Objective To determine the effects of individual antibiotic and immunosuppressive regime on postoperative infection in liver transplant recipients. Method There were 31 cases of liver transplantation from March 2001 to May 2005. The recipients received individual antibiotic and immunosuppressive regime based on the drug susceptibility testing and monitoring of blood drug concentration. The incidence and pattern of infection and the mortality in these recipients were analyzed retrospectively. Results There were 15 episodes of infection during recipients' staying in hospital. The common etiologies were Enterobacter cloacae, pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, and Staph. epidermidis. According to the drug sensitive test, targocid/tienam and tazocin were mostly used in antibiotic regime for treatment of postoperative infection. With monitoring of blood concentration, appropriate application of immunosuppressive agents decreased the incidence of infection from 86.7% before 2003 to 48.4% after 2003 (P0.05). Conclusion Individual application of antibiotic and immunosuppressive regime leads to the suppression of infections and other complications in liver transplant recipients.

Objective To evaluate the potential of interventional regional arterial infusion for severe acute pancreatitis (SAP). Methods Regional arterial infusion with somatostatin and antibiotic was performed in 20 cases of SAP within 4 hours after diagnosis confirmed,and pressurized infusion lasted for 5 days. Another 28 cases of SAP as control group received drug treatment all over the body. The SAP clinical data such as symptom,mortality,complication rate and stay time in hospital were analysed in two groups. Results There were significant differences in remission,serum amylase,urine amylase and lipase between the regional and total infusion groups. The stay time in hospital,mortality,complication and operation rate were lower in regional infusion group than those of control group(P

Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot.
OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s.
METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method.
RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.

Sorafenib, as the first-line treatment drug for advanced hepatocellular carcinoma (HCC), can effectively improve the prognosis of patients with HCC, but sorafenib resistance has become a major obstacle to the therapeutic outcome of HCC. Recent studies have shown that non-coding RNA plays a key role in sorafenib resistance in HCC. This article summarizes that non-coding RNA regulates the sensitivity of HCC to sorafenib by inducing the autophagy of hepatoma cells, promoting the proliferation of liver cancer stem cells, promoting the EMT process of hepatoma cells, inhibiting the apoptosis of hepatoma cells, and regulating the microenvironment of liver cancer tissue. It is pointed out that the molecular mechanism of non-coding RNA in regulating sorafenib resistance in HCC has potential clinical significance in overcoming sorafenib resistance in HCC.

Objective:To study the effect of diabetes mellitus (DM) on sepsis in patients with pyogenic liver abscess (PLA).Methods:The clinical data of 116 patients with PLA treated in the Affiliated Hospital of Jiangnan University from January 2021 to May 2022 were retrospectively analyzed, including 64 males and 52 females, aged (62.3±12.6) years old. Patients were divided into DM group ( n=56) and non-DM group ( n=60), which were also divided into the sepsis group ( n=29) and the non-sepsis group ( n=87). The clinical features were compared among the groups, the risk factors of PLA complicated with sepsis were analyzed by multivariate logistic regression. Mediation model was used to analyze how DM affects the development of sepsis. Results:Compared with the non-DM group, patients in DM group had higher incidences of hypertension and acute physiology and chronic health evaluation II, a higher proportion of blood neutrophil count, a higher serum levels of triglyceride, urea nitrogen, fasting blood glucose and glycated hemoglobin at admission. The DM group also higher incidences of hypoproteinemia, pleural effusion, and sepsis, with longer hospital stay and higher hospitalization cost (all P<0.05). The levels of hemoglobin, albumin and hematocrit were lower in DM group (all P<0.05). Multivariate logistic regression analysis showed that comorbidity of DM ( OR=3.431, 95% CI: 1.245-9.455) and abscess with a larger diameter ( OR=1.664, 95% CI: 1.258-2.220) were associated with a higher risk of developing sepsis (all P<0.05). Mediation model showed that neutrophil count and triglyceride were the mediating variables of sepsis in patients with PLA. Conclusion:Comorbidity of diabetes is an independent risk factor of developing sepsis in patients with pyogenic liver abscess. Diabetes may induce sepsis by affecting the neutrophils and triglyceride.

Objective:To study the logistic regression model and Chi-square automatic interaction detection decision tree model in the prediction of the recurrence of acute pancreatitis (AP).Methods:Clinical data of 364 patients with AP admitted to the Affiliated Hospital of Jiangnan University from June 2021 to June 2022 were retrospectively analyzed, including 219 males and 145 females, aged 53 (19-91) years. The patients were divided into the recurrence group ( n=63), those who experienced a second or more episodes of AP, and the initial group ( n=301), those who were diagnosed of AP for the first time. Univariate and multivariate logistic regression analyses were performed to identify the factors associated with recurrence of AP, and the decision tree model was used to analyze those factors. Receiver operating characteristic (ROC) curve were plotted to analyze the predictive performance of the two models. Results:Multivariate logistic regression analysis showed that age ( OR=0.969, 95% CI: 0.949-0.990, P=0.004), body mass index ( OR=1.142, 95% CI: 1.059-1.232, P=0.001), and hyperlipidemia ( OR=3.034, 95% CI: 1.543-5.964, P=0.001) were independent factors influencing the recurrence of AP. The accuracy of the model in predicting recurrence was 83.2% (303/364). The decision tree model showed that hyperlipidemia and body mass index were factors influencing the recurrence of AP, with an accuracy of 82.7% (301/364) in predicting recurrence. The area under the ROC curve was larger in the logistic regression model compared to that in the decision tree model (0.776 vs 0.730, Z=2.02, P=0.043). Conclusion:The logistic regression model and the Chi-square automatic interaction detection decision tree model can help predict the recurrence of AP. It is recommended to combine the two models to better guide clinical practice.

Objective:To explore the prognosis and treatment experience of fetal/neonatal ovarian cyst.Methods:Clinical data of 35 cases of fetal/neonatal ovarian cyst (38 ovarian cysts) admitted to Guangdong Women and Children Hospital from June 2014 to December 2019 were retrospectively collected, including the cyst size before and after birth, ultrasonic features, intraoperative conditions, and pathology. According to the ultrasonic features at the first prenatal detection, the ovarian cysts were divided into two groups: simple cyst group (25 cysts) and complex cyst group (13 cysts). Two independent samples t-test and Fisher exact test were used to compare the characteristics of cysts between the two groups. The outcomes and treatment experience were summarized. Results:(1) The ratio of intraoperative torsion in the complex cysts group was higher than that in the simple cysts group [10/13 vs 32% (8/25), Fisher exact test, P<0.05]. (2) Twenty-five simple cysts were found on the first prenatal ultrasound scan, and 32% (8/25) of them eventually transformed into complex cysts. Among these eight cysts, the maximum diameter of five cysts was >4 cm before the transformation. (3) Postnatal ultrasound found one cyst regressed spontaneously and among the remaining 37 cysts, simple and complex type cysts were accounted for 16 and 21, respectively. Among the complex type cysts, 90% (19/21) were consistent with prenatal ultrasound. (4) Out of the 21 complicated cysts, 19 were surgically removed; the remaining two cysts (maximum diameter <3 cm) were observed conservatively and disappeared spontaneously within one year. During the operation, 81% (17/21) of the complicated cysts were found with torsion and 24% (5/21) with ovarian loss. Conclusions:Simple cysts can transform into complex cysts, especially the biggest diameter >4 cm. Complex fetal/neonatal ovarian cysts indicated by ultrasonography were more prone to torsion, which required postnatal operation.

Objective:To summarize the perinatal management and prognosis of congenital chylous ascites (CCA).Methods:Clinical data of 20 infants diagnosed with CCA and treated in Guangdong Women and Children Hospital from June 2015 to November 2020 were retrospectively analyzed and described.Results:There were ten patients with isolated CCA and ten with non-isolated CCA. In isolated CCA cases, seven were cured after conservative treatment and three after postoperative conservative treatment following an ineffective surgery. Non-isolated CCA cases were complicated by intrauterine cytomegalovirus infection ( n=1), malrotation of intestine ( n=4) or bilateral chylothorax ( n=5). In addition to conservative treatment for CCA, non-isolated CCA patients also received antiviral therapy, Ladd's procedure or intrauterine/extrauterine closed thoracic drainage. Of eight patients who were firstly diagnosed with ascites before 30 gestational weeks, including four isolated and four non-isolated cases, only one underwent surgical intervention. During hospitalization, serious infections occurred in three infants with isolated CCA and two with non-isolated CCA, and were all controlled by anti-infection treatment. During a follow-up to median age of 29 months (15-82 months), none of the patients had any abnormalities except for the one with intrauterine cytomegalovirus infection who was deaf at the age of two. Conclusions:Conservative management is effective and the prognosis is generally good for most cases with isolated CCA. Treatment and prognosis of non-isolated CCA depend on its comorbidities. Gestational age at diagnosis may not be a prognostic predictor.

BackgroundThe diagnosis of Alzheimer's disease （AD） still faces great challenges， and the advantage of electroencephalogram （EEG） diagnosis lies in its portable and non-invasive nature， so the EEG diagnosis of AD has occupied an important place in clinical research. ObjectiveTo evaluate the value of resting state EEG for AD diagnosis， and to provide references for early recognition of AD in clinical practice. MethodsClinical data of AD patients （n=59） in an Inpatient Geriatric Psychiatry Unit of Shenzhen Kangning Hospital from May 2019 to May 2022 were retrospectively analyzed， and healthy elderly individuals attending outpatient clinics at the hospital during the same period were enrolled as control group （n=54）. Eight-channel resting state EEG data were acquired， and the absolute power values in the α， β， θ and δ frequency bands and the α/θ ratio were obtained and calculated using Fast Fourier Transform （FFT）. Cognitive function assessments of patients were done by Mini-Mental State Examination （MMSE） and Montreal Cognitive Assessment （MoCA）. Spearman correlation analysis was used to examine the correlation between EEG findings and MMSE and MoCA scores of AD patienrs. Logistic regression prediction model for AD was built using currently available EEG and clinical variables， and the model performance was assessed using the receiver operating characteristic （ROC） curve and the area under curve （AUC）. ResultsThe θ-band absolute powers in the right mid-frontal （F4） and mid-lateral （F7， F8） regions were higher in AD patients than those in healthy controls， with statistically significant difference （t=-2.844， -2.825， -3.014， P<0.05 or 0.01）. The absolute powers of α/θ ratio in prefrontal （Fp1， Fp2）， mid-frontal （F3， F4） and mid-lateral （F7， F8） regions showed a notable reduction in AD patients compared with healthy controls， with statistical difference （t=2.081， 2.327， 3.423， 2.358， 3.272， 2.445， P<0.05 or 0.01）. Spearman correlation analysis denoted that MMSE score was positively correlated with the absolute powers of α-band， β-band and α/θ ratio （r=0.206， 0.288， 0.372， P<0.05 or 0.01）. MoCA score was positively correlated with β absolute powers and α/θ ratio （r=0.201， 0.315， P<0.05 or 0.01）， and negatively correlated with θ absolute power （r=-0.218， P<0.05）. ROC curve revealed an AUC of 0.882 （95% CI： 0.820~0.943）， a sensitivity of 0.966 and a specificity of 0.673 for the AD prediction model based on EEG variables， while the prediction model for AD using comprehensive variables achieved better predictive efficacy， reaching an AUC， sensitivity and specificity of 0.946 （95% CI： 0.905~0.986）， 0.948 and 0.873， respectively. ConclusionResting state EEG of AD patients is correlated with cognitive function， and are of great value in the diagnosis of AD， with θ absolute power and α/θ ratio in EEG being the most strongly correlated with AD.