; 24.98%±0.35%, 14.23%±1.69%; 10.70%±0.43%, 2.50%±0.45%), with a majority of Th1&Th17 ( P<0.05). In vitro results showed that the levels of IL-6, MCP-1, MIP-1α, H-2K b, H2-Ea and TLR3 were significantly upregulated in the interferon group compared with the control group and that such upregulation in the nock-out mice was more significant than in the wild mice ( P<0.05). Conclusions:Endogenous TGF- β signal activation plays a role in the functional recovery after muscle trauma, because it is involved in the regulation of immune behavior of muscle fibers, thus affecting intramuscular inflammation and muscle regeneration.

Objective To explore the effect of exercise on vascular smooth muscle cell(VSMC) phenotype switching in hypertensive arteries and to figure out the regulatory mechanisms of mircroRNA (miR)-143/145 on Akt signaling during the process.Methods Three-month old(Wistar Kyoto rats) WKY and (spontaneously hypertensive rats) SHR were divided into 4 groups:WKY-C,SHR-C,WKY-E,and SHR-E,which were subjected to 8wk moderate treadmill training (E) or sedentary as control (C) with blood pressure being monitored.After the last bout of exercise,mesenteric arteries were isolated to determine VSMC phenotypic marker,miR143/145 expression and Akt phosphorylation.In transfection experiment in vitro,miR-145 mimic and miR-145 inhibitor were transfected into cultured VSMC,and given immunofluorescent staining using α-actin to detect the cell morphology.VSMC phenotype marker,Akt phosphorylation,and mRNA expressions of the insulin-like growth factor Ⅰ receptor (IGF-IR),Insulin receptor substrate 1 antibody(IRS-1),and p70S6K were determined.Results The blood pressure of SHR-E reduced significantly compared with that of SHR-C(P＜0.05),and the arterial thicknessof SHR-E decreased significantly (P＜0.05).The VSMC contractile marker calponin in SHR-E increased significantly when compared with SHR-C(P＜0.05),while the proliferative marker osteoppontin (OPN) in SHR-E reduced significantly than that in SHR-C(P＜0.05).The miR-145 of SHR-E was significantly enhanced(P＜0.05),while there was no significant difference in the miR-143.The Akt of SHR-E was activated more significantly than SHR-C(P＜0.05).The miR-145 overexpression by transfecting miR-145 mimic into VSMC increased α-actin significantly(P＜0.05),while miR-145 inhibitor made α-actin a decrease.Akt activation was significantly inhibited by miR-145 mimic and enhanced by miR-145 inhibitor(P＜0.05).The miR-145 significantly inhibited IRS-1 and IGF-1R mRNA(P＜ 0.05),but the targeting effects were not significant on p70S6K mRNA.Conclusions Exercise ameliorates the high blood pressure and remodels arterioles,which may rely on its regulatory role on VSMC switching from proliferative to contractile phenotype,and miR-145 is involved in this process.However,the Akt activation is not caused by the overexpression of miR-145,but through other means to promote the above VSMC switching.

Objective To study the clinical efficacy and follow-up study of ketogenic diet adding treatment for refractor epilepsy in children.Methods Cluster sampling method was employed to select children in children's hospital from January 2015 to June 2017,a total of 25 cases were diagnosed refractor epilepsy and adding ketogenic diet.Engel grade was used to evaluate the efficiency,the side effects,electroencephalogram (EEG) changes and intellectual development at 3 months,3-6 months,and more than 6 months.Results The effective rate of epileptic seizure control was 0,66.7％ and 87.5％ at 3 months,3 -6 months and ＞ 6 months respectively.The improvement rate of EEG discharge index was 33.3％,50％ and 81.3％ respectively.The improvement of intelligence development was 33.3％,50％ and 68.8％ respectively.Gastrointestinal disturbances were the main side effects.Severe side effect occurred in two cases--they had severe food refusal and were stopped the ketogenic diet adding treament.Conclusions The ketogenic diet is effective,safe,few side effects and tolerable in infants and children with refractory epilepsy.The ketogenic diet may improve cognition and behavior in addition to reducing seizure frequency,the interical epileptiform discharges (IED) index and improve the quality of life of epileptic children.However,the acceptance of ketogenic diet therapy for children is not satisfactory.The sample size is small and needs further promotion.While large samples and long-term observations are still desired to better recipes,and to provide possibly effective altemative to other therapies for refractor epilepsy.

Objective CRISPR/Cas9 genome-editing technique provides an novel method for whole genome editing in eukaryotic cells.Recently,we found that gene subtype library with smaller size and focused pur-pose is more economical and practical. In this study,we aimed to target kinases,a group of pivotal cell signal transducers,to construct a kinase knock-out library using CRISPR/Cas9 technique.The construction strategy wll al-so be discussed. Methods 10 sgRNA was designed for each kinase target.After oligo pool synthesis by semicon-ductor chip,the oligos were eluted from the chip. The oligo templates were amplified and cloned into Cas9 vector and transformed into Stble3 competent cells.Monoclonal colonies were selected for DNA sequencing. Results(1) GO analysis of 507 cell kinases showed that the cell kinases took part in a wide range of cell signaling.(2)The sgRNA pool with about 140 bp in length was successfully amplified by using oligo pool as the template and univer-sal PCR primers.(3)In 40 identified library clones,34 clones were sequenced successfully. Among them,the DNA sequencing results of 25 samples were completely consistent with the designed target sequences.But there are some mutations in the primers of 9 samples.Failure in bacteria shaking,DNA sequencing and other factors were ex-isted in the other clones. Conclusion The CRISPR/Cas9 kinase knock-out library can be widely used for screen-ing the important kinases which may mediate cell proliferation,metastasis,drug resistance and autophagy.This li-brary will play an important role in clarifying the development of disease associated with kinases.

Objective To analyze the constituent of the 32 kD protein band and its expression in schizophrenia se?rum. Methods Sixty schizophrenia patients and 58 health controls were recruited. The serum samples were collected and precipitated with 7%PEG. The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to ob?tain the abnormal 32 kD proteins band in patients. This protein band was cut and then analyzed using mass spectrometric technique. Results The 32 kD protein band was present in 38 schizophrenia patients but not in control and positive rate was 63.33%. The mass spectrometric analysis showed that 32 kD protein band contained 14 proteins ranging from 30 kD to 35 kD, including 6 high-frequency proteins (cDNA coded protein 1 and 2, Apolin protein A-1, Isoform 2 of ficolin-2, Complement factor H and clusterin) and 8 low-frequency proteins (IgG H chain, zinc-alphg-2-glycoprotein, fermitin,family apolin protein L-1, isoform 10 of collectin-1, purine nucleoside, anne xin and cDNA coded protein 3). Three cD?NA coded unknown proteins were highly similar to complement C4-B, β2-glycoprotein and erythrocyte band 7 integral membrane protein. Conclusion There is a unknown specific 32 kD protein that is consisted mainly of fourteen proteins in serum of schizophrenia.

Objective To detect the disparity of three biological molecules Caveolin-1,IL-1β,VEGF in cerebrospinal fluid of children with viral encephalitis at the different stages; to explore the role of Caveolin-1,IL-1β,VEGF in the pathogenesis of viral encephalitis;and to evaluate their clinical significance in assessing the severity and prognosis of viral encephalitis.Methods We recruited 65 inpatients children with viral encephalitis in the Second Neurology Department of Hunan Children's Hospital from July 2011 to July 2012.Subjects were divided into 2 groups:54 cases of acute phase and 11 cases of recovery phase.According to the clinical manifestations,they were re-divided into 40 patients with mild viral encephalitis and 25 cases of severe viral encephalitis.Twenty healthy age matched controls (10 cases of epilepsy and 10 cases of congenital abnormality) were also taken for the study.Cerebrospinal fluid exam,EEG,head MRI and other tests were performed in all patients.Caveolin-1,IL-1β and VEGF levels in cerebrospinal fluid of 65 children with viral encephalitis and 20 age-matched controls were measured using ELISA.Results Cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels in the acute phase of viral encephalitis were (49.209 ± 22.320) pg/ml,(16.923 ± 6.823) ng/ml,(44.342 ± 19.264) ng/ml respectively,and (33.253 ± 20.349)pg/ml,(11.724 ± 3.009)ng/ml,(30.312 ± 18.147) ng/ml in recovery phase,which were significantly higher than those of controls (P ＜0.01).The difference was statistically significant between acute phase and recovery phase (P ＜ 0.05).Acute viral encephalitis patients had higher Caveolin-l,IL-1β,VEGF levels than the epilepsy group,and the difference was statistically significant (P ＜ 0.05).In viral encephalitis group,children with cerebrospinal fluid protein content (0.5 ～ 1.0 g / L) had higher of Caveolin-1,IL-1β and VEGF levels as compared with those with cerebrospinal fluid protein content ≤ 0.5 g/L,and the difference was statistically significant (P ＜ 0.01).Cerebrospinal fluid Caveolin-1,IL-1 β and VEGF showed no significant difference among children with different severity of encephalitis,different levels of frequent seizures,different degrees EEG changes (P ＞ 0.05).But in the patients with severe head MRI changes,cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels increased significantly (P ＜ 0.05).Conclusions Caveolin-1,IL-1β and VEGF may participate in the pathogenesis of viral encephalitis.Detection of these parameters may be helpful to the evaluation of the severity and prognosis of viral encephalitis.