Objective:To explore the clinical characteristics and genetic variant in a premature infant with Netherton syndrome (NS).Methods:A neonate with NS caused by variants of SPINK5 gene diagnosed at the Children′s Hospital Affiliated to Zhejiang University School of Medicine in March 2020 was selected as the study subject. Clinical data and family history were collected. Peripheral blood samples (2 mL each) were obtained from the child and her parents for whole-exome sequencing (WES). Candidate variants were subjected to pathogenicity classification and deleteriousness evaluation. This study has been approved by the Medical Ethics Committee of the Hospital (Ethics No. 2024-IRB-0251-P-01). Results:The infant was born prematurely at 35 + 3 weeks due to "premature rupture of membranes for 4 hours" and exhibited generalized skin peeling, with meconium-stained amniotic fluid resembling bean curd residue. The condition improved with supportive treatments such as anti-infection and moisturizing therapy, though periodic hair loss had persisted. No similar case was reported by family history. WES has revealed a heterozygous c. 1130delG (p.G377Efs*127) variant in exon 14 of the SPINK5 gene, which was inherited from her mother, and deletion of exons 1 ~ 33 of the SPINK5 gene, which was inherited from her father. Conclusion:This case of NS presented with intrauterine onset in a preterm infant, which has not been previously reported. The identification of c. 1130delG (p.G377Efs*127) variant has expanded the mutation spectrum of the SPINK5 gene.

OBJECTIVE: To explore the clinical characteristics and genetic variant in a premature infant with Netherton syndrome (NS). METHODS: A neonate with NS caused by variants of SPINK5 gene diagnosed at the Children's Hospital Affiliated to Zhejiang University School of Medicine in March 2020 was selected as the study subject. Clinical data and family history were collected. Peripheral blood samples (2 mL each) were obtained from the child and her parents for whole-exome sequencing (WES). Candidate variants were subjected to pathogenicity classification and deleteriousness evaluation. This study has been approved by the Medical Ethics Committee of the Hospital (Ethics No. 2024-IRB-0251-P-01). RESULTS: The infant was born prematurely at 35⁺³ weeks due to "premature rupture of membranes for 4 hours" and exhibited generalized skin peeling, with meconium-stained amniotic fluid resembling bean curd residue. The condition improved with supportive treatments such as anti-infection and moisturizing therapy, though periodic hair loss had persisted. No similar case was reported by family history. WES has revealed a heterozygous c.1130delG (p.G377Efs*127) variant in exon 14 of the SPINK5 gene, which was inherited from her mother, and deletion of exons 1 ~ 33 of the SPINK5 gene, which was inherited from her father. CONCLUSION This case of NS presented with intrauterine onset in a preterm infant, which has not been previously reported. The identification of c.1130delG (p.G377Efs*127) variant has expanded the mutation spectrum of the SPINK5 gene.
Humans ; Serine Peptidase Inhibitor Kazal-Type 5/genetics* ; Netherton Syndrome/genetics* ; Female ; Infant, Newborn ; Infant, Premature ; Mutation ; Exome Sequencing ; Male

Objective:To explore the clinical characteristics and genetic variant in a premature infant with Netherton syndrome (NS).Methods:A neonate with NS caused by variants of SPINK5 gene diagnosed at the Children′s Hospital Affiliated to Zhejiang University School of Medicine in March 2020 was selected as the study subject. Clinical data and family history were collected. Peripheral blood samples (2 mL each) were obtained from the child and her parents for whole-exome sequencing (WES). Candidate variants were subjected to pathogenicity classification and deleteriousness evaluation. This study has been approved by the Medical Ethics Committee of the Hospital (Ethics No. 2024-IRB-0251-P-01). Results:The infant was born prematurely at 35 + 3 weeks due to "premature rupture of membranes for 4 hours" and exhibited generalized skin peeling, with meconium-stained amniotic fluid resembling bean curd residue. The condition improved with supportive treatments such as anti-infection and moisturizing therapy, though periodic hair loss had persisted. No similar case was reported by family history. WES has revealed a heterozygous c. 1130delG (p.G377Efs*127) variant in exon 14 of the SPINK5 gene, which was inherited from her mother, and deletion of exons 1 ~ 33 of the SPINK5 gene, which was inherited from her father. Conclusion:This case of NS presented with intrauterine onset in a preterm infant, which has not been previously reported. The identification of c. 1130delG (p.G377Efs*127) variant has expanded the mutation spectrum of the SPINK5 gene.

In order to provide basic information for the utilization and development of famous classical formulas containing Bletillae Rhizoma， this article systematically analyzes the historical evolution of the name， origin， harvesting and processing of Bletillae Rhizoma by reviewing the ancient materia medica， prescription books， medical books and modern literature. The research results showed that Baiji（白及） was the main name， some scholars took Baiji（白芨） as its main name， and there were many other names such as Baiji（白给）， Baigen（白根）， Baiji（白苙）. The mainstream source of Bletillae Rhizoma was the tubers of Bletilla striata， and drying， large， white， solid， root-free and skin removed completely were the good quality standards. With the promotion of wild to cultivated medicinal materials， there were certain differences between their traits， and the quality evaluation indexes should be adjusted accordingly. The origin of records in the past dynasties was widely distributed， with Guizhou and Sichuan having high production and good quality in modern times. The harvesting period is mostly in spring and autumn， and harvested in autumn was better. The processing and processing technology is relatively simple， and it was used fresh or powdered in past dynasties， while it is mainly sliced for raw use in modern times. Based on the results， it is suggested that the tubers of Bletilla striata of Orchidaceae should be used in the famous classical formulas， and it should be uniformly written as Baiji（白及）. And if the original formula indicates the requirement of processing， it should be operated according to the requirement， if the requirement of processing is not indicated， it can be used in raw form as medicine.

Objective To investigate the relationship between serum micro RNA(miRNAs)-873 and micro RNA-138-5p expression and tumor immune microenvironment(TIME)and prognosis in patients with non-small cell lung cancer(NSCLC).Methods A total of 108 NSCLC patients(NSCLC group)and 65 healthy volunteers(control group)who were admitted to Ba'nan Hospital Affiliated to Chongqing Medical University from February 2019 to February 2021 were selected.Real-time quantitative fluorescence polymeric chain reaction(qRT-PCR)was used to detect the expression of miR-873 and miR-138-5p in serum,and multiple immunofluorescence staining was used to detect tumor immune microenvironment indicators.Regular follow-up was conducted after discharge.Pearson analyzed the correlation between the expression of miR-873 and miR-138-5p in serum and the TIME index,and Kaplan-Meier and COX proportional risk regression analyzed the relationship between miR-873 and miR-138-5p and the prognosis of NSCLC patients.Results Comparison with control group,the expressions of miR-873(1.02±0.23 vs 3.15±0.82)and miR-138-5p(1.21±0.26 vs 3.54±0.92)in serum of NSCLC group were decreased,and the differences were statistically significant(t=-25.426,-24.769,all P＜0.05).The expressions of serum miR-873 and miR-138-5p of patients with low-to-moderate differentiation in TNM stages Ⅲ to Ⅳ were lower than those with highly differentiated patients in TNM stages Ⅰ to Ⅱ(t=9.615,10.253;6.889,3.361,all P＜0.05).The expressions of miR-873 and miR-138-5p in serum were negatively correlated with the values of PD-1,PD-L1,CD4 and CD8 H(r=-0.418～-0.673,all P＜0.05).The OS survival rate of NSCLC patients with low expression of miR-873 and miR-138-5p was lower than that of those with high expression of miR-873 and miR-138-5p(Log-Rankχ2=4.724,5.607,P＜0.05).TNM stage Ⅲ～Ⅳ was a risk factor for poor prognosis in patients with NSCLC(P＜0.05),and miR-873 and miR-138-5p were protective factors(P＜0.05).Conclusion The expressions of miR-873 and miR-138-5p in serum of NSCLC patients are down-regulated,which is related to TIME and low survival rate.

Objective To analyze the clinicopathological features of bladder urothelial carcinoma BUC in children and to share the experience of diagnosis and treatment.Methods The clinicopathological data,diagnosis and treatment process of a rare 9-year-old boy with BUC treated in the Second People's Hospital of Kashgar were reported and the literature was reviewed.Results The main clinical symptom is hematuria.Both preoperative urinary ultrasound and CT urography(CTU)identified the tumor,which was then removed with transurethral holmium laser resection under general anesthesia.Postoperative pathology confirmed papillary urothelial neoplasm of low malignant potential(PUNLMP).Epirubicin hydrochloride instillation was administered,and no evidence of recurrence or progress was detected during the 3-month follow-up.Conclusion Bladder urothelial carcinoma in children is extremely rare.The most common symptom is hematuria,and there are no characteristic signs.Urinary ultrasound is the first choice for preoperative imaging,and transurethral resection of the carcinoma results in good recovery.There is no recommendable instillation,and patients need long-term postoperative follow-up.

ObjectiveTo analyze the correlation between personality characteristics and family environment of undergraduate medical students taking psychology course. MethodsIn June 2017， a cross-sectional survey was conducted among 67 undergraduate medical students taking psychology course， and all the selected individuals were assessed using Minnesota Multiphasic Personality Inventory （MMPI） and Family Environment Scale-Chinese Version （FES-CV）. Then Pearson correlation analysis was performed between the original scores of MMPI subscales and the scores of FES-CV subscales. ResultsThe scores of hypochondriasis， depression， hysteria， psychopathic deviate， masculine-femininity， paranoia， psychasthenia and schizophrenia in MMPI of undergraduates were all higher than those of the national norm， with statistical differences （t=2.070~8.350， P<0.05 or 0.01）. The scores of cohesion， intellectual-cultural orientation， organization and achievement orientation in FES-CV of undergraduates were all lower than those of the national norm， while the scores of conflict tendency was higher than that of the national norm， with statistical differences （P<0.05 or 0.01）. Correlation analysis showed that cohesion score in FES-CV was negatively correlated with the scores of correction， hypochondriasis， depression and hysteria in MMPI （r=-0.410~-0.331， P<0.01）. Conflict tendency score was positively correlated with the scores of hysteria and psychasthenia （r=0.315， 0.415， P<0.01）. Active-recreational orientation score was negatively correlated with hypochondriasis score （r=-0.367， P<0.01） and positively correlated with schizophrenia score （r=0.346， P<0.01）. Control score was positively correlated with hysteria score （r=0.366， P<0.01）. ConclusionUndergraduate medical students taking psychology courses exhibit neuropathic and paranoid personality characteristics， and the personality characteristics are closely related to family environment.

Objective To observe the influences on prognoses and airway acid and base levels while using anti-inflammatory and anti-reflux therapies combined with "Hezhong Fuzheng massage" for treatment of infants with gastric volvulus (GV) and gastroesophageal reflux (GER)-induced pneumonia in order to provide evidence for clinical diagnosis and treatment of such infant disease. Methods Sixty infants 1-6 months old admitted to the Department of Integrated Traditional Chinese and Western Medicine of Wuhan Children's Hospital from January 2013 to December 2015 were diagnosed as pneumonia combined with GV and GER by the chest radiograph and radiography of upper gastrointestinal tract (UGT) with iodine, and according to difference in diagnostic methods, they were divided into an observation 1 group (30 cases) and an observation 2 group (30 cases). In observation 1 group, 24-hour pH value in upper digestive tract especially the distal esophagus was dynamically monitored, while in observation 2 group, 24-hour multichannel intraluminal impedance (MII) combined with pH monitoring of esophagus was carried out. Furthermore, two control groups were set up, each 30 cases; after radiography of the UGT with iodine, the control 1 group was diagnosed as GV and GER without pneumonia and control 2 group was diagnosed as only simple GV. Treatment of infant pneumonia was carried out in accordance with the guidelines for the management of community-acquired pneumonia in children; the treatment of GV and GER included postural, dietary, prokinetic and Hezhong Fuzheng massage [acupoint selection and massage was undertaken in 5 steps: push from the palmar crease to Banmen (rectilinear pushing manipulation) 300 times, push abdominal Yin and Yang (finger-pushing massage) 200 times, palpate the abdomen (clockwise) 100 times, poking of Tianshu 100 times, poke alternately bilateral Zusanli 100 times. Once-daily massage, (15±2) minutes each time, for consecutive 7 days]. The results of pH monitoring of the distal esophagus in observation 1 group was recorded; All GER data concerning pH monitoring of observation 2 group and the results of esophagus multi-channel intra-luminal impedance combined with pH monitoring in control group were compared. After 7 days of continuous treatment, the clinical efficacy of pneumonia and GV, hospitalization time, prognosis and the changes of sputum pH before and after treatment in the two observation groups were compared. Results In two observation groups, the main type of 60 cases with GV was organoaxial volvulus, accounting for 91.67% (55 cases), and more than 60% patients exhibited sputum pH <7.0. The distal esophagus 24-hour pH dynamic monitoring in observation 1 group showed that there were 26 cases with acid reflux (86.67%), 4 cases with GER negative (13.33%), the proportion of pH < 4 in GER negative patients was significantly lower than that in moderate acid reflux patients [3.35% (0.77% - 8.08%) vs. 26.23% (15.19% - 42.87%), P < 0.05], the number of long reflux (> 5 minutes) in GER negative patients was significantly reduced than that in mild acid reflux patients [times: 2 (0-5) vs. 7 (2-15), P < 0.05], the longest time of reflux in GER negative patients was significantly shorter than that in either mild or moderate acid reflux patients [minutes: 5.9 (2.5-10.0) vs. 19.2 (5.9-51.0), 41.6 (16.9-121.0), both P < 0.05]. The 24-hour MII-pH monitoring of esophagus in observation 2 group showed that there were 30 cases with pathological reflux, mainly mild acid reflux accounting for 90%, in which the percentage of proximal reflux events was 46.07% in the total reflux events. The numbers of acid reflux and proximal reflux in the observation 2 group were significantly higher than those in the control 1 and 2 groups [1 305 (37.72%) vs. 795 (25.69%) and 136 (18.89%), 1 594 (46.07%) vs. 687 (22.20%) and 154 (21.39%), both P < 0.05]; there were no significant differences in total effective rate (100% vs. 100%, P > 0.05) and hospitalization time (days: 7.58±1.09 vs. 7.67±1.12, P > 0.05) between the two observation groups. Conclusions For the diagnosis of infant pneumonia combined with GV and GER, the first selection of chest radiograph and radiography of the UGT with iodine can identify the severity of pneumonia, whether it is complicated with GV and its classification, and whether GER exists at the same time. The estimation of 24-hour pH dynamic monitoring of the acid reflux in the upper digestive tract especially the distal esophagus is consistent with that of the 24-hour MII-PH monitoring of esophagus. Moreover, 24-hour MII-PH monitoring of esophagus can also identify non-acid reflux, resulting in the GER diagnosis more accurate. Since the proximal reflux ratio of infants with GV combined with GER and pneumonia is relatively high and easy to cause cough and aspiration. attention should be paid on early diagnosis and timely intervention to such patients. The sputum acidity test can reflect the airway acid-base level and its manipulation is simple, so by that the real time disease situation can be estimated, and aspiration of sputum also can help the treatment. The therapeutic schedule formulated by our group can elevate the therapeutic effect, improve the airway acid-base environment and benefit prognosis.

Objective: To establish real-time recombinase polymerase amplification（RPA）for the rapid detection of Vibrio parahaemolyticus（VP）. Methods: An exo probe and primers were designed according to the conserved sequence of thermolabile hemolysin（tlh）gene of VP and then RPA for detection of VP was established. The sensitivity of the assay was evaluated by detecting different concentration of VP;the specificity was evaluated by detecting different bacteria;the stability was evaluated by repeat trials;the application effect was evaluated by detecting food samples which were simultaneously tested with traditional culture method according to GB 4798.7-2013 Detection of VP. Results: A real-time RPA was established to complete VP amplification within 20 min at a constant temperature of 39 ℃. The analytical sensitivity of the assay was five pg per reaction and no cross-reactivity with other pathogenic bacteria observed. The RPA detection results with different concentration of VP and E. coli DNA templates at three time points were consistent. The detection results of 51 food samples by RPA were the same as those by traditional culture method. Conclusion The established real-time RPA can qualitatively detect VP,with simple operation and interpretation of results,which is suitable for rapid detection of VP in public health emergencies and food safety supervision.

Objective To investigate feasible immunotherapy strategy using tumor specific cell against osteosarcoma,and to analyze the therapeutic effect of adoptive cellular infusion therapy on osteosarcoma.Methods Decitabine (DAC) was employed as a hypomethylating agent for the treatment in osteosarcoma cell lines HOS and U2OS.After treatment,the expression of cancer-testis antigen (CTA) was evaluated by PCR and Western Blot.In animal studies,human osteosarcoma cell line HOS,which was transfected by luciferase and HLA-A0201 in previous,was inoculated into immune deficient NOD-SCID mice to establish osteosarcoma xenografts.Ex-vivo expanded CTA specific homo CD8+T-ells were labeled with DiR and injected into the mice via the tail vein.In vivo imaging system was utilized to detect the distribution of administrated CD8+ T-cells.In addition,the progression of tumor xenografts was monitored.Moreover,mouse K7M2 osteosarcoma cell line was used to establish animal models in immune competent BALB/c mice.Immune competent models were utilized to evaluate the effectiveness of hypomethylating treatment in regarding to spontaneous immune attack against tumors.Flow cytometry was used to analyze the proportion of intratumoral lymphocytes and the status of these effector antitumor immune cells,and to reveal the effect of hypomethylating treatment in facilitating lymphocyte infiltration and activation.Results The expression of all the evaluated cancer/testis antigens were elevated in HOS and U2OS osteosarcoma cell lines after hypomethylating treatment with DAC.The proliferation of in vitro cultured osteosarcoma cells can be significantly suppressed after at least 5 d treatment with DAC.Besides,DAC alone controlled osteosarcoma cell proliferation.In immune deficient mouse models,hypomethylating pre-treatment resulted in successful T-cell homing to tumor sites.Moreover,the combination treatment with DAC and CTA specific T-cell adoptive transfer significantly suppressed tumor proliferation.In immune competent mouse models,hypomethylating treatment with DAC improved autologous T-cell infiltration into the tumor,and strengthened the activity of intratumoral CD8+ T-cells,elevated the secretion of IFN-gamma,granzyme B and perforin by CD8+ T-cells.Conclusion Hypomethylating treatment is able to suppress osteosarcoma cell proliferation,improve the expression of CTA in osteosarcoma cells,and consequently provide optimal environment for CTA specific T-cell adoptive therapy.