[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .

This study was aimed to explore five Chinese medicine components or ingredients from Qu-Shi Hua-Y u Decoction (QSHYD) with uniform design method and screen ingredients assembling of Chinese medicine BZL pre-scription in order to verify its therapeutic effect on experimental fatty liver rats. High-fat diet was used in the estab-lishment of fatty liver rat models. Five effective ingredients (i.e., chlorogenic acid, geniposide, curcumin, polydatin and polysaccharide of Atractylodes macrocephala Koidz) of QSHYD were used as study subjects. Uniform design was applied in the grouping design. The intragastric administration was given for four weeks. The screening index was the content of liver triglyceride (TG). The ingredients assembling of Chinese medicine BZL prescription was screened through uniform design and regression analysis. The same fatty liver animal model was used in the comparison be-tween QSHYD and rosiglitazone. The TG content in liver tissues of rats and serum ALT activity were detected. The pathological changes of liver tissues were observed (HE stain and oil red O stain) with Ridit analysis to verify its therapeutic effect. The results showed that through screening, the regression equation was Y = 15.083X1 + 5.321X2- 5.186X3 - 16.157X4 + 9.35X5 + 17.667X3X4 - 8.422X1X2 - 6.617X3X5 + 16.571 (X1: chlorogenic acid, X2:polydatin, X3: polysaccharide of Atractylodes macrocephala Koidz, X4: geniposide, X5: curcumin). According to this equation, the best TG reducing effect occurred when the chlorogenic acid (X1), polysaccharide of Atractylodes macrocephala Koidz (X3), and geniposide (X4) were the maximum dosage. It showed that the best combination in inhibiting TG content in liver tissues was X3, X4 and X1 (i.e. BLZ prescription). Among fatty liver rat mod-els induced with same high-fat diet, the screened Chinese medicine ingredient BZL prescription can obviously reduce its TG content in liver tissues and serum ALT activity ( P < 0 . 01 ) . It obviously improved the fatty degen-eration of rat's liver. It was concluded that ingredients assembling of Chinese medicine BZL prescription screened through uniform design can obviously prevent and treat fat deposition and injury of the liver among rat models induced by high-fat diet .

Objective To identify the differences of circulating histone H4 (hH4) levels in different phrase of children under 1 year old with sepsis, investigating its possible role in the process of sepsis. Methods We retrospectively analyzed 40 sepsis patients aged between 1 and 12 months old who were ad-mitted in PICU of Shenzhen Maternity and Children Healthcare Hospital between November 2016 and December 2017. Their clinical data were reviewed,and then they were categorized into three groups,non-criti-cal group(n=21),critical group(n=11) and extremely critical group(n=8) by Pediatric Critical Illness Score ( PCIS) . Forty children of the same age group with no signs of infection were enrolled as control group. Plasma from 4 groups on admission day and the third day after admission were collected. Plasma hH4 levels of all samples were measured and all parameters were reconfigured and statistically analyzed. Results HH4 levels of samples were collected from sepsis group on admission day:non-critical group (97. 22 ± 16. 92)μg/L,criti-cal group (148. 09 ± 33. 00)μg/L,extremely critical group (195. 04 ± 45. 85)μg/L. HH4 levels of all three sub groups were significantly higher than those of control group [(46. 07 ± 18. 06)μg/L, H = 64. 16, P<0. 001]. Data of the third day were as follow:non-critical group (98. 96 ± 16. 29)μg/L,critical group (152. 57 ± 29. 04)μg/L,extremely critical group (239. 52 ± 49. 84)μg/L. Comparing the parameters of the first and the third admission day,hH4 levels of both non-critical group (Z= -1. 42,P=0. 16) and critical group (Z= -1. 48,P=0. 14) showed no significant difference. However,hH4 levels of extremely critical group were significantly increased on the third admission day (Z= -2. 67,P=0. 008). Setting the discharge day or the seventh day after admission as the endpoint of the observation, hH4 levels of the death group [241. 53(229. 19,245. 07)μg/L]were significantly higher than that of the survival group[115. 21(96. 38,136. 15)μg/L,Z= -2. 80,P=0. 005]. Among the sepsis,hH4 levels were negatively correlated with their PCIS results on admission day(r= -0. 853,P<0. 001). The hH4 levels were also negatively correlated with the patients'PCIS score changes on the third day after admission(r = -0. 554,P <0. 001). Conclusion The levels of circulating hH4 in sepsis children are co-related to the prognosis and severity suggesting that it may affect the process of the sepsis in certain ways.

This study details a rare instance of a high fibula fracture, classified as a Bosworth fracture. A 54-year-old male presented with pain and restricted movement in his right ankle after a sprain that occurred 10 hours ago. After initial assessments he was diagnosed as a right ankle fracture with posterior dislocation, a proximal right fibula fracture, and an injury to the right lower tibiofibular joint. According to Lauge-Hansen classification this was a type IV injury charactered by pronation-external rotation. Attempts at manual reduction were unsuccessful. Further evaluation using CT scans with three-dimensional reconstruction showed a separation and dislocation at the lower tibiofibular joint, with the distal fibula interlocked with the posterolateral tibia. Additionally, comminuted fractures of the medial and posterior malleoli showed significant displacement and misalignment, leading to persistent ankle dislocation. Immediate calcaneal traction and symptomatic treatment were applied. On the third day of admission a reevaluation indicated successful reduction of the dislocated ankle and tibiofibular joint. Surgery was then performed with favorable outcome. Follow-up X-rays at one and a half years after surgery demonstrated bony heal of the fractures with normal ankle joint space and functionality. Bosworth fractures involving pronation-external rotation are notably less common than those with posterior-external rotation. When manual reduction proves ineffective, applying continuous calcaneal traction with gradual tension can help unlock fibular dislocation, thereby expediting surgical intervention.

Objective:To discuss the safety and feasibility for the use of 3D uniportal VATS sleeve resection.Methods:Totally 32 patients with central lung cancer received 3D uniportal VATS sleeve resection(group A) from June 2017 to May 2020 at Shanghai Chest Hospital. Meanwhile, 63 patients received conventional VATS sleeve resection(group B). The clinicopathological and perioperative outcome data were retrospectively collected and analyzed.Results:The baseline clinicopathological characteristics between these two groups were statistically similar. Compared with group B, the mean operative time[(174.19±73.69)min vs.(212.46±50.02)min, P=0.004] and blood loss[(73.13±42.70)ml vs.(130.48±133.72)ml, P=0.020] of group A were decreased, harvested lymph node stations was increased(7.63±1.59 vs. 6.76±1.70, P=0.018). Lymph nodes dissected showed no statistical difference(1.31±1.58 vs 1.21±1.96, P=0.803). There was no intraoperative death in both groups. Inspiringly, group A possessed lower rate of conversion to thoracotomy(0 vs. 36.5%, P=0.000), shorter chest drainage durations[(4.88±1.15)days vs.(6.81±3.8)days, P=0.007]. Although there were no deaths during hospitalization in both groups, the incidence of postoperative complications in group A was significantly lower than that in group B(25.0% vs. 47.6%, P=0.046). It also presented more complicated operations including pulmonary artery plasty(25.0% vs. 6.3%, P=0.024) and carina plasty(12.5% vs. 1.6%, P=0.005) against group B. Conclusion:3D uniportal VATS was a safe and feasible technique for the surgical treatment of central lung cancer when conducting a thoracoscopic sleeve resection.

Background: Nasal?type extranodal natural killer/T?cell lymphoma (ENKTCL) originates primarily in the nasal cavity or extra?nasal sites within the upper aerodigestive tract. However, it is unclear whether the primary site can serve as an independent prognostic factor or whether the varying clinical outcomes observed with different primary sites can be attributed merely to their propensities of regional lymph node involvement. The aim of this study was to investigate the prognostic implications of the primary site and regional lymph node involvement in patients with early?stage nasal?type ENKTCL. Methods: To develop a nomogram, we reviewed the clinical data of 215 consecutively diagnosed patients with early?stage nasal?type ENKTCL who were treated in Sun Yat?sen University Cancer Center with chemotherapy and radiotherapy between 2000 and 2011. The predictive accuracy and discriminative ability of the nomogram were determined using a concordance index (C?index) and calibration curve. Results: The 5?year overall survival (OS) and progression?free survival (PFS) rates of patients with nasal ENKTCL were higher than those of patients with extra?nasal ENKTCL (OS: 68.2% vs. 46.0%, P = 0.030; PFS: 53.4% vs. 26.6%, P = 0.010).The 5?year OS and PFS rates of patients with Ann Arbor stage IE ENKTCL were higher than those of patients with Ann Arbor stage IIE ENKTCL (OS: 66.3% vs. 59.2%, P = 0.003; PFS: 51.4% vs. 40.3%, P = 0.009). Multivariate analysisshowed that age >60 years, ECOG performance status score nasal primary site, and regional lymph node involvement were significantly associated with lower 5?year OS rate;≥2, elevated lactate dehydrogenase (LDH) level, extra?age >60 years, elevated LDH level, extra?nasal primary site, and regional lymph node involvement were significantly associated with lower 5?year PFS rate. The nomogram included the primary site and regional lymph node involve?ment based on multivariate analysis. The calibration curve showed good agreement between the predicted and actual 5?year OS and PFS rates, and the C?indexes of the nomogram for the OS and PFS rates were 0.697 and 0.634, respectively. Conclusions: The primary site and regional lymph node involvement are independent prognostic factors for early?stage ENKTCL treated with chemotherapy followed by definitive radiotherapy.

OBJECTIVE：To establish a method for the content determination of potential genotoxic impurity maleic hydrazide in azintamide raw material. METHODS ：HPLC-FLD method was adopted. The determination was performed on Thermo Syncronis C18 column with mobile phase consisted of 0.2 mol/L acetic acid-methanol （gradient elution ）. The column temperature was set at 30 ℃，the excitation wavelength was 315 nm and emission wavelength was 389 nm. The flow rate was 1 mL/min，and the sample size was 20 μL. RESULTS：The blank solvent and azintamide did not interfere with the determination of maleic hydrazide. The linear range of maleic hydrazide was 19.5-300 ng/mL（r＝0.999 9）. The limit of detection was 4.5 ng/mL and the limit of quantification was 19.5 ng/mL. The recovery ranged from 98.79％ to 103.76％（RSDs were lower than 3.00％，n＝9）. RSDs of precision and stability （24 h）tests were no more than 5.63％，and those of durability tests were less than 2.00％（n＝6）. Maleic hydrazide was not detected in 3 batches of azinamide raw material. CONCLUSIONS ：The method is specific ，sensitive and accurate. It can be used for the trace determination of maleic hydrazide in azintamide or other matrix.

OBJECTIVES: To investigate whether necrostatin-1 (Nec-1) can protect islet cells from the damage induced by TNF-α. METHODS: After isolation and purification, the neonatal porcine islet cell clusters (NICCs) were divided into 3 groups (islets 10 000 IEQ/group): a Nec-1 group (Nec-1+TNF-α was added to the culture medium), a TNF-α group (TNF-α was added to the culture medium), and a control group (pure medium). The number of cells was observed after 48 h of co-culture. The cell death was evaluated by AO/EB staining. Insulin secretion and DNA of islets were detected by chemiluminescence and nucleic acid quantitative analysis. RT-PCR assay was used to examine the mRNA expressions of insulin gene, glueogan gene and somatostatin gene. Flow cytometry analysis was used to detect the viability of B cells. RESULTS: The number of islets in Nec-1 group, TNF-α group and the control group were (8 425±2 187), (4 325±778), and (7 122±1 558) IEQ, respectively. Compared to the other two groups, the number of dead cells in TNF-α group was greatly increased. The insulin/DNA values in the Nec-1 group, TNF-α group and blank control group were (13.21±3.15), (2.47±0.45), and (7.44±0.97) mIU/mg, respectively. Compared to the TNF-α group and the control group, the mRNA relative expression levels of insulin gene (6.73±1.07), glucagon gene (10.13±1.98), somatostatin gene (8.57±1.11) were significantly increased in the Nec-1 group (all <0.05), the rate of live cells (97.32±1.87)% and live B cells (90.86±3.68)% were increased significantly in the Nec-1 group (all <0.05). CONCLUSIONS TNF-α can induce neonatal porcine islet cells damage, which is attenuated in the presence of Nec-1. Nec-1 can increase the content of endocrine cells in NICCs.
Animals ; Imidazoles ; Indoles ; Insulin ; Islets of Langerhans ; Swine ; Tumor Necrosis Factor-alpha ; genetics

Objective:To analyze the clinical features and magnetic resonance imaging of non-malignant patients assigned to Prostate Imaging Reporting And Data System (PI-RADS) 5 score.Methods:We performed a retrospective review of 289 patients who underwent magnetic resonance ultrasound targeted combined system biopsy with PI-RADS 5 lesions in the First Affiliated Hospital of Nanjing Medical University between May 2019 and July 2021. The median age 72 (66, 77)years, median body mass index 24.4(22.3, 27.1)kg/m 2, median prostate volume (PV) 37.39(29.39, 48.86) ml, median PSA 22.24(10.91, 62.69) ng/ml, and median PSAD 0.53(0.30, 1.52)ng/ml 2 were recorded. According to the biopsy pathological results, all patients were divided into benign lesion group and prostate cancer group. PSA, PSAD, PV, and apparent diffusion coefficient (ADC) values were compared, and magnetic resonance imaging and clinical characteristics of patients with biopsy benign lesions were analyzed. Results:There were 11 cases (3.8%) with benign lesion and 278 cases (96.2%) with prostate cancer. The characters of 11 negative biopsy cases were displayed as follows: median age 69(66, 79)years, median body mass index 22.0(21.0, 25.5)kg/m 2, median PV 62.90(38.48, 71.96)ml, median PSA 5.55(2.99, 20.52)ng/ml, median PSAD 0.16(0.07, 0.24) ng/ml 2, median ADC 714.47(701.91, 801.26)×10 -6 mm 2/s, abnormal digital rectal and amination in 5 cases, smoking in 7 cases, and alcohol consumption in 4 cases. The median PV [62.90(38.48, 71.96) vs. 37.21(29.22, 47.82)ml, P<0.01], the PSA value [5.55(2.99, 20.52) vs. 23.53(11.14, 65.98)ng/ml, P<0.01], and the PSAD value [0.16(0.07, 0.24) vs. 0.58(0.31, 1.57)ng/ml 2, P<0.01] were significantly different between benign condition group and prostate carcinoma group. Benign condition group included 5 chronic prostatitis, 2 acute prostatitis (1 with focal adenocarcinoma), 2 granulomatous inflammation, and 2 tuberculous granulomatous inflammation. In 7 benign cases, PSA was less than 10 ng/ml, combined with frequent urination, urgency of urination and incontinence were founded. In 8 benign cases, the area of lesion was more than 50% of the total prostate area in the axial position and the imaging of magnetic resonance were diffused, with regular shape and uniform signal. The imaging of symmetrical distribution was in 6 cases. Conclusions:The benign condition with PI-RADS 5 lesions included chronic prostatitis, acute prostatitis, granulomatous inflammation and tuberculous granulomatous inflammation, among which prostatitis was the most common cause. The PSA value were less than 10 ng/ml in most benign cases, with symptoms such as frequent urination, urgency of urination and incontinence. The imaging of magnetic resonance were diffused, symmetrically distributed, with regular shape and uniform signal.

Objective:To establish and internally validate a nomogram model for predicting complicated acute appendicitis (CA).Methods:The clinical data from 663 acute appendicitis patients from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from October 2015 to October 2022 were retrospectively analyzed. There were 411 males and 252 females, aged ( M (IQR)) 41 (22) years (range: 18 to 84 years). There were 516 cases of CA and 147 cases of uncomplicated acute appendicitis. The minimum absolute contraction and selection operator regression model was used to screen the potential relative factors of CA, and the screened factors were included in the Logistic regression model for multivariate analysis. Software R was used to establish a preoperative CA nomogram prediction model, the receiver operating characteristic curve of the model was drawn, and the value of area under the curve (AUC) was compared to evaluate its identification ability, and the Bootstrap method was used for internal verification. Results:The elderly (age≥60 years) ( OR=2.428, 95% CI: 1.295 to 4.549), abdominal pain time (every rise of 1 hour) ( OR=1.089, 95% CI: 1.072 to 1.107), high fever (body temperature≥39 ℃) ( OR=1.122, 95% CI: 1.078 to 1.168), total bilirubin (every rise of 1 μmol/L) ( OR=2.629, 95% CI: 1.227 to 5.635) were independent relative factors of CA (all P<0.05). The AUC of this model was 0.935 (95% CI: 0.915 to 0.956). After internal verification using the Bootstrap method, the model still had a high discrimination ability (AUC=0.933), and the predicted CA curve was still in good agreement with the actual clinical CA curve. Conclusion:The clinical prediction model based on the elderly (age≥60 years), prolonged abdominal pain time, high fever (body temperature≥39 ℃), and increased total bilirubin can help clinicians effectively identify CA.