A 68-year-old patient with non-small cell lung squamous carcinoma who received 6 cycles of sintilimab combination chemotherapy and sintilimab 200 mg,ivd,monotherapy developed severe diarrhea,abdominal pain,blood in the stool and other discomforts,and ultrasound and colonoscopy demonstrated extensive congestion and inflammation in the intestinal tract,and the pathologic biopsy was comprehensively considered to be an acute immune enteritis.Immunotherapy was suspended,adequate glucocorticoids and symptomatic treatment were given,and the patient's diarrhea and blood in stool improved after 2 days,and the symptoms were relieved and returned to normal after 6 days.The association between the patient's immune enteritis and sintilimab was assessed as probably relerant.This article reviews the literature on the case of immune-associated enteritis caused by sintilimab,describes how to use experimental methods and enteroscopy to detect the pathological changes in the clinic;and combines them with the clinical manifestations of diarrhea and blood in the stools to make the diagnosis and differentiation;and then refers to the guideline grading for timely management;and discusses the case to improve the clinicians'ability to recognize and deal with the relevant scenarios.

The transcription factors (TFs) in the NAC family are involved in regulating multiple biological processes, playing an important role in plant growth, development, and stress adaptation. Our previous studies have demonstrated that TaNAC14, a member of the NAC family in wheat (Triticum aestivum L.), positively regulates root growth and development and enhances the drought tolerance of wheat seedlings. In this study, we analyzed the physicochemical properties and structure and verified the subcellular localization and transcriptional activation activity of TaNAC14. The prokaryotic expression vector pET21a-HMT-TaNAC14 was constructed and transformed into Escherichia coli BL21 CodonPlus (DE3)-RIPL. The conditions for inducing the expression of the recombinant protein HMT-TaNAC14 were optimized. The solubility of the recombinant protein was analyzed, and the protein was purified by affinity chromatography on a Ni-nitrilotriacetic acid column. The results indicated that TaNAC14 had a conserved domain of the NAM family. It was located in the nucleus and had transcriptional activation activity. The optimal conditions for expression of the recombinant protein in E. coli were induction with 0.2mmol/L IPTG for 4 h. The recombinant protein mainly existed in the soluble form, and the target protein was obtained after purification. This study lays a foundation for the identification of target genes regulated by TaNAC14.
Triticum/metabolism* ; Escherichia coli/metabolism* ; Plant Proteins/metabolism* ; Transcription Factors/metabolism* ; Recombinant Proteins/metabolism* ; Genetic Vectors/genetics*

【Objective】 To evaluate the efficacy and safety of different doses and frequencies of oral Sildenafil in the treatment of erectile dysfunction (ED). 【Methods】 The randomized,open clinical trial included 120 ED patients who met the inclusion and exclusion criteria. The patients were randomly divided into the following groups:on-schedule (25 mg/day),on-demand (50 mg,taken irregularly half an hour before each sexual life),new regular group (25 mg/day,50 mg more before each sexual life),regular group (100 mg/time,twice/week). All treatments lasted for 8 weeks. The follow-up indexes included the five-item International Index of Erectile Function (IIEF-5),Erection Hardness Scale (EHS) and Sexual Encounter Profile (SEP2/3). The adverse reactions were recorded. 【Results】 The IIEF-5 scores of the four groups were significantly higher than those after baseline treatment (P<0.001),but there was no significant difference among the four groups (P>0.05). In terms of effective rate,at the 16th week,there were significant differences between the on-demand group (10.7%) and new regular group (62.1%),and between the on-demand group (10.7%) and regular group (50.0%) (P<0.001). In terms of EHS, the percentage of grade 4 patients in regular group was significant higher than that in the on-demand group at the 8th week and 16th week (all P<0.05). In terms of positive rate of SEP-3,there was a significant difference between the on-demand group and regular group (P=0.042) at the 16th week. In the course of treatment,there were transient adverse reactions such as headache,blurred vision,stuffy nose and back pain,which did not affect the treatment. 【Conclusion】 All of the four treatment methods of oral sildenafil showed good efficacy. Both regular group and new regular group maintained good clinical efficacy during the follow-up,which is better than that of the on-demand group. The new regular scheme can be used as a new,safe and effective treatment option.

Objective To investigate the influence of overweight and obese on serum expressions of adipocytokines and prognosis of patients with acute leukemia. Methods Five hundred and seventy patients diagnosed as acute leukemia in the People's Hospital of Liaoning Province and Shengjing Hospital of China Medical University from July 2008 to July 2015 were collected. According to the body mass index (BMI), the patients were subdivided into two groups as obese/overweight group (BMI≥24 kg/m2) and control group (BMI<24 kg/m2). The avidinbiotincomplex-enzyme-linked immunosorbent assay (ELLSA) was used to detect the adiponectin, leptin, and resistin levels in the two groups. The t test was used to compare the difference of serum adiponectin, leptin and resistin expression between the two groups; Kaplan-Meier method was used to compare the difference of survival between the two groups. Results In patients with acute lymphoblastic leukemia (ALL)and acute myeloid leukemia (AML), the adiponectin levels in obese/overweight group were lower than those in the control group [adult ALL:(3.8±2.1) pg/ml vs. (6.4±2.9) pg/ml, child ALL:(4.2±2.7) pg/ml vs. (7.4±3.1) pg/ml, AML:(4.1±2.3) pg/ml vs. (6.9±3.1) pg/ml;t values were-2.291,-2.462,-2.244;P values were 0.023, 0.015, 0.026, respectively]. The leptin levels were high in the child ALL obese/overweight group than those in the control group (34±17 vs. 21±17) (t= 2.092, P= 0.038). The resistin levels of all the acuteleukemia patients did not have statistical difference between the obese/overweight group and control group(all P>0.05). In AML and adult ALL patients, the survival analysis showed that the 5-year overall survival rate in obese/overweight group were lower than that in the control group (38.0％vs. 46.6％,χ2= 1.449, P= 0.001;41.4％vs. 48.4％,χ2= 4.166, P= 0.041). Conclusion For the acute leukemia, the adiponectin levels are low in obese or overweight patients, and the 5-year survival rate of obese or overweight AML and adult ALL patients is lower than that of normal weight patients.

Objective To investigate the application and the effects of individualized hollow resin plugs in the prevention of external auditory canal stenosis after canaloplasty of external auditory meatus for microtia patients . Methods The clinical data of 65 patients (68 ears) with microtia and aural atresia hospitalized in our department from January ,2006 to December ,2015 were summarized .Otoplasty of external ears and canaloplasty of external auditory meatus were operated on all patients under general anesthesia .Thigh flap transplantation was lined in the bony canal .One month later ,individualized hollow resin plugs were made and worn for 6 to 12 months ,and fol-lowed up for 1～2 years .Results The shapes of the reconstructed auricles were satisfactory .Sixty-four ears with wide external auditory canal openings were recorded in 68 ears .Preoperative hearing was 70 .45 ± 5 .5 dB HL ,and postoperative hearing was 55 .55 ± 5 .2 dB HL .Their hearing was not affected .External auditory canal restenosis occurred in 4 ears .Conclusion The individualized hollow resin plugs do not affect the hearing in use ,and can effec-tively prevent the restenosis of the external auditory canal after microtia reconstruction .

Objective To investigate the effect of intense low frequency noise on the different hearing functions and hair cell damage in bama pigs.Methods Thirteen bama pigs were randomly divided into a normal control group (3 pigs) and an experimental group (10 pigs).The pigs in experimental group were randomly divided into 50 Hz subgroup (5 pigs) and 70 Hz subgroup (5 pigs).The 50 Hz subgroup was exposed to intense low frequency noise at 167～170 dB SPL,50 Hz for 30 min,and the 70 Hz subgroup was exposed to intense low frequency noise at 164 dB SPL,70 Hz for 30 min.Auditory brainstem response(ABR) and distortion product otoacoustic emission (DPOAE) were performed before and after noise exposure.The cochlear were collected and the inner ear morphology changes were studied.Results Before the experiment,the ABR threshold of the pig was 20～50 dB SPL,and the average of 50 Hz group was 33.5±9.4 dB SPL,the average of the 70 Hz group was 34.0±4.6 dB SPL.The level of the above 3 000 Hz DPOAE could be elicited.But they were not elicited after the exposure to the noise.The inner ear structures were damaged.The DAPI showed hair cell missing.Conclusion ABR and DPOAE were elevated after noise exposure.The auditory system of bama pigs had irreversible damages.Hair cells damaged were given priority to with necrosis,and the higher the level of intense low frequency noise,the more serious the damage to the hair cell.

BACKGROUND:The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention;however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.OBJECTIVE: To study the feasibility of cell-sheet ulturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct ell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to plantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after mplantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type Ⅱ collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited ature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the hondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.

Objective]To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.[Methods]rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. [Results]Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non-uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.[Conclusion]Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.

Objective: To observe the morphological changes on cochlear hair cells of rats in simulated weightlessness and inboard noise and to investigate the different changes in three turns of hair cells.Methods: Thirty-two healthy SD rats, all males, were randomly divided into four groups: control group, weightlessness group, noise group and weightlessness+noise groups (n=8).Then rats were exposed to-30° head down tilt as simulated weightlessness and inboard noise including steady-state noise which was (72±2) dB SPL and impulse noise up to 160 dB SPL in spaceship environment.The control group was kept in normal condition for 8 weeks.Bilateral auditory brainstem response (ABR) thresholds were tested before and after exposure respectively, and immunofluorescence staining and scanning electron microscopy (SEMs) of basilar membrane were applied after exposure.Results: ABR threshold shifts of each group were higher after exposure.There was difference between ABRs of the experiment groups before and after exposure (P<0.05).IF showed that the inner hair cells (IHCs) missing was the main damage in the basal turn of weightlessness group, the hair cells in the middle turn were swell and in the top turn, the hair cells were not clear.In noise group, the main loss happened in the outer hair cells (OHCs) of the outermost layer.In weightlessness+noise group, the nuclear missing in the basal turn was apparent, and mainly happened at the outermost layer.Meanwhile, the missing of hair cells in the middle turn and top turn was seen at the innermost layer.SEM showed that the cilia in the basal turn of weightlessness group were serious lodging, and occasional absence.Furthermore, the basal cilia in noise group became lodged and absent, and the other two turns were seriously missing.And in weightlessness+noise group, the cilia missing in the basal turn was apparently seen.The damage degree of the four groups: weightlessness+noise group>noise group>weightlessness group>control group and the damage degree of the four turns of hair cells: basal turn>mid turn>top turn.Conclusion: The rats exposed to the above environment for 2 weeks displayed obvious changes in cochlea morphology, and the weightlessness+noise group had the most obvious damage.

BACKGROUND:To seek for ideal scaffold materials is still an important task for cartilage tissue engineering.OBJECTIVE:To investigate the application of the AviteneTM microfibrillar collagen hemostat sponge in cartilage tissue engineering.METHODS:Rabbit auricular cartilage was harvested via surgical operation,and primary chondrocytes were isolated and amplified.Microfibrillar collagen hemostat sponge was cut into small bricks.The passage 2 chondrocytes were suspended and seeded onto the spongy bricks.After 1 week of in vitro culture,the constructs were then implanted into nude mice.After 8 weeks,the specimens were collected and evaluated using gross,histological and immunohistochamical observation.RESULTS AND CONCLUSION:During the cell seeding,the scaffold maintained its dimensions.No shrinkage was observed when the cell suspension was added.There was no considerable change in dimensions during the 1-week in vitro culture and at 8 weeks after implantation in nude mice.At 8 weeks post-implantation,mature cartilage blocks were harvested,which were white,translucent,and flexible.Histologically,the constructs appeared to have typical mature cartilaginous tissues,with robust extracellular matrix secretion,in which the microfibrillar collagen was incompletely degraded.We conclude that the microfibrillar collagen is a favorable scaffold material for cartilage tissue engineering.