Objective To analyze the epidemiological characteristics of intestinal flora in children with rotavirus enteritis (RVE) in Qinghai, and to provide a basis for clinical treatment and prevention. Methods A total of 64 children with RVE were enrolled in the RVE group, and 62 healthy children who received physical examination in our hospital during the same period were enrolled in the healthy control group. Stool specimens were collected, and fluorescence quantitative PCR technology was used to detect 16S rRNA of feces specimens for intestinal floras including Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus. The contents of common intestinal flora in two groups were compared. The changes in the contents of four major intestinal bacteria in RVE children of different genders, ages, nationalities and onset seasons were analyzed. Results Compared with the healthy control group, fecal contents of Bifidobacterium and Lactobacillus in the RVE group were significantly reduced (P<0.05). There was a significant difference in the fecal contents of Bifidobacterium and Lactobacillus in RVE children with different ages (P<0.05). The fecal contents of Escherichia coli and Enterococcus in children with RVE from 0 to 1 years old were significantly lower than those in other age groups (P <0.05); (P<0.05). Fecal Bifidobacterium content in RVE children of Han nationality was significantly higher than that of Tibetan children, while Lactobacillus content was significantly lower than that of Tibetan children (P<0.05). The fecal contents of Bifidobacterium and Lactobacillus of RVE children in autumn and winter were significantly lower than those in other seasons (P<0.05). Conclusion In Qinghai, rotavirus enteritis has a great influence on the contents of Bifidobacterium and Lactobacillus in children's intestinal tract. The contents of main intestinal bacteria are different in children with rotavirus enteritis of different ages, nationalities and seasons.

ObjectiveTo explore the mechanism of Tibetan medicine Ershiwuwei Guijiuwan against osteoporosis in ovariectomized rats through the classical Wnt/β-catenin signaling pathway. MethodForty-eight 3-month-old SD female rats were randomized into model group （equivalent volume of normal saline）， sham operation group （equivalent volume of normal saline）， estradiol group （0.1 mg·kg^-1 estradiol valerate）， and high-， medium-， low-dose Ershiwuwei Guijiuwan groups （800， 400， 200 mg·kg^-1， respectively）. For the modeling， some adipose tissue near the ovaries was removed in the sham operation group， and ovaries were excised in other groups. Administration （ig， once a day） started two weeks after the operation and lasted 12 weeks. After sampling， based on hematoxylin and eosin （HE） staining， the morphological changes of the right femur and lumbar spine of the rats were observed. The enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum levels of rat tartrate-resistant acid phosphatase 5b （TRAP5b）， collagen type Ⅰ C-terminal peptide （CTX-Ⅰ）， bone alkaline phosphatase （BALP）， amino-terminal propeptide of type Ⅰ procollagen （PINP）， and bone Gla-protein （BGP）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to examine the mRNA expression of β-catenin and Runt-related transcription factor 2 （Runx2） in femur， and Western blot to detect the protein expression of β-catenin， Runx2， and low-density lipoprotein receptor-related protein-5 （Lrp-5）. ResultCompared with the sham operation group， the model group showed disordered and sparse bone trabecula with fewer connections， decrease in serum levels of BALP， BGP， and PINP （P<0.01）， increase in levels of CTX-Ⅰ and TRAP5b （P<0.01）， reduction in mRNA expression of β-catenin and Runx2 in femoral tissue （P<0.01） and protein expression of Lrp-5， β-catenin， and Runx2 （P<0.01）. Compared with the model group， estradiol and each dose of Ershiwuwei Guijiuwan increased the volume of bone trabecula， made thebone trabecula closely arranged， reduced the loss of the trabecular meshwork， raised the serum levels of BALP and BGP （P<0.01）， and lowered TRAP5b level （P<0.01）. PINP level was significantly increased in the high-dose Ershiwuwei Guijiuwan group and estradiol group （P<0.01） and CTX-Ⅰ level was significantly decreased in the high-dose Erwenwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. The mRNA expression of β-catenin in femoral tissue and protein expression of Lrp-5 and β-catenin in estradiol group and three Ershiwuwei Guijiuwan groups were significantly increased （P<0.05， P<0.01）， and the levels of Runx2 mRNA and protein were significantly increased in the high-dose Ershiwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. ConclusionTibetan medicine Ershiwuwei Guijiuwan is effective for the osteoporosis in ovariectomized rats， which may be related to the classic Wnt/β-catenin signaling pathway. It affects bone metabolism by up-regulating the expression of osteogenesis-related genes in the signaling pathway.

Objective: To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice. Methods: Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group, and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group, and the same volume of normal saline solution was used in the model group and control group in the same way.The activities, mental state, body weight, and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention, the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin- eosin staining; serum homocysteine (Hcy) was detected by ELISA assay; the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR; the relative expressions of B lymphoma 2 protein (bcl-2), bcl-2 related X protein (bax), 3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay; superoxide dismutase (SOD), 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits, and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35). Results: Over the experimental period, The FPG was normal and body weight was gradually increased in the mice of control group.The FPG>16.7 mmol/L and the mice appeared obese.In the folic acid group, both body mass and FPG of the mice were gradually reduced.At the end of drug administration, serum Hcy concentration of the mice was (27.18±3.18)μmol/L in the model group, which was significantly higher than (8.28±2.18)μmol/L in the control group and (13.73±2.54)μmol/L in the folic acid group (all at P<0.05). The retinal structure was intact in the control group, and the retinas were thinning with more capillaries and inflammatory cells in the model group, the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group, and those in the folic acid group were reduced in comparison with the model group (all at P<0.05). The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group, and the relative expressions of bax, 3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P<0.05 ). The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group, and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P<0.05). The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group. Conclusions Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy, inhibiting oxidative stress and cell apoptosis.

Objective: To explore the drug resistance of Isolated From Blood Culture Escherichia coli (E. coli) in a hospital in Qinghai over the past seven years, to evaluate the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the homologous origin of E. coli, and to establish a protein fingerprint library to match with it, adjuvant clinical experience medication so as to provide the basis for the prevention and control of hospital-acquired infections. Methods: Retrospective analysis of blood cultures sent to hospitals from January 2016 to December 2022. Drug resistance and resistance changes in E. coli.A total of 1 841 E. coli strains were isolated from Qinghai Provincial People's Hospital from January 2016 to December 2022; all strains were identified by MALDI-TOF MS, and the VITEK2.0 drug sensitivity analyzer was applied for drug sensitivity analysis of the strains, and the mass spectrometry homology analysis and self-constructed protein fingerprint library were carried out by MALDI-Biotyper software; the protein fingerprint library was built by using WHONET5.6 software was used to statistically analyze the drug sensitivity results, SPSS23.0 software was used to analyze the relationship between fingerprint typing and drug sensitivity, and the χ² test was used for intergroup comparisons. Results: A total of 1 841 strains of E. coli were detected in 4 582 positive blood culture specimens from January 2016 to December 2022, with a detection rate of 40.17%; the resistance rate of E. coli from blood sources to piperacillin/tazobactam and ceftriaxone was on the rise, and it was slightly decreased to cefepime, amikacin, levofloxacin, and sulfamethoxazole, and there was not much change to the rest of the drugs; After MALDI-Biotyper clustering analysis, the 1841 E. coli strains from Isolated From Blood Culture were classified into two major clusters and five subtypes, of which type Ⅰa1 accounted for about 40%, type Ⅰa2 accounted for about 2.7%, type Ⅰb accounted for about 3.8, type Ⅱa accounted for about 46%, and type Ⅱb accounted for about 7.5%. The detection rate of type Ⅰa1 E. coli was higher in general surgery (50.45%) and emergency surgery (50.92%), and the detection rate of type Ⅰb E. coli was higher in emergency medicine(10.05%)than in other departments. The drug sensitivity results of different subtypes were compared with each other, the resistance rate of type Ⅰa1 E. coli to cefepime was 21.3% higher than that of the remaining four types, and the difference was statistically significant (χ²=37.74,P=0.000); the resistance rate of type Ⅱ E. coli(>60%) to sulfamethoxazole was higher than that of type Ⅰ (<60%) as a whole, and the difference was statistically significant (χ²=15.248,P=0.004); and a preliminary database of homologous protein fingerprints of E. coli has been established E. coli homologous protein fingerprint library and validated. The drug susceptibility results of 1 288 E. coli strains in the validation set were statistically analyzed and compared with those in the training set. There was no significant difference(P>0.05). Conclusion: In recent years, the resistance rate of E. coli isolated from a hospital in Qinghai province to piperacillin/Tazobactam, cefepime, amicacin and other antibiotics has changed greatly. A fingerprint database of E. coli homologous protein was established, and it was found that the drug sensitivity data of E. coli were different among different fingerprint types. According to drug sensitivity, drug use could assist clinical experience and provide evidence for prevention and control of hospital illness.
Humans ; Blood Culture ; Escherichia coli ; Cefepime ; Retrospective Studies ; Drug Resistance ; Sulfamethoxazole ; Piperacillin ; Tazobactam

Objective: To explore the drug resistance of Isolated From Blood Culture Escherichia coli (E. coli) in a hospital in Qinghai over the past seven years, to evaluate the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the homologous origin of E. coli, and to establish a protein fingerprint library to match with it, adjuvant clinical experience medication so as to provide the basis for the prevention and control of hospital-acquired infections. Methods: Retrospective analysis of blood cultures sent to hospitals from January 2016 to December 2022. Drug resistance and resistance changes in E. coli.A total of 1 841 E. coli strains were isolated from Qinghai Provincial People's Hospital from January 2016 to December 2022; all strains were identified by MALDI-TOF MS, and the VITEK2.0 drug sensitivity analyzer was applied for drug sensitivity analysis of the strains, and the mass spectrometry homology analysis and self-constructed protein fingerprint library were carried out by MALDI-Biotyper software; the protein fingerprint library was built by using WHONET5.6 software was used to statistically analyze the drug sensitivity results, SPSS23.0 software was used to analyze the relationship between fingerprint typing and drug sensitivity, and the χ² test was used for intergroup comparisons. Results: A total of 1 841 strains of E. coli were detected in 4 582 positive blood culture specimens from January 2016 to December 2022, with a detection rate of 40.17%; the resistance rate of E. coli from blood sources to piperacillin/tazobactam and ceftriaxone was on the rise, and it was slightly decreased to cefepime, amikacin, levofloxacin, and sulfamethoxazole, and there was not much change to the rest of the drugs; After MALDI-Biotyper clustering analysis, the 1841 E. coli strains from Isolated From Blood Culture were classified into two major clusters and five subtypes, of which type Ⅰa1 accounted for about 40%, type Ⅰa2 accounted for about 2.7%, type Ⅰb accounted for about 3.8, type Ⅱa accounted for about 46%, and type Ⅱb accounted for about 7.5%. The detection rate of type Ⅰa1 E. coli was higher in general surgery (50.45%) and emergency surgery (50.92%), and the detection rate of type Ⅰb E. coli was higher in emergency medicine(10.05%)than in other departments. The drug sensitivity results of different subtypes were compared with each other, the resistance rate of type Ⅰa1 E. coli to cefepime was 21.3% higher than that of the remaining four types, and the difference was statistically significant (χ²=37.74,P=0.000); the resistance rate of type Ⅱ E. coli(>60%) to sulfamethoxazole was higher than that of type Ⅰ (<60%) as a whole, and the difference was statistically significant (χ²=15.248,P=0.004); and a preliminary database of homologous protein fingerprints of E. coli has been established E. coli homologous protein fingerprint library and validated. The drug susceptibility results of 1 288 E. coli strains in the validation set were statistically analyzed and compared with those in the training set. There was no significant difference(P>0.05). Conclusion: In recent years, the resistance rate of E. coli isolated from a hospital in Qinghai province to piperacillin/Tazobactam, cefepime, amicacin and other antibiotics has changed greatly. A fingerprint database of E. coli homologous protein was established, and it was found that the drug sensitivity data of E. coli were different among different fingerprint types. According to drug sensitivity, drug use could assist clinical experience and provide evidence for prevention and control of hospital illness.
Humans ; Blood Culture ; Escherichia coli ; Cefepime ; Retrospective Studies ; Drug Resistance ; Sulfamethoxazole ; Piperacillin ; Tazobactam

ObjectiveTo investigate the mechanism of Ershiwuwei Guijiu pill in preventing and treating postmenopausal osteoporosis （PMOP） by activating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway and inhibiting excessive autophagy. MethodFemale SD rats were ovariectomized and randomly divided into the sham operation group （Sham）， the operation group （OVX）， the Ershiwuwei Guijiu pill （GJ） group， and the raloxifene hydrochloride （RLX） group， with 10 rats in each group. Enzyme-linked immunosorbent assay （ELISA） and colorimetric methods were used to detect the levels of estrogen， bone metabolism markers in serum， and total superoxide dismutase （T-SOD）， malondialdehyde （MDA）， and glutathione peroxidase （GSH-Px） in tibial tissue. Flow cytometry was used to detect reactive oxygen species （ROS） levels in bone marrow mesenchymal stem cells. Masson staining was used to observe pathological changes in the proximal tibia， and micro-computed tomography （Micro-CT） was used to observe changes in tibial microstructural parameters. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the expression of autophagy-related proteins Beclin1， microtubule-associated protein 1A/1B-light chain 3 （LC3）， autophagy-related 5 （Atg5）， as well as PI3K， Akt， and mTOR in tibial tissue. ResultCompared with the Sham group， the OVX group showed a significant decrease in serum levels of estradiol （E₂） and calcium ion （Ca²⁺）， and T-SOD， GSH-Px， PI3K， Akt， and mTOR mRNA levels in bone tissue （P<0.05， P<0.01）， significantly reduced bone mineral density （BMD）， bone surface/bone volume （BS/BV）， trabecular thickness （Tb.Th）， trabecular number （Tb.N）， and trabecular connectivity （Con） in the tibia （P<0.05， P<0.01）， thinner epiphyseal growth plate， and the bone marrow cavity filled with fat vacuoles. Moreover， the levels of phosphorus （P）， MDA， ROS， and mRNA and protein expression of Beclin1， LC3， and Atg5， as well as trabecular separation （Tb.Sp） were significantly elevated （P<0.05， P<0.01）. Compared with the OVX group， the GJ and RLX groups showed significant increases in serum E₂ and Ca²⁺， and bone tissue levels of SOD， GSH-Px， and the mRNA levels of PI3K， Akt， and mTOR （P<0.05， P<0.01）， significantly increased BMD， BS/BV， Tb.Th， Tb.N， and Con in the tibia， thickened epiphyseal growth plate， and significantly reduced fat vacuoles in the bone marrow cavity （P<0.05， P<0.01）. Additionally， the levels of P， MDA， ROS， Beclin1， LC3， Atg5 mRNA and proteins， and Tb.Sp were significantly decreased （P<0.05， P<0.01）. The ratios of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR， which were significantly reduced in the OVX group （P<0.01）， were significantly increased in the GJ and RLX groups （P<0.01）. ConclusionThe Ershiwuwei Guijiu pill reduces oxidative stress and inhibits autophagy， thereby preventing and treating postmenopausal osteoporosis. Its mechanism may be related to the activation of the PI3K/Akt/mTOR signaling pathway， which inhibits autophagy.

Objective To investigate the postoperative complications of ex vivo liver resection combined with autologous liver transplantation in the treatment of end-stage hepatic alveolar echinococcosis at high altitude and related prevention and treatment strategies. Methods Surgical data and follow-up data were collected from 11 patients with end-stage hepatic alveolar echinococcosis who underwent autologous liver transplantation in Qinghai People's Hospital from January 2013 to March 2019, and intraoperative and postoperative conditions were analyzed. Results All 11 patients underwent autologous liver transplantation successfully, without intraoperative death, among whom 2(18.18%) underwent hemi-extracorporeal hepatectomy and 9 (81.82%) underwent total extracorporeal hepatectomy. For the reconstruction of the retrohepatic inferior vena cava, 2 patients (18.18%) underwent reconstruction with the autologous great saphenous vein, 4 patients (36.36%) underwent reconstruction with artificial vessels, and the autologous retrohepatic inferior vena cava was preserved in 5 patients (45.45%). For biliary reconstruction, 8 patients (72.73%) underwent choledochoenterostomy and 3 (27.27%) underwent choledochocholedochostomy. The main postoperative complications of the 11 patients included bleeding in 2 patients (18.18%), bile leakage and abdominal infection in 4 patients (36.36%), bilioenteric anastomotic stenosis in 1 patient (9.09%), thrombus in 2 patients (18.18%), pulmonary infection and pleural effusion in 2 patients (18.18%), and echinococcosis recurrence in 1 patient (9.09%). Of all 11 patients, 2 (18.18%) died during the perioperative period, and the other 9 patients (81.82%) were improved and discharged. Conclusion Bleeding, biliary complications, and infection are the main causes of death in patients undergoing autologous liver transplantation at high altitude. An accurate understanding of surgical indication, careful multidisciplinary evaluation before surgery, superb operation during surgery, standardized surgical procedures, and fine perioperative management are the key to reducing perioperative mortality, avoiding and reducing postoperative complications, and achieving good long-term survival in patients undergoing autologous liver transplantation.

Objective:To observe the clinical efficacy of addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang to postmenopausal osteoporosis （PMO） with deficiency of spleen and kidney， and to investigate its regulation effect on immune inflammatory factors. Method:One hundred and sixty patients were randomly divided into observation group and control group， with 80 cases in each group. Both groups got comprehensive western medicine treatment measures. Patients in control group additionally got Zhuanggu Zhitong capsule， 4 capsules/time， 3 times/day. Patients in observation group additionally got addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang， 1 dose/day. The treatment was continued for 24 weeks. Before and after treatment， lumbar L2-4 bone mineral density （BMD） was detected by Dual energy X-ray absorptiometry （DXA） and lumbar BMD was detected by quantitative CT （QCT）. Scores of traditional Chinese medicine（TCM） syndromes and Chinese osteoporosis-targeted quality of life questionnaire （COQOL） were graded. Levels of Estradiol （E₂）， type Ⅰ procollagen amino terminal pro peptide （PINP）， serum osteocalcin （OC）， osteoprotegerin （OPG）， type Ⅰ collagen cross-linked C-terminal peptide （S-CTX）， tartrate resistant acid phosphatase （TRACP） and urinary pyridinoline （PYD） were detected. Levels of CD4⁺ T cells， CD8⁺ T cells， interleukin-17 （IL-17）， tumor necrosis factor-α （TNF-α）， γ-interferon（IFN-γ） and interleukin-4 （IL-4） were calculated. The proportion of T helper cell （Th）17 and regulatory T cell （Treg） in CD4⁺ T cells was calculated. Besides， the safety was evaluated. Result:Bone density was detected by DXA in observation group， and its T-value and bone density detected by QCT were all higher than those in control group （P<0.01）. After treatment， scores of TCM syndrome and COQOL were lower than those in control group （P<0.01）. Levels of PINP， OC， S-CTX， TRACP and PYD/Cr were all lower than those in control group （P<0.01）. Levels of OPG， CD8⁺ and Treg were higher than those in control group （P<0.05）， levels of Th17， Th17/Treg， CD4⁺/CD8⁺， IL-17， TNF-α and IFN-γ were lower （P<0.01）， and levels of IL-4 and E₂ were higher than those in control group （P<0.01）. The clinical efficacy in observation group was better than that in control group （Z=2.103， P<0.05）. Conclusion:On the basis of calcium and vitamin D supplementation， Jinkui Shenqiwan combined with Buzhong Yiqitang can improve levels of E₂ and bone density， reduce clinical symptoms， improve quality of life， regulate bone metabolism index and immune inflammation reaction， with better clinical efficacy and safety.

OBJECTIVE: To explore the relationship between metabolic syndrome( MS) and occupational stress among workers in high altitude area. METHODS: A cross-sectional study was conducted in 940 workers as research subjects who were engaged in petroleum smelting,thermal power generation and financial management et al. in high altitude area. A questionnaire survey was conducted using the Occupational Stress Inventory-Revised for occupational stress, The occupational health examination was performed to measure waist circumference,blood pressure,blood sugar and blood lipids. RESULTS: The abnormal rates in obesity,triglycerides,high-density lipoprotein,blood pressure and fasting blood glucose were 28. 3%,16. 3%,14. 3%,13. 3% and 6. 5%, respectively. The detection rate of MS was 5. 0%( 47/940). The multivariate logistic regression analysis showed that the risk of elevated MS decreased with the increased of occupational stress level( P < 0. 05),and the risk of elevated MS increased with the increased of occupational stress reaction severity( P < 0. 05),after adjusting the confounding factors of gender and labor nature. CONCLUSION: The higher occupational stress reaction of the workers in high altitude area,the higher the risk of MS.

OBJECTIVE: To comprehend the status of knowledge-attitude-practice( KAP) and its effect on blood lead in workers exposed to lead.METHODS: Three hundred and fifty-one first-line lead exposed workers in the smelting industry were chosen as study subjects by cluster sampling method.Blood lead levels in peripheral venous blood were detected.Questionnaire survey was conducted by self-compiled Questionnaire of Knowledge-attitude-practice on Occupational Health in Lead Workers.RESULTS: Among the study subjects,145 workers had abnormal blood lead level( ≥600 μg/L),the abnormal rate was 41.3%.The correct scoring rate in occupational knowledge,attitude,and practice were 25.1%,45.3% and 15.7%,respectively.Multivariate logistic regression analysis results showed that the four risk factors of high blood lead level were wearing no personal protective equipment,not bathing and changing clothes before returning home,not gargling and washing hands before meals,smoking and eating in workplace.CONCLUSION: Poor occupational behaviors can increase the risk of high blood lead level in lead exposed workers.