Objective To detect the presence of the apoptotic sensory neurons in rat dorsal root ganglion (DRG) following non-freezing cold injury of the sciatic nerve.Methods Thirty three male Wistar rats were randomly divided into three groups (7 d group, 14 d group, 21 d group; n=11 each).The sciatic nerves were cooled to 4 ℃ for 2 hours and the control side underwent sham operation.The pathological examination was performed on L4-6 DRGs at 7, 14, and 21 d post operation.The apoptotic sensory neurons of L4-6 DRGs were detected using FCM (Annexin /PI) and TUNEL staining.Results The apoptotic sensory neurons of L4-6 DRGs were revealed after non-freezing cold injury of the rat sciatic nerve.FCM (Annexin /PI) and TUNEL quantitative analysis indicated that the apoptotic neurons significantly increased in rat dorsal root ganglion (DRG) at 7 d, 14 d, and 21 d post operation following non-freezing cold injury of the sciatic nerve.TUNEL qualitative analysis further confirmed pathological characters of neuron apoptosis in L4-6 DRGs.The apoptotic neurons began to increase from 7~(th) day, reached peak at 14~(th) day, and then decreased slightly at 21 d following operation.Conclusions Non-freezing cold injury on sciatic nerves can cause the sensory neurons apoptosis of L4-6 DRGs.

Objective To investigate the morphological changes in the sciatic nerve and the dorsal root ganglions (DRGs) and also gene expression in DRGs after non-freezing cold injury, and to explore the molecular mechanism of peripheral nerve cold injury and regeneration. Methods Twenty-four male Wistar rats were used. The sciatic nerve on one side was cooled to 4℃ for 2 h, and the sciatic nerve on the opposite side was exposed, but without cooling. Sciatic nerves and L4, L5 and L6 DRGs from both sides were harvested at the 1st, 2nd and 3rd week after cooling. Any pathological changes were observed using light and electron microscopy. Laser capture microdissection (LCM) was used to investigate the DRG neurons' gene expression. The array result was verified with RT-PCR for eight genes. Results Large fiber degeneration was obvious by the 7th day after cooling. Myelinated fiber regeneration had begun by the 14th day, so this time was chosen to explore the neurons' gene expression. Ninety-six genes and expressed sequence tags (ESTs) were up-regulated greater than 2 fold. Their proteins' functions were classified as adaptive response to external stimulus, apoptosis regulation, cell adhesion, immune and inflammation response,nerve regeneration, pain associated molecules, microtubule cytoskeleton, ion-channels, neurotransmitters and receptors, and neuropeptides. Conclusions A complex molecular mechanism is involved in cold injury and regeneration of the sciatic nerve, and many genes are involved. Large scale microarray analysis is a potent means to screen out related genes, thus suggesting future repair strategies.

Objective Investigate the prognosis of patients undergoing liver transplantation (LT) for end-stage autoirnmune liver disease (ALD).Method The clinical data of 45 patients with endstage ALD undergoing LT from April 2001 to March 2015 in the first affiliated hospital of Zhejiang University were analyzed retrospectively.The postoperative cumulative survival rate of the recipients was calculated,and the causes of death were analyzed.The postoperative rejections,new onset viral hepatitis and ALD recurrence were also analyzed.Result In 45 ALD recipients,33 cases survived and the postoperative 5-year cumulative survival rate was 78.8％.Causes for 12 dead cases were mnultiple organ failure,liver graft failure,respiratory complications,hemorrhage and hepatic artery embolization.In 45 ALD recipients,6 cases suffered rejection after operation with the incidence bing 13.3％.One case suffered new onset hepatitis B infection 8 years after opcration.One recipient suffered primary disease (primary biliary cirrhosis) recurrence 2 years after operation,and 1 recipient with primary disease (primary sclerosing cholangitis) developed into overlap syndrome.They all survived for a long term after active treatments.Conclusion Most LT recipients with endstage ALD can obtain a long term survival.Attentions should be paid to the immunosuppressive regimens in early period after LT,prevention of infection,rejection and postoperative new onset viral hepatitis,and timely diagnosis of primary disease recurrence.

Objective To assess the therapeutic effects of flupentixol and melitracen tablets combined with Saccharomyces boulardii on patients with diarrhea-predominant irritable bowel syndrome (IBS) accompanied with anxiety and depression.Methods This multi-center,randomized,prospective study enrolled 84 patients with diarrhea-predominant IBS who were divided into combined treatment group (42 patients) and control group (42 patients).Saccharomyces boulardii was administrated in both of the groups,and flupentixol and melitracen was added in combined treatment group.The treatment course was four weeks.The gastrointestinal symptoms and mood disorders were evaluated before treatment,one week and four weeks after treatment.Adverse reactions were also observed.Chi-square test was performed for statistical analysis.Results At the end of one week after treatment,the efficacy rates of gastrointestinal symptoms improvement of combined treatment group and control group were 31.0％ (13/42) and 23.8％ (10/42),and there was no statistically significant difference (P＞0.05).At the end of four weeks after treatment,the efficacy rate of gastrointestinal symptoms improvement of combined treatment group was 92.5％ (37/40),which was higher than that of control group (73.2％,30/41),and the difference was statistically significant (x2 =5.291,P =0.037).At the end of one week after treatment,the efficacy rates of Hamilton Depression Scale score improvement of combined treatment group and control group were 31.6％ (12/38) and 12.1％ (4/33),and there was no statistically significant difference (P＞0.05).At the end of four weeks after treatment,the efficacy rates of Hamilton Depression Scale score improvement of combined treatment group was 63.9％ (23/36),which was higher than that of control group (34.4％,11/42),and the difference was statistically significant (x2 =6.433,P=0.043).At the end of one week and four weeks after treatment,the efficacy rates of Hamilton Depression Scale score improvement of combined treatment group were 35.7％ (15/42) and 80.0％ (32/40),which were higher than those of control group (15.4％,6/39 and 34.2％,13/38),and the differences were statistically significant (x2 =9.759,P=0.007; x2 =17.105,P＜0.01).One week after treatment,the adverse events rates of combined treatment group and control group were 4.8％ (2/42) and 4.8％ (2/42) ; four weeks after treatment,the adverse events rates of combined treatment group and control group were 2.5％ (1/40) and 2.4％ (1/41).There was no statistically significant difference in adverse events rates between two groups (both P＞0.05).Conclusions Flupentixol and melitracen combined with Saccharomyces boulardii treatment could not only improve the anxiety and depression symptoms of patients with diarrhea-predominant IBS,but also effectively improve gastrointestinal symptoms.The efficacy of combined treatment is better than monotherapy Saccharomyces boulardii alone treatment.

OBJECTIVETo examine the feasibility and safety of gastric submucosal tunnel dissection of gastric submucosal tumors (SMTs) by double tunnel and double flex endoscope.

METHODSFifty patients with gastric SMTs detected by gastric endoscopy and endoscopic ultrasonography between January, 2012 and August, 2013 were enrolled in this study. Using carbon dioxide throughout the procedure, the mucous in the arc was incised along the margins of the lesion to separate the submucosa and create a tunnel. The exposed SMTs were resected completely and the mucosa was covered by endoscopic forceps followed by clipping of the incision. The complication, clinical outcomes, hospital stays and operation time were evaluated.

RESULTSOf the 50 lesions, 50 were located in the gastric fundus, 17 in the gastric antrum and 5 in the gastric body. The lesions were completely resected in all the patients. The diameter of the resected lesions ranged from 0.5 to 2.5 cm (mean 1.1 ± 0.6 cm), and the operation lasted for 35.3 ± 16.2 min (range 23-76 min). In 5 cases (10%), perforation occurred during the operation and was closed by clipping the incision with endoclips after the lesion resection; these patients were discharged after conservative management. Intraoperative bleeding occurred in 16 cases and was successfully managed through endoscopic methods. No delayed postoperative bleeding or perforation occurred in these patients. None of the 48 patients followed up showed tumor recurrence at one year after the operation, and 2 patients were lost for follow up.

CONCLUSIONEndoscopic submucosal dissection of gastric SMTs is effective and safe using double tunnel and double flex endoscope.

Dissection ; Endoscopes ; Endoscopy ; Endosonography ; Gastric Mucosa ; pathology ; surgery ; Humans ; Neoplasm Recurrence, Local ; Stomach Neoplasms ; surgery

Objective To detect retinoid X receptor α (RXRα) mRNA expression in blood of subjects exposed to different concentrations of arsenic via drinking water, to analyze the relationship between RXRα mRNA expression and skin lesion caused by arsenic,and further to explore the skin lesion mechanism of arsenic. Methods Study sites were selected by molecular epidemiology method from high arsenic drinking water area of Bayannur City. Two hundred and thirty-five subjects who had been lived in high arsenic area for more than 10 years were selected;blood samples and water samples were collected from the subjects; according to arsenic concentration in drinking water,they were divided into four groups,<10 μg/L(control group),10-<100 μg/L(low dose group),100- <200 μg/L (middle dose group), and ≥200 μg/L (high dose group). Skin hyperkeratosis and pigment abnormity examination were conducted. The RXRα mRNA expression level in blood samples was detected by real-time quantitative PCR, and then the relationship between expression of RXRα mRNA and different levels of arsenic exposure,and skin lesion induced by arsenic were analyzed. Results ①The results showed that there was a dose-effect relationship between the prevalence of hyperkeratosis, pigment abnormity and arsenic exposure (χ2= 14.597, 12.825, P < 0.05); ②With increasing of arsenic exposure, RXRα mRNA expression in blood decreased firstly and then increased (F = 8.312, P < 0.05), which were significantly different statistically from those of control [(1.20 ±0.53)×10-3]and low dose groups[(0.92 ± 0.49)×10-3,P<0.05];RXRα expression was significantly higher in high dose group[(1.40 ± 0.45)×10-3]than those of middle and low dose groups [(1.12 ± 0.58,0.92 ± 0.49)×10-3,P<0.05]; ③The RXRα mRNA expression in people with different level of skin damage (hyper keratosis and pigment abnormity)were statistically significant(F=4.206,4.389, P< 0.05); degree Ⅲ[(1.98 ± 0.38) × 10-3] hyperkeratosis patients compared with degree Ⅰ [(1.11 ± 0.52) × 10-3] and degree Ⅱ [(1.13 ± 0.42) × 10-3], RXRα mRNA expression was significantly different (P < 0.05), degree Ⅱ and higher degrees [(1.61 ± 0.54) × 10 -3] pigment abnormity patients compared with control [(1.15 ± 0.52)×10-3],RXRα mRNA expression was significantly different (P < 0.05). Conclusions Chronic arsenic exposure has an effect on RXRα mRNA expression in blood. There is a relationship between abnormal expression of RXRα mRNA and skin lesion induced by arsenic.

Objective To understand the changes of skin lesions in population exposed to arsenic through drinking-water in Inner Mongolia after changing water source for 12 years,and to evaluate the long-term harmful effects and the delayed injury to human body due to arsenic exposure and the effect of changing water.Methods The stratified random cluster sampling investigation objects (data derived from the Inner Mongolia Autonomous Region Comprehensive Center for Disease Control and Prevention) of the arsenic exposure area before water reform (2004) as the foundation,in 2017,we selected three administrative villages (changed water in 2005) in Bayan Nur City of Inner Mongolia as survey sites.The objects of this study were residents who had been living in the survey site and were eligible for previous survey (n =80,35 males and 45 females).To compare the skin damage of the residents exposed to arsenic before and after the water changed,the water samples of the survey objects were measured arsenic content.According to the content of water arsenic,the respondents were divided into low,medium and high exposure groups (10-,150-,≥300 μg/L),to analyze clinical classification in skin damage before and after changing water.Results The water arsenic content after changing [1.42 (0.18-33.45) μg/L] was lower than those before the changes [238.20 (14.56-824.70) μg/L,Z =-8.34,P ＜ 0.05].A total of 63 persons with skin keratinization,7 persons with hyperpigmentation and 19 persons with depigmentation were identified after the changes of drinking water source,while 38,3 and 18 persons were respectively found before the changes.The detection rate of skin keratinization after water changes was significantly higher than that before water changes [78.8％ (63/80) vs 47.5％ (38/80),x2 =16.78,P ＜0.05].According to the clinical classification of skin damage,23 patients were normal,44 patients were suspicious,10 patients were mild,and 3 patients were moderate or severe after the water changes,compared with those before the water changes (38,18,6 and 18 persons were respectively found),the clinical fractional suspicious detection rate of skin damage in the arsenic exposed population increased,and the medium-severe detection rate decreased,and the differences were statistically significant (x2 =17.80,12.33,P ＜ 0.05).The detection rate of the clinical score of moderate-severe skin damage in men was significantly decreased,and the difference was statistically significant (x2 =7.65,P ＜ 0.05).The normal detection rate of female skin lesions was reduced,and the rate of suspected detection was increased (x2 =5.48,7.65,P ＜ 0.05).In the high-and medium-dose groups,41.9％ (13/31) and 42.9％(12/28) of the arsenic exposure population had a reduced clinical classification of skin damage.The ratios of clinical classification severity of skin damage in the high-,medium-and low-dose groups were 25.8％ (8/31),25.0％ (7/28),and 42.9％ (9/21).The differences were statistical significantly by linear trend chi-square test (x2 =12.96,P ＜ 0.05).Conclusions The skin lesions exposed to arsenic could be effectively improved after changing water.But the skin lesions still appear in some cases due to long-term chronic arsenic exposure.The long-term effects caused by arsenic should be explored persistently.

Objective: Excessive daytime sleepiness (EDS) and insomnia symptoms are common in patients with major depressive disorder (MDD), which might lead to a poor prognosis and an increased risk of depression relapse. The current study aimed to investigate the prevalence, and sociodemographic and clinical correlates of EDS and insomnia symptoms among adolescents with MDD. Methods: The sample of this cross-sectional study included 297 adolescents (mean age=15.26 years; range=12–18 years; 218 females) with MDD recruited from three general and four psychiatric hospitals in five cities (Hefei, Bengbu, Fuyang, Suzhou, and Ma’anshan) in Anhui Province, China between January and August, 2021. EDS and insomnia symptoms, and clinical severity of depressive symptoms were assessed using Epworth sleepiness scale, Insomnia Severity Index, and Clinical Global Impression-Severity. Results: The prevalence of EDS and insomnia symptoms in adolescents with MDD was 39.7% and 38.0%, respectively. Binary logistic regression analyses showed that EDS symptoms were significantly associated with higher body mass index (odds ratio [OR]=1.097, 95% confidence interval [CI]=1.027–1.172), more severe depressive symptoms (OR=1.313, 95% CI=1.028–1.679), and selective serotonin reuptake inhibitors use (OR=2.078, 95% CI=1.199–3.601). And insomnia symptoms were positively associated with female sex (OR=1.955, 95% CI=1.052–3.633), suicide attempts (OR=1.765, 95% CI=1.037–3.005), more severe depressive symptoms (OR=2.031, 95% CI=1.523–2.709), and negatively associated with antipsychotics use (OR=0.433, 95% CI=0.196–0.952). Conclusion EDS and insomnia symptoms are common among adolescents with MDD. Considering their negative effects on the clinical prognosis, regular screening and clinical managements should be developed for this patient population.

Objective To analyze the clinicopathological features and lymph node metastases characteristics of intrahepatic cholangiocarcinoma (ICC).Methods The retrospective case-control study was conducted.The clinicopathological data of 1 321 ICC patients who were admitted to 12 hospitals from April 2007 to November 2017 were collected,including 182 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,173 in the First Affiliated Hospital of Xi'an Jiaotong University,156 in the First Hospital Affiliated to Army Medical University (Third Military Medical University),139 in the Cancer Center of Sun Yatsen University,128 in the Zhongshan Hospital of Fudan University,113 in the Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine,109 in the First Affiliated Hospital of Dalian Medical University,84 in the Tianjin Medical University Cancer Institute and Hospital,65 in the First Affiliated Hospital of Zhengzhou University,62 in the Affiliated Hospital of North Sichuan Medical College,58 in the Affiliated Drum Tower Hospital of Nanjing University Medical School and 52 in the Second Affiliated Hospital of Zhejiang University School of Medicine.Treatment planning was respectively determined by comprehensive hospitals according to clinical features and results of related examinations.Surgical procedures were performed based on the results of intraoperative examinations and patients' conditions.Observation indicators:(1) laboratory examination and treatment situations;(2) relationship between clinicopathological features and primary tumor location and diameter;(3) diagnosis of lymph node dissection and metastases;(4) relationship between clinicopathological features and lymph node metastases.Measurement data with normal distribution were represented as x±s,and comparison between groups was done by the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the chi-square test.Results (1) Laboratory examination and treatment situations:of 1 321 patients,cases and percentages of positive hepatitis B virus (HBV),positive serum alpha-fetoprotein (AFP) (＞ 20 μg/L),positive serum carcinoembryonic antigen (CEA) (＞ 5 μg/L),positive serum alanine transaminase (ALT) (＞75 U/L),positive serum CA19-9 (＞37 U/mL) and positive serum total bilirubin (TBil) (＞20 μmol/L) were respectively 202,80,329,207,590,300 and 15.586％ (202/1 296),7.339％(80/1 090),26.299％ (329/1 251),16.587％ (207/1 248),49.789％(590/1 185),24.000％ (300/1 250).Of 1 321 patients,1 206 underwent surgery,including 904 with hepatectomy,193 with transcatheter arterial chemoembolization (TACE),72 with percutaneous transhepatic biliary drainage (PTBD),3 with endoscopic retrograde cholangiopancreatography (ERCP),3 with PTBD + ERCP and 31 with other treatments;115 had missing registration of surgical procedures.(2) Relationship between clinicopathological features and primary tumor location and diameter:① Relationship between clinicopathological features and primary tumor location:cases with CA19-9 level ≤ 37 U/mL,38-200 U/mL and ＞ 200 U/mL were respectively 227,91,146 with primary tumor located in left liver and 282,134,137 with primary tumor located in right liver.Cases combined with hepatitis and intrahepatic bile duct stone were respectively 67,73 with primary tumor in left liver and 111,47 with primary tumor in right liver,with statistically significant differences in above indictors (x2 =6.710,5.656,12.534,P＜0.05).The results of further analysis showed that incidence age of ICC was (63± 10)years old in patients with hepatitis and (59± 10) years old in patients without hepatitis,with statistically significant differences (t =4.840,P＜0.05).② Relationship between clinicopathological features and primary tumor diameter:cases with primary tumor diameter ≤ 3 cm,with 3 cm ＜ primary tumor diameter ≤ 5 cm and with primary tumor diameter ＞ 5 cm were respectively 159,250,229 with CEA level ≤ 5 pg/L and 40,65,95 with CEA level ＞ 5 μg/L and 165,258,286 with ALT level ≤ 75 U/L and 34,57,36 with ALT level ＞ 75 U/L and 148,242,281 with TBil level ≤ 20 μmol/L and 51,73,43 with TBil level ＞ 20 μmol/L,and 37,70 and 131 patients had satellite loci of tumor,with statistically significant differences in above indictors (x2=8.669,6.637,15.129,34.746,P＜0.05).(3) Diagnosis of lymph node dissection and metastases:of 904 patients with hepatectomy,346 received lymph node dissection,total number,number in each patient and median number of lymph node dissected were respectively 1 894.0,5.5 and 4.0 (range,1.0-26.0);157 had lymph node metastases,with a rate of lymph node metastasis of 45.376％ (157/346),number and number in each patient of positive lymph node were respectively 393.0 and 2.5.Of 346 patients with lymph node dissection,114 had lymph node metastasis by preoperative imaging examination,postoperative pathological examinations confirmed 64 with lymph node metastasis and 50 without lymph node metastasis;232 didn't have lymph node metastasis by preoperative imaging examination,postoperative pathological examinations confirmed 93 with lymph node metastasis and 139 without lymph node metastasis.The sensitivity,specificity and diagnostic accuracy by imaging examination were respectively 40.8％,73.5％ and 58.7％.(4) Relationship between clinicopathological characteristics and lymph node metastases:lymph node metastasis rate was respectively 51.397％ (92/179) in patients with primary tumor located in left liver,39.103％ (61/156) in patients with primary tumor located in right liver,34.615％(18/52) in patients with primary tumor diameter ≤ 3 cm,42.500％(51/120) in patients with 3 cm ＜ primary tumor diameter ≤ 5 cm,52.980％(80/151) in patients with primary tumor diameter ＞ 5 cm,48.790％(121/248) in patients with CEA ≤ 5 μg/L,59.302％ (51/86) in patients with CEA ＞ 5 μg/L,40.234％ (103/256) in patients with TBil ≤ 20 μmol/L,35.106％ (33/94) in patients with TBil ＞ 20 μmol/L,with statistically significant differences in above indictors (x2 =5.078,6.262,9.456,5.156,P ＜ 0.05).Conclusions The related etiological factors may be different due to different primary location of ICC.Primary tumor in left liver is significantly associated with intrahepatic bile duct stone of left liver,and primary tumor in right liver is associated with combined hepatitis,while combined hepatitis could lead early occurrence of ICC.At present,there is a low rate of lymph node dissection in ICC patients and a high lymph node metastasis rate in patients receiving lymph node dissection.Lymph node metastasis is associated with primary tumor location and diameter,levels of CEA and TBil.Lymph node metastasis rate in left liver is higher than that in right liver,and is getting higher with increased primary tumor diameter.

[Abstract] Objective: To study the expression of miRNA-95 in osteosarcoma tissues and cell lines, as well as to reveal its effect on proliferation, apoptosis, cell cycle and invasion ability of osteosarcoma cells. Methods: Real-time fluorescent quantitative PCR was used to detect the expression of miRNA-95 in 15 pairs of osteosarcoma tissues and their adjacent normal tissues (specimens were collected from patients underwent surgery in Qingdao Haici Medical Group from January 2015 to January 2018), osteosarcoma cell lines (MG-63, U2OS, 143B and HOS) and normal human osteoblast hFOB1.19 cell line. miRNA-95 mimics and miRNA-95 inhibitors were respectively transfected into MG-63 cells by Lipofectamine 2000, and miRNA-NC group was set up as control group. CCK-8 method was used to detect the changes in cell proliferation, Flow cytometry was used to detect the changes in cell cycle and apoptosis, Transwell method was used to detect the changes in cell invasion ability, and Dual luciferase enzyme activity assay was used to detect and validate the target gene of miRNA-95 in osteosarcoma cells. Results: The expression level of miRNA-95 in human osteosarcoma tissues and cell lines (MG-63, U2OS, 143B and HOS) was significantly higher than that in adjacent tissues and normal human osteoblast hFOB1.19 cell line (all P<0.01), with the highest expression in MG-63 cells (P<0.01). Compared with the miRNA-NC group, the proliferation and invasion abilities of MG-63 cells in miRNA-95 mimics group increased significantly, while the apoptosis rate decreased significantly (all P<0.01). However, the proliferation and invasion activities of MG-63 cells in miRNA-95 inhibitor group decreased significantly, while the apoptosis rate increased significantly, and the cell cycle was obviously blocked (all P<0.01). miRNA-95 played a role in targeting the gene of epithelialmembraneprotein1 (EMP-1) in human osteosarcoma MG-63 cells. Conclusion: miRNA-95 is highly expressed in human osteosarcoma tissues and cells; inhibitor of miRNA-95 expression can promote apoptosis and inhibit proliferation, cell cycle and invasion of osteosarcoma cells, which may be related with targeting EMP-1 gene.