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 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

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

Objective:To assess the rationality of the maximum lesion diameter of 15 mm in prostate imaging reporting and data system(PI-RADS)as a criterion for upgrading a lesion from category 4 to 5 and improve it to enhance the prediction of clinically significant prostate cancer(csPCa).Methods:In this study,the patients who underwent prostate magnetic resonance imaging(MRI)and prostate biopsy at Peking University First Hospital from 2019 to 2022 as a development cohort,and the patients in 2023 as a validation cohort were reviewed.The localization and maximum diameter of the lesion were fully evalua-ted.The area under the curve(AUC)and the cut-off value of the maximum diameter of the lesion to pre-dict the detection of csPCa were calculated from the receiver operating characteristics(ROC)curve.Confounding factors were reduced by propensity score matching(PSM).Diagnostic efficacy was com-pared in the validation cohort.Results:Of the 589 patients in the development cohort,358(60.8％)lesions were located in the peripheral zone and 231(39.2％)were located in the transition zone,and 496(84.2％)patients detected csPCa.The median diameter of the lesions in the peripheral zone was smaller than that in the transition zone(14 mm vs.19 mm,P＜0.001).In the ROC analysis of the maximal diameter on the csPCa prediction,there was no statistically significant difference between the peri-pheral zone(AUC=0.709)and the transition zone(AUC=0.673,P=0.585),and the cut-off values were calculated to be 11.5 mm for the peripheral zone and 16.5 mm for the migrating zone.By calcula-ting the Youden index for the cut-off values in the validation cohort,we found that the categorisation by lesion location led to better predictive results.Finally,the net reclassification index(NRI)was 0.170.Conclusion:15 mm as a criterion for upgrading the PI-RADS score from 4 to 5 is reasonable but too general.The cut-off value for peripheral zone lesions is smaller than that in transitional zone.In the future consideration could be given to setting separate cut-off values for lesions in different locations.

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.

Objective:To investigate the diagnostic efficacy of targeted biopsy combined with regional systematic biopsy in prostate cancer(PCa)in patients with prostate imaging reporting and data system v2.1(PI-RADS v2.1)4-5.Methods:From January 2023 to October 2023,patients who underwent prostate biopsy for the first time with total prostate specific antigen(tPSA)≤20 ng/mL and had a multi-parametric magnetic resonance imaging(mpMRI)PI-RADS of 4-5 in Peking University First Hospital were prospectively collected.All the patients underwent transrectal ultrasound-guided cognitive fusion tar-geted biopsy(3 cores)followed by systematic biopsy(12 cores).Various hypothetical biopsy schemes were defined based on different biopsy sites.The detection effectiveness of targeted biopsy combined with regional systematic biopsy and other biopsy schemes for prostate cancer were compared using Cochran's Q and McNemar tests.Results:A total of 255 patients were enrolled,of whom 204(80.0％)were de-tected with prostate adenocarcinoma and 187(73.3％)were clinically significant with prostate cancer(csPCa).The detection rate of PCa with targeted biopsy was significantly lower than that of targeted biopsy combined with 12-core system biopsy(77.3％vs.80.0％,P=0.016),and 71.4％(5/7)of the missed patients was csPCa.There was no significant difference in the detection rate between targeted biopsy combined with 4-core regional system biopsy and 12-core system biopsy(P＞0.999),and 1 case of csPCa and clinically insignificant prostate cancer(cisPCa)were missed.There was no significant difference in the detection rate of PCa between targeted combined regional system biopsy and targeted combined lateral or traditional 6-core system biopsy and the number of cores were reduced.Missed diag-nosis of targeted biopsy was correlated with the maximum diameter of the lesion(OR=0.086,95％CI:0.013-0.562,P=0.010).For the patients with PI-RADS 5,only 1 case of PCa was missed in 122 cases by targeted biopsy alone.For patients with PI-RADS 4,6 PCa cases were missed among the 133 patients with targeted biopsy alone,and 1 case of csPCa and cisPCa were missed by targeted biopsy com-bined with regional system biopsy.The statistics of positive core counts for different biopsy schemes indi-cated that targeted combined regional systematic biopsy had a higher proportion of positive cores second only to targeted biopsy alone.Conclusion:Targeted biopsy combined with regional systematic biopsy has high diagnostic efficacy in patients with PI-RADS 4-5 and can be considered as one of the improved schemes for combined biopsy.Targeted biopsy alone is also a feasible option for patients for patients with a PI-RADS score of 5.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.