Objective To look for a method of LC with minimal invasion and better exposure. Methods During laparoscopic cholecystectomy with three holes, a needle with a No.7 thread was pierced to the abdominal cavity at the right eighth intercostal space and the fundus of the gallbladder was sutured. The gallbladder traction was performed by the thread to help exposure of the operative filed. LC of 90 cases were operated on by the method. Results All of the 90 cases were cured without any complication. Conclusions External abdominal gallbladder traction is more effective with minimal invision and better exposure as compared with the routine LC.

Ojective To study the method of laparoscopic fenestration and its effect for the treatment of hepatic and renal cysts and polycystic livers and kidneys. Methods Laparoscopic fenestration of the cysts was performed Results All of the 15 cases were operated on successfully and recovered smoothly. Conclusions laparoscopic fenestration of hepatic and renal cysts and polycystic livers and kidneys has all the advantages of minimally invasive surgery. Relapsed cysts can be also treated by laparoscopic fenestration.

Objective To study the causes and prevention of complications during LC. Methods The clinical data of 1200 patients treated by LC in the latest 10 years were reviewed retrospectively. Results All of the 1200 cases recovered smoothly without severe complications. 18 cases were converted to open-cholecystectomy. Conclutions Complications of LC can be prevented by strict training, serious operative procedures, and mastery of causes and prevention of complications.

Tumors represent one of the primary threats to human life, with the dissemination of malignant tumors being a leading cause of mortality among cancer patients. Early diagnosis of tumors can reliably predict their progression, significantly reducing mortality rates. Tumor markers, including circulating tumor cells, exosomes, proteins, circulating tumor DNA, miRNAs and so on, generated during the tumor development process, have emerged as effective approach for early tumor diagnosis. Several methods are currently employed to detect tumor markers, such as polymerase chain reaction, Northern blotting, next-generation sequencing, flow cytometry, and enzyme-linked immunosorbent assay. However, these methods often suffer from time-consuming process, high costs, low sensitivity, and the requirement for specialized personnel. Therefore, a new rapid, sensitive, and specific tumor detection method is urgently needed.The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system, originating from the adaptive immune system of bacteria, has found extensive applications in gene editing and nucleic acid detection. Based on the structure and function of Cas proteins, the CRISPR/Cas system can be classified into two classes and six types. Class I systems consist of multiple Cas protein complexes, including types I, III, and IV, while Class II systems comprise single, multi-domain Cas proteins mediated by RNA, including types II (Cas9), V (Cas12), and VI (Cas13). Class II systems have been widely employed in the fields of biotechnology and nucleic acid diagnostics due to their efficient target binding and programmable RNA specificity. Currently, fluorescence method is the most common signal output technique in CRISPR/Cas-based biosensors. However, this method often requires the integration of signal amplification technologies to enhance sensitivity and involves expensive and complex fluorescence detectors. To enhance the detection performance of CRISPR/Cas-based biosensors, the integration of CRISPR/Cas with some alternative techniques can be considered. The CRISPR/Cas integrated electrochemical sensor (E-CRISPR) possesses advantages such as miniaturization, high sensitivity, high specificity, and fast response speed.E-CRISPR can convert the reactions between biomolecules and detecting components into electrical signals, rendering the detection signals more easily readable and reducing the impact of background values. Therefore,E-CRISPR enhances the accuracy of detection results. E-CRISPR has been applied in various fields, including medical and health, environmental monitoring, and food safety. Furthermore, E-CRISPR holds tremendous potential for advancing the detection levels of tumor markers.Among all types of Cas enzymes, the three most widely applied are Cas9, Cas12, and Cas13, along with their respective subtypes. In this work, we provided a brief overview of the principles and characteristics of Class II CRISPR/Cas single-effector proteins. This paper focused on the various detection technologies based on E-CRISPR technique, including electrochemical impedance spectroscopy, voltammetry, photoelectrochemistry, and electrochemiluminescence. We also emphasized the applications of E-CRISPR in the field of tumor diagnosis, which mainly encompasses the detection of three typical tumor markers (ctDNA, miRNA, and proteins). Finally, we discussed the advantages and limitations of E-CRISPR, current challenges, and future development prospects. In summary, althoughE-CRISPR platform has made significant strides in tumor detection, certain challenges still need to be overcome for their widespread clinical application. Continuous optimization of the E-CRISPR platform holds the promise of achieving more accurate tumor subtyping diagnoses in clinical settings, which would be of significant importance for early patient diagnosis and prognosis assessment.

Objective To explore the value of time-resolved angiography with interleaved stochastic trajectories (TWIST)in providing hemodynamic indices and morphological imaging of calf arteries in patients with peripheral arterial occlusive disease(PAOD) with 3.0 T MR scanner. Methods Forty patients with confirmed or suspected PAOD underwent TWIST MRA for the calf arteries and conventional contrast-enhanced MRA(CE-MRA) for the whole lower peripheral arteries. TWIST MRA data were used to determine the bolus arriving time of the popliteal artery, the time difference of the bolus arrival in the popliteal artery between the two legs, calf artery transit time, and the mean peak enhancement time of calf arteries. The mean value of peak enhancement time of calf arteries was correlated with ankle-brachial index(ABI). The calf arteries were divided into 13 segments. The visible score of arterial segment was recorded, and the degree of arterial stenosis was graded too. In 16 patients, DSA was used as the gold standard to evaluate the sensitivity and specificity of TWIST MRA and CE-MRA. Results The bolus arriving time of the popliteal artery was (0.7±7.6) s the time difference of the bolus arrival in the popliteal artery between the two legs was (2.1±2.5) s, the calf artery transit time was (35.6±16.9) s, the mean value of peak enhancement time of calf arteries was (52.6±17.6) s. ABI was acquired from 24 patients and 48 legs, which correlated well with the mean value of peak enhancement time in calf arteries (r＝0.627,P＜0.01). The mean visible score was (2.83±0.48) of TWIST MRA images and (2.78±0.43) of CE-MRA images. There was no significant difference between them (P＞0.05). On CE-MRA, there were 38 legs with varying degrees of early venous enhancement; in contrast, there was no venous contamination on TWIST MRA. The sensitivity and specificity of TWIST MRA were 96.7% (118/122) and 100.0% (72/72) in showing normal artery and mild stenosis, 94.1% (32/34) and 96.2% (154/160) in showing severe stenosis, and 94.7% (36/38) and 98.7% (154/156) in showing occlusion. The sensitivity and specificity of CE-MRA were 95.9% (117/122) and 100.0% (72/72) in showing normal artery and mild stenosis, 91.2% (31/34) and 95.6% (153/160) in showing severe stenosis, and 94.7% (36/38) and 98.1% (153/156) in showing occlusion. Conclusion TWIST MRA is a preferable choice for calf arterial occlusive disease, which provided nice morphological images and useful hemodynamic indices with simple operation.

Objective To observe the changes of urine microalbumin (mAlb), renal function and intima-media thickness(IMT) of the carotid artery in prehypertensive patients and investigate whether there are related target organ damage. Methods From August 2009 to May 2010,one hundred and eighty patients were recruited into the study and divided into 3 groups according to the 2005 revised edition of guideline for prevention and treatment of hypertension in China: Sixty normotensive subjects ( NT group,41 males, 19 females,mean age [54. 6 ± 12. 3] years old) ;Sixty prehypertensive patients( PH group,32 males,28 females,mean age [57.0 ± 12.7] years old) and 60 hypertensive patients ( EH group, 28 males, 32 females, mean age [65.0 ±12. 1] years old). Urine mAlb, serum creatinine(Cr) and IMT of the carotid artery were measured and compared among the three groups. The relationships between prehypertension and urine mAlb,serum Cr, IMT of the carotid artery were analyzed. Results In the NT group, PH group and EH group, the urine mAlb concentrations were ( 15.3 ± 7.5 ) mg/L, ( 24. 6 ± 10. 7 ) mg/L and ( 37.3 ± 20. 4) mg/L respectively ;serum Cr were ( 68.0 ± 16. 5 )μmol/L, (81.9 ± 14.9)μmol/L and (95.8 ± 22.5)μmol/L respectively; IMT of the carotid artery were ( 1.46 ± 0. 67 ) mm, ( 1.79 ± 0. 74 ) mm and( 2. 34 ± 0. 78 ) mm respectively. Urine mAlb, serum Cr and IMT of the carotid artery were significantly higher in the PH group and EH group when compared to NT group ( P ＜0. 05 or 0. 01 ); and in the EH group these indices were also significantly higher than those in the PH group( P ＜0. 01 ). Multiple linear regression analysis showed that after adjusted age, body mass index (BMI), blood pressure( BP), serum lipid and serum glucose, in the PH group,the standard partial regression coefficients of correlation between urine mAlb and systolic blood pressure (SBP), diastolic blood pressure(DBP) and serum glucose were 0. 279,0. 259 and 0. 360 respectively ( P = 0. 012, 0. 043 and ＜ 0. 001 ); the standard partial regression coefficients of correlation between serum Cr and SBP, DBP and serum glucose were 0. 317,0. 257 and 0. 377 respectively ( P = 0. 006,0. 049 and ＜ 0. 001 ). The standard partial regression coefficients of correlation between IMT of the carotid artery and age, SBP, DBP, total cholesterol and serum glucose were 0. 381,0. 375,0. 263,0. 265 and 0. 372 respectively( P ＜ 0. 001, ＜ 0. 001, = 0. 033,0. 021 and ＜ 0. 001 ). In the EH group, the standard partial regression coefficients of correlation between urine mAlb and SBP, DBP and serum glucose were 0. 378,0. 258 and 0. 283 respectively( P ＜ 0. 001, = 0. 046, and 0. 009); the standard partial regression coefficients of correlation between serum Cr and SBP, DBP and serum glucose were 0. 294,0. 261 and 0. 464respectively (P =0. 008,0. 042 and ＜0. 001 ) ;the standard partial regression coefficients of correlation between IMT of the carotid artery and age, SBP, high density lipoprotein cholesterol and serum glucose were 0. 262,0. 264, -0. 374 and 0. 306 respectively ( P = 0. 035,0. 023, ＜ 0. 001 and 0. 007 ). We found that both urine mAlb and serum Cr were significantly associated with BP and serum glucose; IMT of the carotid artery was significantly associated with age, BP, serum lipid and serum glucose. Conclusion Urine mAlb, serum Cr andIMT of the carotid artery increased in prehypertensive patients, and these indices were significantly associatedwith the level of the BP. These data indicated there were related target organ damage occurred in prehypertensive patients,the BP level was the risk factor that increased urine mAlb,serum Cr and IMT of the carotid artery.

Objective To investigate the association between morniing blood pressure surge (MBPS) and carotid atherosclerosis in elder patients with essential hypertension. Methods According to the results of24h ambulatory blood pressure monitoring, 106 patients were classified as the morning BP surge group (MBPS group,n = 58) ,and nonsurge group (NMBPS group, n = 48). Patients underwent carotid ultrasound and the intima-medial thickness (CCA-IMT) and plaques were examined. Results The CCA-IMT of the MBPS group was significantly thicker than that the NMBPS group[(1.27 ± 0. 12)mm vs (0.92 ± 0.33 )mm], P ＜ 0. 05 ) ;②Compared with the NMBPS group,the severity of carotid arteries plaque of the MBPS group was significantly higher (72. 15% vs 54.21% ), ( P ＜0. 01 ) ;③Pearson relation analysis showed CCA-IMT level positively correlated with age (r = 0.288, P ＜ 0.001 ) ,the average of 24h SBP ( r = 0. 768 ,P ＜ 0. 001 ), and MBPS ( r = 0. 768, P ＜ 0.001 ). Conclusion The study showed that MBPS was closely related with carotid atherosclerosis in elder patients with essential hypertension and was an important risk factor in the process of atheresclerosis.

AIM: To study the protective effects of HIF-1α gene transfection on hypoxic injury in human HepG2 cells. METHODS: After gene transfection, HepG2 cells were randomly divided into 4 groups: normoxia with Ad-GFP transfected group, normoxia with Ad-HIF-1 transfected group, hypoxia with Ad-GFP transfected group and hypoxia with Ad-HIF-1 transfected group. LDH leaking rate, cell viability, contents of NO and ROS, the iNOS activity were measured. RESULTS: High levels of HIF-1α mRNA and protein were detected in Ad-HIF transfected HepG2 cells. Cell viability was significantly lower in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05). No marked difference of cell viability was found between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. ROS was significantly higher in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05), while no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group or between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group. The content of NO and iNOS activity were significantly higher in Ad-HIF transfected-normoxia group and Ad-GFP transfected-hypoxia group than those in Ad-GFP transfected-normoxia group (P<0.05), no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group or between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. CONCLUSION: Higher HIF-1α expression is contributed to protective effects against hypoxic injury in HepG2 cells, the mechanisms of which may be correlated with promoting expression of gene regulated by HIF-1 and restraining over-expression of injure factors.

Objective To investigate the clinical features of invasive bronchial-pulmonary aspergillosis (IBPA) in critically ill patients with chronic obstructive pulmonary diseases (COPD).Methods Clinical data of 7 COPD patients complicated with IBPA from respiratory intensive care unit (RICU) in Beijing Chaoyang Hospital during April 2006 and August 2012 were retrospectively analyzed.Results All patients were treated with several kinds of broad-spectrum antibiotics and 6 patients were also treated with large dose of corticosteroids before their admission to RICU.The most frequent clinical manifestations were fever,persistent bronchospasm induced dyspnea and bilateral wheezing which were unresponsive to large doses of corticosteroids and broad-spectrum antibiotics.In 5 patients with both aspergillus tracheobronchitis (ATB) and invasive pulmonary aspergillosis (IPA),bronchoscopy showed tracheobronchial inflammation,bronchospasm,phlegm and bronchial pseudomembrane formation.All these 5 patients had a rapid progression from normal to infiltrates or consolidation in chest X-ray,and died of respiratory failure or multiple organ failure.The other 2 cases were diagnosed with ATB and received prophylactic antifungal treatments before infiltrates were shown on their chest X-ray.Both of them survived.Conclusions In COPD patients combined with IBPA,ATB may progress rapidly to IPA which affect lung parenchyma and lead a high mortality.Bronchoscopy and bronchial mucous biopsy may be helpful in early diagnosis and treatment.

ObjectiveTo explore the pharmacodynamic ingredients of Zhenqi Fuzheng granules （ZFG） for immunomodulatory through spectrum-effect relationship analysis， which provides experimental basis for improving the quality standard of ZFG. MethodEighteen batches of ZFG from six manufacturers were collected for analysis. The fingerprints were established by high performance liquid chromatography （HPLC）. Acetonitrile （A）-0.1% formic acid aqueous solution （B） were adopted as the mobile phase with gradient elution （0-15 min， 5%A； 15-23 min， 5%-8%A； 23-30 min， 8%-11%A； 30-45 min， 11%-18%A； 45-60 min， 18%-21%A； 60-67 min， 21%-23%A； 67-90 min， 23%-37%A）， the detection wavelength was 220 nm. Chemometric analysis such as similarity analysis and hierarchical cluster analysis （HCA） were subsequently used to analyze the similarities and chemical differences among these samples. A cyclophosphamide-induced immunodeficiency mouse model was used to evaluate the immune-enhancing effects of the products from different manufacturers. The spectrum-effect relationship between HPLC fingerprints and the immunomodulatory effects was examined using Spearman bivariate correlation analysis. HPLC coupled with mass spectrometry （HPLC-MSⁿ） was used to identify the spectrum-effect related peaks with electrospray ionization， positive and negative ion modes， and scanning range of m/z 100-1 500. ResultThe HPLC fingerprint of ZFG was established， and twenty peaks with good resolution were selected as common peaks. The results of quality analysis and pharmacodynamic test showed there were significant differences in both ingredients content and immune-enhancing effects of ZFG from different manufacturers. Through spectrum-effect relationship study， twelve peaks were screened as bioactive ingredients peaks. Thereafter， eight peaks among them were subsequently identified by HPLC-MSⁿ. They were salidroside （peak 2）， echinacoside （peak 5）， calycosin-7-glucoside （peak 6）， isomer of specnuezhenide （peak 7）， isonuezhenide （peak 9）， calycosin （peak 11）， nuezhenide G₁₃ or oleonuezhenide （peak 14）， and formononetin （peak 18）， respectively. ConclusionThere are differences in quality and efficacy of ZFG produced by different manufacturers. Through spectrum-effect relationship analysis， the medicinal ingredients of ZFG for immune-enhancing effects are screened， which can provide reference for the improvement of its quality standard.