Objective:To analyze the real clinical effects and prognosis of neoadjuvant therapy for locally advanced resectable esophageal cancer.Methods:Two hundred and one patients with locally advanced resectable esophageal cancer who underwent different neoadjuvant treatments at Nanchong Central Hospital of Sichuan Province from January 2019 to December 2021 were retrospective analyzed. Patients were divided into neoadjuvant chemoradiotherapy group ( n=87), neoadjuvant immunochemotherapy group ( n=69), and neoadjuvant chemotherapy group ( n=45) according to the different methods of neoadjuvant therapy. Patients underwent surgical treatment 4-6 weeks after completing neoadjuvant therapy. The postoperative pathological response of three groups of patients were compared. Kaplan-Meier method was used to draw survival curves. Log-rank tests were performed to analyze overall survival (OS) rate, local recurrence-free survival (LRFS) rate, and distant metastasis-free survival (DMFS) rate in three groups of patients. Results:The pathological complete response rates of the neoadjuvant chemoradiotherapy group, neoadjuvant immunochemotherapy group and neoadjuvant chemotherapy group were 33.3% (29/87), 40.6% (28/69) and 13.3% (6/45), respectively, with a statistically significant difference ( χ2=9.68, P=0.008). The pathological complete response rates in the neoadjuvant chemoradiotherapy group and neoadjuvant immunochemotherapy group were higher than that in the neoadjuvant chemotherapy group, with statistically significant differences ( χ2=6.09, P=0.014; χ2=9.66, P=0.002) ; there was no statistically significant difference between the neoadjuvant chemoradiotherapy group and the neoadjuvant immunochemotherapy group ( χ2=0.87, P=0.351). The major pathologic response rates of the three groups were 58.6% (51/87), 59.4% (41/69) and 31.1% (14/45), respectively, with a statistically significant difference ( χ2=10.89, P=0.004). The major pathologic response rates of the neoadjuvant chemoradiotherapy group and neoadjuvant immunochemotherapy group were significantly higher than that of the neoadjuvant chemotherapy group, with statistically significant differences ( χ2=8.98, P=0.003; χ2=8.74, P=0.003) ; there was no statistically significant difference between the neoadjuvant chemoradiotherapy group and the neoadjuvant immunochemotherapy group ( χ2=0.10, P=0.920). The 3-year OS rates of the three groups were 43.7%, 42.0% and 33.3%, respectively, with no statistically significant difference ( χ2=0.79, P=0.347). The 3-year LRFS rates of the three groups were 67.8%, 66.7% and 46.7%, respectively, with a statistically significant difference ( χ2=7.58, P=0.023), the LRFS rates in the neoadjuvant chemoradiotherapy group and neoadjuvant immunochemotherapy group were significantly higher than that in the neoadjuvant chemotherapy group, with statistically significant differences ( χ2=4.17, P=0.041; χ2=4.15, P=0.042) ; there was no statistically significant difference in LRFS rates between the neoadjuvant chemoradiotherapy group and the neoadjuvant immunochemotherapy group ( χ2=0.01, P=0.923). The 3-year DMFS rates of the three groups were 37.9%, 37.7% and 28.9%, respectively, with no statistically significant difference ( χ2=0.14, P=0.707) . Conclusion:Different neoadjuvant therapies have different therapeutic effects in the treatment of locally advanced resectable esophageal cancer, neoadjuvant chemoradiotherapy and neoadjuvant immunochemotherapy can achieve a better pathological response rates and LRFS rates.

Objective To observe the correlation between growth impairment induced by long-term oral glucocorticoids(GC)therapy and the ratio of FGF23/Klotho in children with primary nephrotic syndrome(PNS).Methods A prospective study was conducted on 56 children with GC-sensitive PNS who had discontinued GC therapy for more than 3 months and revisited the Department of Pediatrics of the First Affiliated Hospital of Henan University of Traditional Chinese Medicine between June 2022 and December 2022.After monitoring qualitative and quantitative urine protein levels upon admission,the children with proteinuria relapse were treated with GC(GC group;n=29),while those without relapse did not receive GC treatment(non-GC group;n=27).In addition,29 healthy children aged 3 to prepuberty were selected as the control group.Height,bone age,growth rate,and the FGF23/Klotho ratio were compared among the groups.The correlations of the FGF23/Klotho ratio with height,bone age,and growth rate were analyzed.Results The FGF23/Klotho ratio in the GC group was significantly higher than that in the non-GC group after 1 month of GC therapy(P＜0.05),and the height and bone age growth rates within 6 months were lower than those in the non-GC group(P＜0.05).Correlation analysis showed significant negative correlations between the FGF23/Klotho ratio after 1 month of treatment and the growth rates of height and bone age within 6 months in children with PNS(r=-0.356 and-0.436,respectively;P＜0.05).Conclusions The disturbance in FGF23/Klotho homeostasis is one of the mechanisms underlying the growth impairment caused by long-term oral GC therapy.[Chinese Journal of Contemporary Pediatrics,2024,26(3):269-2741

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.

To carry out phylogenetic analysis for drug-resistance genes from clinical isolates of Helicobacter pylori (Hp) among patients with gastric diseases from Tibet, China.

METHODSHp strains were isolated and cultured from saliva and gastric mucosal tissues derived from patients with gastric diseases. Nine strains (including 5 isolated from oral tissues, 1 isolated from gastric tissues, and 3 representative strains of SS international standard strains used for animal models) were tested for common antibiotic resistance. Together with an ACTT 11637 international standard strain, these were subjected to re-sequencing to obtain drug-resistance genes. Such genes from various sources were compared with the resistance genes of Hp strains recorded by the NCBI website. Combined with results of drug-resistance experiments, correlation between molecular evolution and drug-resistance was analyzed.

RESULTSTesting of gastric mucosal tissues and salivary samples from 217 patients has found 89 Hp strains, which yielded a total infection rate of 41.01%. The resistance rates of 9 representative Hp strains for clarithromycin, amoxicillin, metronidazole, levofloxacin and tetracycline were 77.8%, 77.8%, 44.4%, 77.8%, and 77.8%, respectively. Compared with the reference strain, the similarity between clarithromycin-resistance genes was 99%, and that between amoxicillin- and metronidazole-resistance genes was 96%-97%. A2143G mutation was also found in clarithromycin-resistant genes of three Hp strains.

CONCLUSIONThe sensitivity of Hp to metronidazole is much higher in patients from Tibet region, and the sensitivity of Hp to clarithromycin, amoxicillin, levofloxacin and tetracycline is poor. Resistance mutations are consistent with drug resistance.

AIM:To investigate the effect of RNA interference (RNAi)-mediated insulin-like growth factor 1 receptor ( IGF1R) gene silencing on the growth , migration, and invasion of hepatocellular carcinoma cells .METHODS:The most effective siRNA targeting IGF1R gene was designed and screened .After lentiviral expression vector pLVX-shR-NA2-IGF1R carrying the most effective siRNA sequence was constructed , it was transfected into 293T cells and packed into pLVX-shRNA2-IGF1R lentivirus.Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R lentivirus to screen the positive clone Huh7 cells and Hep3B cells with the lentivirus .These Huh7 cells and Hep3B cells were cultured to ana-lyze the mRNA level of IGF1R, cell proliferation, cell cycle, cell apoptosis, cell migration/invasion, and the protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL.RESULTS:The mRNA expression of IGF1R in Huh7 cells and Hep3B cells with pLVX-shRNA2-IGF1R lentivirus was significantly reduced .The proliferation of these cells was remarkably inhibited , and the number in G 1 phase was increased significantly .The percentages of apop-totic cells were increased markedly , and the number of cell migration/invasion was decreased markedly .The protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL were decreased significantly compared with the blank control group and negative control group .CONCLUSION:The RNAi-mediated IGF1R gene silencing sig-nificantly suppresses the growth and the malignant biological characteristics of Huh 7 cells and Hep3B cells, which may be involved in the reduced protein levels of the above genes induced by down -regulation of IGF1R expression.

Periampullary carcinoma is a rare malignant tumor of digestive system,and its accurate diagnosis is still difficult.From January 2007 to July 2012,12 patients with periampullary carcinoma had been admitted to the Southwest Hospital of Third Military Medical University,and the imaging data were retrospectively analyzed.The results of ultrasonography revealed that all tumors were hypoechoic.The tumor displayed hyperenhancement in 3 patients,isoenhancement in 1 patient,hypoenhancement in 8 patients during the arterial phase on contrastenhanced ultrasonography (CEUS),while the tumor displayed hypoenhancement in all patients during the venous phase.Magnetic resonance imaging (MRI) plain scanning showed duodenal papilla enlargement in 1 patient,ampullary tissue mass signal in 2 patients,tissue mass signal at distal common bile duct in 2 patients,the rest 7 patients did not show tissue mass signal.Lower biliary obstruction was the common manifestation of the 12 patients on magnetic resonance cholangiopancreatography (MRCP),intrahepatic and extrahepatic bile vine-like expansion in 4 patients,double duct sign in 2 patients,the bottom of common bile duct with filling defect in 2 patients and it revealed beak-like narrow in 1 patient.CEUS,MRI and MRCP could both play an important role as conventional methods in diagnosing periampullary carcinoma.

Ex-vivo liver resection is developed based on liver transplantation and technique of cold preservation of organs.It overcomes the shortcomings of time limit of warm ischemia and high technique demand of hepatectomy of tumors located at critical sites.A 58-year-old woman with hepatocellular carcinoma located close to the middle hepatic vein combined with invasion of right hepatic vein was admitted to Southwest Hospital.Because of the critical tumor site,conventional liver resection Wag assessed as impossible.Ex-vivo liver resection was performed,and a vessel patch from an organ wag harvested to repair the defect of the right hepatic vein,and then the liver remnant was subsequently autotransplanted.After operation,the patient recovered smoothly without venous outflow complication.Bile leakage wag observed on postoperative day 23,and the maximnm volume of intraperitoneal drainage wag 200 ml per 24 hours.Endoscopic nasobiliary drainage Was performed and the volume of intraperitoneal drainage gradually decreased to none.Liver function of the patient was back to normal and with no tumor recurrence at the end of 6 months of follow up.Ex-vivoliver resection is beneficial to patients with centrally located hepatocellular carcinoma with the involvement of hepatic vein and inferior vena cava.

With the genomic DNA of strain EJS-3 as the template, we amplified the gene of fibrinolytic enzyme from Paenibacillus polymyxa (PPFE-I) by PCR. We purified the PCR product and ligated it into pMD19-T. After DNA sequencing, we cloned the PPFE-I gene into expression vector pET-DsbA and transformed it into Escherichia coli BL21(DE3). Upon induction of IPTG, we found that the activity of recombinant fibrinolytic enzyme fused with DsbA expressed in Escherichia coli was 228 IU/mL. SDS-PAGE analysis showed that the recombinant enzyme was soluble and accounted for about 18.4% of total cell protein. Western blotting demonstrated that the recombinant protein was DsbA-PPFE-I. We purified the recombinant enzyme by Ni affinity chromatography, thrombin digestion and sephadex G-100 gel-filtration, and identified the molecular weight of purified product to be 66.3 kDa with MALDI-TOF mass spectrometry. The purified enzyme exhibited distinct fibrinolytic activity on fibrin plate.
Antifibrinolytic Agents ; pharmacology ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fibrinolytic Agents ; metabolism ; Genetic Vectors ; genetics ; Paenibacillus ; chemistry ; enzymology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

Cold perfusion of liver can significantly alleviate the ischemia-reperfusion injury caused by hepatic blood flow occlusion. We have modified the technique of cold perfusion of liver and applied it to total pancreatectomy for patients with pancreatic head carcinoma complicated with metastasis to the body and tail of pancreas and with portal invasion. After skeletonization of the hepatoduodenal ligament, the amputation of the portal vein and blockage of the superior mesentoric vein were performed before portal perfusion. Meanwhile, pancreatic head resection, duodenectomy, subtotal gastrectomy and partial resection of the superior mesenteric vein and portal vein were carried out. Superior mesenteric vein and portal vein bypass grafting was achieved with artificial vessels. The digestive tract was reconstructed after it was freed of the spleen and resection of the body and tail of pancreas to the left side of superior mesenteric vein, greater omentum and intestine from the end of the colon to splenic flexure of colon. The patient was followed up for 3 months, and the general condition was good, although diarrhea frequently occurred. No tumor metastasis occurred.

Lipases are important biocatalysts that are widely used in food processing and bio-diesel production. However, organic solvents could inactivate some lipases during applications. Therefore, the efficient cloning and expression of the organic solvent-tolerant lipase is important to its application. In this work, we first found out an organic solvent-tolerant lipase from Staphylococcus saprophyticus M36 and amplified the 741 bp Lipase gene lip3 (GenBank Accession No. FJ979867), by PCR, which encoded a 31.6 kD polypeptide of 247 amino acid residues. But the lipase shared 83% identity with tentative lip3 gene of Staphylococcus saprophyticus (GenBank Accession No. AP008934). We connected the gene with expression vector pET-DsbA, transformed it into Escherichia coli BL21 (DE3), and obtained the recombinant pET-DsbA-lip3. With the induction by 0.4 mmol/L of isopropyl beta-D-thiogalactopyranoside at pH 8.0, OD600 1.0, 25 degrees C for 12 h, the lipase activity reached up to 25.8 U/mL. The lipase expressed was stable in the presence of methanol, n-hexane, and isooctane, n-heptane.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Lipase ; biosynthesis ; genetics ; Molecular Sequence Data ; Organic Chemicals ; chemistry ; Recombinant Proteins ; biosynthesis ; genetics ; Solvents ; chemistry ; Staphylococcus saprophyticus ; enzymology