We produced α-ketoglutaric acid (α-KG) from L-glutamic acid, using enzymatic transformation approach with L-glutamate oxidase (LGOX). First, wild strain Streptomyces sp. FMME066 was mutated with NTG, a genetically stable mutant Streptomyces sp. FMME067 was obtained. Under the optimal nutrition conditions with fructose 10 g/L, peptone 7.5 g/L, KH2PO4 1 g/L and CaCl2 0.05 g/L, the maximum LGOX activity reached 0.14 U/mL. The LGOX was stable to pH and temperature, and Mn2+ had a stimulating effect. Finally, after 24 h enzymatic conversion under the optimal conditions, the maximum titer of α-KG reached 38.1 g/L from 47 g/L L-glutamic acid. Enzymatic transformation by LGOX is a potential approach for α-KG production.
Amino Acid Oxidoreductases ; metabolism ; Fermentation ; Glutamic Acid ; metabolism ; Ketoglutaric Acids ; metabolism ; Streptomyces ; genetics ; metabolism

Objective:To compare the accuracy and timeliness of 3-lead electrocardiography (ECG) and pulse oximetry (POX) in neonatal heart rate (HR) monitoring after birth.Methods:This prospective study recruited 42 high-risk newborns with gestational age ≥37 weeks and birth weight >1 500 g who were born through cesarean section without resuscitation requirement in Xi'an People's Hospital (Xi'an Fourth Hospital) from October 2019 to August 2020. 3-lead ECG electrodes and POX sensors were attached to the neonates immediately after drying to continuously monitor the HR within 10 min after birth. All procedure was recorded by video camera, and data were independently analyzed by a clinician after the procedure was completed. Differences in time required to connect the devices, time to obtain a reliable HR and the interval between them, the time needed for obtaining a reliable HR after birth, the proportion of neonates with reliable HR obtained within 5 min after birth and the consistency in the reliable HR readings between the two devices were compared using Wilcoxon signed-rank test, McNemar test, Spearman's correlation coefficient, intraclass correlation coefficient or Bland-Altman bias analysis.Results:The median time required to connect POX and 3-lead ECG and to acquire a reliable HR were 13.0 s (10.0-17.0 s) vs 23.0 s (18.0-28.3 s) ( Z=-5.050, P<0.001), and 79.5 s (56.2-128.0 s) vs 11.0 s (10.0-13.3 s) ( Z=-5.646, P<0.001), respectively. The total time from the beginning of connecting the devices and birth to acquiring a reliable HR were both longer for POX than those for 3-lead ECG [92.0 s (71.3-139.0 s) vs 35.0 s (30.0-39.5 s), Z=-5.579, P<0.001; 110.5 s (85.8-153.5 s) vs 52.0 s (45.0-66.3 s), Z=-5.579, P<0.001]. Reliable HRs were obtained in 69.1% (29/42) and 2.4% (1/42) of the infants by 3-lead ECG and POX within 1 min after birth, respectively. The percentage of infants for obtaining a reliable HR detected by 3-lead ECG within 5 min after birth were more than those by POX, but with statistically significant differences only at the first 60 s, 90 s, 120 s and 150 s (all P<0.001). The median HRs obtained by 3-lead ECG and POX within 10 min after birth were 161 beats/min (147-175 beats/min) and 160 beats/min (146-176 beats/min), respectively ( r=0.966, P<0.001). The mean difference of HR detected by the two devices was 0.56 beats/min (95% CI:-4.3 to 5.4 beats/min). The intraclass correlation coefficient was 0.961, showing good internal consistency. Conclusions:Neonatal HR can be assessed accurately by 3-lead ECG within 1 min after birth, which is far earlier than that by POX. Therefore, 3-lead ECG can be an option for continuously HR monitor in neonatal resuscitation.

Objective To assess the perioperative safety and postoperative function of the pure 3-dimensional laparoscopic cystectomy and urinary diversion (P3DLC-UD).Methods From April 2014 to July 2015,P3DLC-UD was performed in 15 patients diagnosed with the bladder cancer in our center (orthotopic ileal neobladder for 8 cases and ileal conduit for 7 cases).Perioperative data,postoperative continence and overall survival rate were retrospectively analyzed.Results Fifteen patients underwent P3DLC-UD successfully as planned,with 8 patients underwent orthotopic ileal neobladder and 7 patients underwent ileal conduit.In neobladder group,the operative time were 300-600mmin (mean 428 min),estimated blood loss were l 00-400ml (mean 210mml),and dissected lymph nodes were 11-29 (mean 16).One patient required blood transfusion (800ml) and one patient was diagnosed of constipation.The patients were followed up for a median period of 10 months (3-15 months).The renal function was normal with serum creatinine of 36.4-99.0 μ mol/L (mean 77.3 μmol/L).One patient died of intestinal obstruction and 8 patients had no recurrence in neobladder group.As to postoperative continence,only 2 patients demanded 1 pad at daytime,while all patients needed 1 pad at nighttime.In ileal conduit group,the operative time were 300-390 min (mean 354 min),estimated blood loss were 50-400ml (mean 190ml),and dissected lymph nodes were 9-41 (mean 22),while 9-41 (mean 19) lymph nodes were got for all 15 cases.Two patients were diagnosed with urinary infection after the surgery.The patients were followed up for a median period of 5 months (1-9 months).The renal function was normal with serum creatinine of 36.4-74.0 μmol/L (mean 60.8 μmol/L).One patient died of cerebral infarction,and 1 patient found distant metastases in lung and died of cancer after chemotherapy during the follow-up period.There were no recurrent tumors of the other 5 patients in ileal conduit group.Conclusions P3DLC-UD is safe and feasible.More extensive,longer-term randomized trials are required to comprehensively assess the appropriateness and potential of this technique.

Objective:To analyze the learning effect of laparoscopic radical cystectomy(LRC)+ modified ileal conduit(MIC).Methods:From 2014 to 2019, 42 patients underwent MIC and their clinical data was retrospectively analyzed. 34 operations were performed by surgeon 1 and 8 operations by surgeon 2. We divided the 34 patients of surgeon 1 into three groups according to their surgical sequence (group A, 1st to 12th; group B, 13th to 23th; group C, 24 th to 34 th), the 8 cases of surgeon 2 was regarded as group D. The history of abdomen surgery in the 4 groups were 0, 1, 4, 3 cases, respectively ( P<0.05). There was no significant difference of the other baseline characteristics, such as age, BMI, American Society of Anesthesiologists. Then we compared several variables between the 4 groups like operation time, time of ileal conduit construction, blood loss, complication rate, lymph node yield, surgical margin, etc. The key steps of the MIC included isolating terminal ileum when the mesentery was transilluminated, performing end-to-end reflux ureterointestinal anastomosis after the efferent loop was fixed, closing the rent of the retroperitoneum. Results:All operations were performed intracorporeally with no transition to open surgery. The operative time for group A, B, C were 330.0(320.0, 360.0)min, 300.0(250.0, 308.0)min, 270.0(216.0, 324.0)min, respectively ( P =0.010). The time of ileal conduit construction of the 3 groups were 136.5(131.3, 147.5)min, 92.0(79.0, 119.0)min, 79.0(72.0, 115.0)min, respectively ( P <0.001). In addition, the difference of the two variables above between A and B, A and C groups separately reached statistical significance ( P<0.05), while the difference between B and C groups did not ( P>0.05). Other variables, such as blood loss, complication rate, lymph node yield, surgical margin, between the 3 groups reached no statistical significance ( P>0.05). The operative time of group D was 420.0(350.0, 450.0)min, and it reached statistical significance ( P<0.05) when compared with group A. There were no significant differences in other variables, such as blood loss, complication rate, lymph node yield, surgical margin, among the 2 groups ( P>0.05). Conclusions:The learning effect of LRC+ MIC was obvious. When surgeon volume increased, the operative time decreased significantly. Variables like estimated blood loss and complication rate of the 2 surgeons did not reached significant difference, which indicated reproductivity and safety of this procedure.

Abstract: Objective To investigate the prevalence and influencing factors of depressive symptoms among primary and middle school students in Xihu District, Hangzhou City, so as to provide insights into depression control among primary and middle school students. Methods: Students were sampled from two primary schools, two junior high schools, two ordinary high schools and one vocational high school in Xihu District using a stratified cluster random sampling method from September to November, 2021. Participants' demographics, diet, exercise, sleep and campus bullying were collected using questionnaire surveys. The depressive symptoms were evaluated using the Chinese version of the Center for Epidemiologic Studies-Depression Scale (CES-D), and factors affecting the depressive symptoms were identified among primary and middle school students using a multivariable logistic regression model. Results: A total of 1 518 students were investigated, including 518 primary school students (34.12%), 363 junior high school students (23.91%), 463 ordinary high school students (30.50%) and 174 vocational high school students (11.46%). Of all participants, there were 805 males (53.03%). The prevalence of depressive symptoms was 38.87% among the participants, and was 36.10%, 42.42%, 42.33% and 30.46% among primary school, junior high school, ordinary high school and vocational high school students, respectively. The multivariable logistic regression analysis showed that gender (OR=1.601, 95%CI: 1.289-1.989), type of school (junior high school, OR=1.299, 95%CI: 0.974-1.734; ordinary high school, OR=1.351, 95%CI: 1.025-1.780; vocational high school, OR=0.862, 95%CI: 0.585-1.272), frequency of fresh vegetable intake (less than once daily, OR=0.454, 95%CI: 0.304-0.679; once and more daily, OR=0.359, 95%CI: 0.235-0.548), duration spent on the internet (0.5 to 1 h daily, OR=1.447, 95%CI: 1.044-2.006; 1 h and longer daily, OR=1.456, 95%CI: 1.072-1.978), sufficient sleep (OR=0.525, 95%CI: 0.371-0.744), suffering from campus bullying (OR=1.992, 95%CI: 1.491-2.661) were associated with the development of depressive symptoms. Conclusions The prevalence of depressive symptoms is high among primary and middle school students in Xihu District. Gender, type of school, dietary behaviors, sleep duration and campus bullying are factors affecting the development of depressive symptoms.

Objective To explore the diagnosis rate.pathology types and positive rate of cancer cell in spu-tum of early central pulmonary carcinoma in three obstructive signs on chest X ray screened by fiberbromchoscope.Methods 326 cases of three obstructive signs with high risk of lung cancer were screened for central pulmonarycarcinoma by spiral CT.biopsy by fiberbronchoscope and cytological examination of sputum.Results 32 patients were diagnosed with central pulmonary carcinoma,with morbidity of 9.8%.In these patients,21 were confirmed with obstructive pneumonia(65.6%),7 with obstructive atelectasis(21.9%),4 with obstructive emphysema(12.5%);In terms of pathology type,16 cases were defined as squamous cell carcinoma(50.0%),9 as small cell carcinoma(28.1%).3 were as large cell carcinoma(9.4%).2 were as adenocarcinoma(6.3%),1 as admosquamous carci-noma(3.1%),1 as bronchial gland carcinoma(3.1%);cancer cell could be found in sputum of 5 patients of 32 cases,among them,it was found in 3 of 21 patients with obstructive pneumonia(14.3%),1 in 7 patients with ob-structive atelectasis(14.3%),1 in 4 patients with obstructive emphysema(25.0%).Conclusion The prevelance of early central pulmonary carcinoma in three obstructive signs on chest X-ray is 9.815%,in which squamous carci-noma and small-cell carcinoma are common in pathology type.Screening can increase the detection rate of early pul-monary carcinoma.

Objective To explore the association between serum uric acid (SUA) and all-cause mortality in men. Methods In this prospective cohort study,data being used was derived from the Kailuan study cohort. A total of 81 110 male workers who had taken part in the Kailuan physical examination were enrolled. Subjects with previous myocardial infarction,stroke,cancer, eGFR<30 ml/(min·1.73 m2)accidental deaths and those ever used drugs that seemed to have showed an effect on blood uric acid,were excluded. All the information was gathered from a unified questionnaire,measured by blood biochemistry and with the mean period of follow up as(47.5±4.3) months. Based on the 2006-2007 SUA value,observed objects were divided into five groups,with multivariate Cox proportional hazard regression analysis used to estimate the relationship between SUA and all-cause mortality in men. Results 1)At the end of the follow-up period in 2010-2011, the number of deaths were 315,278,243,292 and 341 among the different SUA quinte,with incidence rates of all-cause mortality as 2.43%,2.36%,1.96%,2.42%and 2.92%,respectively. 2)Data from the Single factor Cox proportional hazard regression analysis showed that,when comparing with the third quinte,HR values of the all-cause mortality were 1.32(1.11-1.56),1.19(1.00-1.41),1.20(1.01-1.43)and 1.41(1.19-1.66)in other four groups,respectively. 3)When factors were adjusted for age, systolic blood pressure,diastolic blood pressure,body mass index,triglyceride,total cholesterol, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, fasting glucose, high-sensitivity C-reactive protein,smoking history and history of drinking,education,profession, economy,etc.,results from the Multiple Cox proportional hazard regression analysis showed the HR values of the all-cause mortality were 1.26(1.06-1.51),1.20(1.01-1.44),1.25(1.05-1.49),1.42 (1.19-1.68) in other four groups,respectively,comparing to the third quinte. Conclusion Using SUA as the independent risk factor of all-cause mortality,the exceptional levels of SUA were associated with an increasing risk for all-cause mortality while the association of SUA with all-cause mortality appeared an“U”shaped curve.

Objective: To explore the role of toll-like receptor 4 (TLR4)/NF-κB signaling pathway in acute necrotizing pancreatitis (ANP)-associated lung injury and the intervention of lipoxin A4 (LXA4) analogue. Methods: Forty-five Sprague-Dawley rats were randomly(random number)divided into the sham operation group, experimental group, and intervention group, each group containing 15 rats. ANP animal models were prepared by injecting sodium taurocholate into biliopancreatic tube in the experimental group. No sodium taurocholate was injected into biliopancreatic duct in the sham operation group. After the preparation of ANP animal models in the intervention group, LXA4 was injected through the tail vein. Rats in each group were randomly divided into 3 subgroups (n=5 each subgroup). The serum amylase, TNF- α, IL-1β, IL-6 and endotoxin levels were detected 6, 12 and 24 h after the operation. The lung injury scores were assessed and the lung wet/dry weight ratio was calculated. The expressions of TLR4 and NF-κB p65 in lung tissues were detected by Western blot. Results: Serum levels of amylase, TNF-α, IL-1β, IL-6 and endotoxin in the experimental and intervention groups were significantly higher than those in the sham operation group, while the levels of the above indicators in the intervention group was significantly lower than those in the experimental group, and the differences were statistically significant (P<0.05). Postoperative lung injury scores and lung wet/dry weight ratio in the experimental group were significantly higher than those in the sham operation group, and the differences were statistically significant (P<0.05). Lung injury scores in the intervention group 6 h after operation had no significant difference compared with those in the sham operation group (P>0.05), while lung wet/dry weight ratio in the intervention group 6 h after operation, and lung injury scores and lung wet/dry weight ratio in the intervention group 12 h or 24 h respectively after operation were significantly higher than those in the sham operation group, with statistically significant differences (P<0.05). Postoperative lung injury scores and lung wet/dry weight ratio in the intervention group were significantly lower than those in the experimental group, and the differences were statistically significant (P<0.05). The expressions of TLR4 and p65 in the lung tissues of the experimental and intervention groups were significantly higher than those of the sham operation group, and the expressions of TLR4 and p65 in the lung tissues of the intervention group were significantly lower than those of the experimental group, with statistically significant differences (P<0.05). Conclusions LXA4 can reduce the severity of acute necrotizing pancreatitis-associated lung injury, and its mechanism is related to reducing the stimulation of endotoxin, thus inhibiting TLR4 signaling pathway and the activation of p65 to down-regulate the level of pro-inflammatory cytokines.

Objective:To explore the mechanism of pioglitazone in reducing lung injury induced by acute pancreatitis.Methods:Thirty healthy male SD rats were randomly(random number) divided into the sham operation group, model group and pioglitazone group, with 10 rats in each group. After anesthesia, the rats in the sham operation group were injected with normal saline retrogradely through the pancreaticobiliary duct. In the model group, after anesthesia, the rats were retrogradely injected with sodium taurocholate into the pancreaticobiliary duct to construct the lung injury model of severe acute pancreatitis. In the pioglitazone group, the model was established after intraperitoneal injection of pioglitazone. Six rats in each group were randomly selected and killed 12 h after operation, and then lung tissue and venous blood were collected. The levels of serum amylase and TNF-α and NO in lung tissue homogenate were detected and compared among the three groups; the expression of TLR2 mRNA and TLR4 mRNA in lung tissue was detected by RT-PCR and compared among the three groups; the lung tissue pathological injury score and lung leakage index were calculated and compared among the three groups. The correlation of TLR2 and TLR4’s mRNA expression with lung tissue pathological injury score and lung leakage index was analyzed.Results:The levels of serum amylase and the levels of TNF-α and NO in lung tissue homogenate in the model group were significantly higher than those in the sham operation group, and the above indexes in the pioglitazone group were significantly lower than those in the model group ( P<0.05). The expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue, the lung tissue pathological injury score and lung leakage index in the model group were significantly higher than those in the sham operation group, and the above indexes in the pioglitazone group were significantly lower than those in the model group ( P<0.05). Spearman correlation analysis showed that the expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue were significantly positively correlated with the lung tissue pathological injury score ( rs=0.959, P<0.001; rs=0.924, P<0.001). Pearson correlation analysis showed that the expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue were significantly positively correlated with the lung leakage index ( r=0.957, P<0.001; r=0.958, P<0.001). Conclusions:Pioglitazone may reduce the severity of severe acute pancreatitis induced lung injury by inhibiting the expression of TLR2 mRNA and TLR4 mRNA in lung tissue.

Objective:To explore the mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of pancreatic cells and participating in the occurrence and development of acute pancreatitis.Methods:Pancreatic acinar cells AR42J from rats were treated by caerulein, and then the levels of amylase, tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1) and IL-6 in the supernatant of cell culture were detected by ELISA to verify the cell model of acute pancreatitis. RT-PCR was used to detect the expression of miR-494 in normal AR42J cells (control group) and acute pancreatitis cell model (model group). Flow cytometry was used to detect the apoptosis of the control group, negative control miRNA transfected acute pancreatitis cell model (negative control group) and miR-494 transfected acute pancreatitis cell model (miR-494 transfection group). Western blot was used to detect the expression of ROCK1 and PTEN in the control group, negative control group and miR-494 transfection group.Results:The levels of amylase, TNF-α, IL-1 and IL-6 in the supernatant of AR42J cells treated with caerulein for 8 h and 12 h were significantly higher than those at 0 h and the control group ( P<0.05), indicating that the model was successfully constructed. The expression levels of miR-494 at 8 h, 12 h and 24 h after the establishment of acute pancreatitis cell model were significantly higher than those at 4 h and the control group ( P < 0.05). The apoptosis rate of the model group was significantly higher than that of the control group ( P<0.05), and the apoptosis rate of the miR-494 transfection group was significantly lower than that of the model group ( P<0.05). The expression levels of ROCK1 and PTEN in the miR-494 transfection group were significantly lower than those in the model group and negative control group ( P<0.05). Conclusions:When acute pancreatitis occurs, overexpression of miR-494 can inhibit the expression of pro-apoptotic protein, thus inhibiting the apoptosis of pancreatic acinar cells and promoting the development of acute pancreatitis.