Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10%FBS, during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation, including three-drug combination group and four-drug combination group. The protocol of mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination induced differentiation, four-drug combination was successfully achieved in over 90% of the cell inducing, which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.

Objective To investigate the relationship between pleural resonance features and the chest volume and vital capacity in young male adults .Methods A total of 60 healthy young male adults were included in this study .Energy distribution of pleural resonance was measured with the multi-channel voice analysis system when they pronounced /a:/.The frequency spectrums were 0~999 Hz (FR1 ) ,1 000~1 999 Hz (FR2 ) ,2 000~2 999 Hz (FR3 ) ,3 000~4 000 Hz (FR4 ) .Fast 3D reconstruction of chest was detected by multi slice spiral CT to calcu‐late the chest volume .The vital capacity was evaluated by aerodynamics system .SPSS 18 .0 software was used to analyze the data .Results The vital capacity in healthy young male adults was 4 .31 ± 0 .63 L ,the chest volume was 5 .69 ± 0 .52 L .The frequency spectrum of FR1 was 53 .38% ± 2 .14% ,FR2 was 30 .72% ± 1 .59% ,FR3 was 10 . 53% ± 2 .75% ,FR4 was 5 .35% ± 2 .32% ,respectively .There was a highly positive correlation between the chest volume and FR1 (r=0 .854) ,moderately positive correlation between the volume and FR2 (r=0 .740) ,moderately negative correlation between the volume and FR3 (r= -0 .587) ,moderately negative correlation between the volume and FR4 (r= -0 .565);There was a highly positive correlation between the vital capacity and FR1 (r=0 .744) ,mod‐erately positive correlation between the volume and FR2 (r=0 .699) ,moderately negative correlation between thevolume and FR3 (r= -0 .632) ,weakly negative correlation between the volume and FR4 (r= -0 .429) .Conclusion There was a high correlation between the chest volume and pleural resonance ,high correlation between vital capacity and pleural resonance in young male adults ,which may be a influencing factor of pronunciation effect .

Objective: To investigate the effect of electroacupuncture (EA) on rat with diet induced obesity (DIO) and to explore the possible neurochemical mechanisms using the technique of immumohistochemisty. Methods:To establish DIO rat model by feeding the animals with high fat diet for 14 weeks. DIO rats were randomly divided into 4 groups: (1) 2Hz EA group, (2) 100Hz EA group, (3) restrain control group,(4) diet resistance (DR) group,(5) DIO group and (6) normal control group. EA treatment: (1) The acupoints used were Zusanli and Sanyinjiao on both legs. (2) The intensities of stimulation were 0.5, 1.0 and 1.5mA for 10 mins each. EA treatment was administered 3 times per week. Food intake and body weight were measured daily for 4 weeks. (3) The changes of the expression of ? melanocyte stimulating hormone (? MSH) in the hypothalamic arcuate nucleus (ARC) were measured with immunohistochemical semiquantitative analysis. Results: (1) The food intake and body weight of 2 Hz EA group and 100 Hz EA group were decreased significantly compared with the restrain control group and DIO group. (2) The number of ? MSH positive cells in hypothalamic ARC in 2 Hz EA and 100 Hz EA group was significantly higher than that in restrain control group and DIO group. The number of ? MSH positive cells in hypothalamic ARC in DIO group is significantly lower than those in DR group or normal control group. Conclusion: A decrease of ? MSH level in hypothalamus may be associated with diet induced obesity. The therapeutic effect on obesity produced by EA may be accounted for by the stimulation of pro opio melanocortin neurons in hypothalamic ARC to release ? MSH, which inhibits food intake , resulting in a decrease of body weight.

OBJECTIVESTo study the shape and the structures in the craniocervical junction region (CCJR) and the safety of far lateral approach.

METHODSTen cadaveric heads and 20 skull-base specimens were fixed with 10% formalin, and 4 of the 10 cadaveric heads were sectioned in different plane with the aid of operative microscope. The shape and relation of structures in the CCJR were observed.

RESULTSThe third segment of the vertebral artery(VA) was 21.6(15.3 31.9) mm. An incomplete bony canal was found to be 15% and complete bony canal surrounding the VA to be 5% on the VA groove. The distance of the left and right VA was 14.3a(c)(9.8 15.2) mm on the entry into the dura. The length of the occipital condyle was 26.8(25.1 28.2) mm, with the thickness of its anterior, middle and posterior one-third part was 9.9(9.6 10.6) mm, 11.2(9.2 13.1) mm and 8.6 (8.3 9.0) mm respectively. The distance between the posterior pole of the occipital condyle and the intracranial orifice of the hypoglossal canal was 9.9(8.6 11.4) mm, and between the posterior pole and the extracranial orifice was 16.1(13.5 17.1) mm. The transverse process of the atlas was the most important bony landmark for the approach.

CONCLUSIONSKnowing the shape and relationship of the VA and occipital condyle in the CCJR is helpful in preserving the important structures in far lateral approach to target region.

Humans ; Microsurgery ; Occipital Bone ; anatomy & histology ; Skull ; anatomy & histology ; surgery ; Vertebral Artery ; anatomy & histology

OBJECTIVESTo explore shape and position relationship of the structures with in sinus triangle region, and provide the anatomic foundation for extended presigmoid operative approach.

METHODSTen cadaveric heads and 15 skull-base specimens fixed with 10% formalin, with aid of operative microscope, the shape and relations of structures with in the sinus triangle region were observed and microphotographed. Four cadaveric heads were sectioned in different plane using collodion embedding process.

RESULTSJugular blub (JB) had high -placed jugular blub (HJB), normal jugular blub (NJB) and low-placed jugular blub (LJB). The incidence of HJB was 8% and 18% for the left and right sides (P < 0.01). The JB dome can extend upward the hypotympanic cavity and the posterior wall of the internal acoustic meatus. The incidence of NJB and LJB was 36% and 30%, and 6% and 2% respectively. The level distance between the posterior semicircular canal and petrosal posterior surface was (4.1 +/- 1.1) mm. The distance between the JB and vertical part of the facial nerve was (3.2 +/- 0.5) mm and (2.0 +/- 0.4) mm. The distance between the JB and posterior margin of the internal acoustic port was (7.5 +/- 1.7) mm and (4.0 +/- 1.0) mm.

CONCLUSIONSKnowing the shape and position relationship of structures in the sinus triangle region is favorable for preserving the important structures in extended presigmoid operative approach.

Autopsy ; Cadaver ; Cranial Sinuses ; anatomy & histology ; Glomus Jugulare ; anatomy & histology ; Humans ; Neuroanatomy ; Semicircular Canals ; anatomy & histology

Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.